JP2018502309A - Apparatus and cartridge for performing an assay in a closed sample preparation and reaction system - Google Patents

Abstract

In one embodiment, a composite fluid processing cartridge includes a sample well, a deformable fluid chamber, a mixing well having a mixer disposed therein, a lysis chamber including a lysis mixer, and microdroplet manipulation. An electrowetting grid and an electronic sensor array configured to detect an analyte of interest. An apparatus for processing a cartridge is configured to receive a cartridge and selectively apply thermal energy, magnetic force, and electrical connections to one or more individual locations on the cartridge in a specific sequence ( It is further configured to compress the deformable chamber (s). [Selection] Figure 1

Description

  The subject matter of this disclosure relates to systems and methods for providing clinical and molecular diagnostics in an integrated composite device that provides sample-to-answer results. In particular, this disclosure provides a cartridge and a plurality of reagents to which a sample can be added, including reagents, buffers, and other processing materials for performing diagnostic assays or other processing on the sample. Of such cartridges are configured to independently process such cartridges.

  One major problem in the field of clinical and molecular diagnostics is the “sample-answer” system, which requires minimal sample handling and preparation and minimal requirements for experienced clinical laboratory personnel. It is the ability to have. Many systems have been proposed, but to date there are virtually no such commercial systems that meet these requirements well. Aspects of the invention provide such an integrated composite system.

  The present invention provides molecular diagnostic methods based on detection of analytes of interest, including nucleic acids, and composites. The systems described herein currently require some off-chip handling of the sample, typically including sample extraction (eg, cell lysis) and sample preparation prior to detection. In contrast to commercial systems, it is a fully integrated “sample-answer” system. Thus, according to current system aspects, the sample is mounted on a test platform and the analyte sample of interest is extracted and amplified as needed (eg, isothermal amplification can be used as well, but the subject If the analyte is a nucleic acid, the polymerase chain reaction technique is used) and then electrochemical detection is used herein, generally referred to as a “composite cartridge” or “fluid sample processing cartridge”. Everything on the microfluidic platform is detected.

  The unique utility of this system is the simplicity and high speed of this integrated system. For example, before introducing a sample into the system, only two operations are required, making it easy to use and not requiring highly trained laboratory personnel. An important advantage to the system is also that in some embodiments, generally only about 45-90 minutes from sample introduction to reporting of assay results, most results are reported in about 60-70 minutes or less. Is the speed from sample to answer. This represents a major advantage for both laboratories and physicians who rely on rapid analysis for diagnosis and rapid initiation of appropriate treatment. In addition, as outlined below, the ability to analyze multiple cartridges with fully random access, as well as the ability to perform highly complex multiple tests on a single cartridge, This is a great advantage in the context of clinical laboratories. A further advantage of the system is that it can be used for point-of-care diagnosis.

  Accordingly, aspects of the present invention are directed to an integrated system that allows the analyte of interest to be detected from a sample.

  For example, aspects of the present invention may include a substrate, a sample well formed in the substrate, a closure, a deformable fluid chamber supported on the substrate, a mixing well formed in the substrate, and a mixing well. And a fluid sample processing cartridge comprising a driven mixing device. The sample well is configured to receive a volume of fluid sample and the closure is configured to be selectively placed over the sample well. The deformable fluid chamber is configured to rupture to expel at least a portion of the fluid from the fluid chamber when an external compressive force is applied, so as to retain the fluid in the undeformed state. Is done. The deformable fluid chamber is in fluid communication with the sample well through a channel formed in the substrate. The mixing well passes through the sample well and fluid through a channel formed in the substrate, extends from a first peripheral wall defining the well, a first floor, and a channel that communicates the mixing well to the sample well. A fluid inlet snorkel that extends on one side of the peripheral wall and stops under the upper end of the first peripheral wall. The drive mixing device is configured and arranged to mix the contents of the mixing well.

  According to a further aspect of the invention, the fluid inlet snorkel extends on the outer surface of the first peripheral wall and stops at the opening formed in the first peripheral wall.

  According to a further aspect of the present invention, the sample well extends from one end of the second peripheral wall to the second peripheral wall and second floor defining the well and below the upper end of the second peripheral wall. A fluid inlet snorkel that stops.

  According to a further aspect of the invention, the mixing well further comprises an exit port comprising one or more openings formed in the floor of the mix well, the floor being tapered downward toward the exit port. ing.

  According to a further aspect of the invention, the drive mixing device can be driven by a first impeller arranged for rotation in the mixing well and a mating gear of the device into which the fluid sample processing cartridge is inserted. And a gear configured to rotate the first impeller when meshed with the meshing gear.

  According to a further aspect of the invention, the sample processing cartridge comprises a lysis chamber comprising a plurality of lysis beads, wherein the lysis chamber is disposed along a channel formed in the substrate and connecting the mixing well and the sample well. In addition, fluid flowing from the sample well to the mixing well flows through the lysis chamber. The sample processing cartridge further comprises a bead mixer disposed at least partially within the lysis chamber and configured to agitate the lysis beads and fluid flowing through the lysis chamber.

  According to a further aspect of the invention, the sample processing cartridge comprises a first optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the sample well, and a first comprising an enlarged portion of the channel connecting the lysis chamber to the mixing well. And two optical interfaces.

  According to a further aspect of the invention, the bead mixer comprises a motor mounted in the substrate and a second impeller disposed in the dissolution chamber and mounted on the output shaft of the motor.

  According to a further aspect of the invention, the lysis chamber includes a fluid inlet and a fluid outlet and is disposed on each of the fluid inlet and the fluid outlet and configured to hold a lysis bead in the lysis chamber. A mesh filter is further provided.

  According to a further aspect of the present invention, a sample processing cartridge is formed in the substrate, the pressure port configured to couple the substrate to an external fluid pressure source, and the substrate connecting the pressure port to the sample well. A channel.

  According to a further aspect of the invention, the sample processing cartridge comprises a waste chamber formed in the substrate, the waste chamber passing the mixing well and the fluid through the channel formed in the substrate, and the channel formed in the substrate. A fluid exit port formed in the substrate through which the mixing well and fluid are routed, and disposed in the substrate to selectively flow or stop the flow of fluid from the mixing well to the waste chamber A first externally actuable control valve configured and arranged and arranged in the substrate and configured to selectively enable or stop flow of fluid from the mixing well to the fluid ejection port; And a second externally operable control valve disposed.

  According to a further aspect of the invention, the sample processing cartridge further comprises a capture chamber disposed along a channel connecting the mixing well and the waste chamber.

  According to a further aspect of the present invention, a sample processing cartridge comprises a passive valve assembly disposed in a substrate, and a pressure valve formed in the substrate and formed in the substrate with a pressure port through which the passive valve assembly and pressure are passed. In addition. The passive valve assembly closes when the pressure in the mixing well is not greater than the threshold pressure to prevent fluid from flowing from the mixing well and opens when the pressure in the mixing well rises above the threshold pressure. , Configured and arranged to allow fluid to flow from the mixing well. When the pressure port is closed, the pressure in the mixing well reaches a threshold pressure that opens the passive valve assembly, allowing fluid to flow from the mixing well, and when the pressure port is open, The pressure does not reach the threshold pressure and the passive valve assembly is closed.

  According to a further aspect of the invention, the sample processing cartridge further comprises a lance blister associated with the deformable fluid chamber. The lance blister includes beads that are connected to an associated deformable fluid chamber or held in the lance blister by a connectable and breakable septum. The lance blister is configured to rupture when an external compressive force is applied, thereby pushing the beads through the breakable septum.

  According to a further aspect of the invention, the sample processing cartridge further comprises an external burr that wraps at least a portion of the cartridge from the outside.

  According to a further aspect of the invention, the sample processing cartridge further comprises a plurality of deformable fluid chambers, each of which is a lysis buffer, a wash buffer, an oil, a rehydration buffer, a target capture bead. And one or more substances selected from the group consisting of binding buffers.

  According to a further aspect of the present invention, a sample processing cartridge is formed on the substrate, the first fluid ejection port passing the mixing well and fluid through the channel formed in the substrate, and the second formed in the substrate. The apparatus further comprises a fluid exit port and at least two deformable fluid chambers. One of the two deformable fluid chambers passes the mixing well and fluid through a channel formed in the substrate, the other of the two deformable fluid chambers has the mixing well and the first fluid ejection port. The second fluid exit port and the fluid are passed through a channel formed in the substrate, which is different from the communicating channel.

  According to a further aspect of the invention, the deformable fluid chamber through the mixing well and the fluid includes a lysis buffer, a wash buffer, a target capture bead, or a binding buffer, and a second fluid ejection and fluid. The deformable fluid chamber leading to it contains oil or a rehydration buffer.

  A further aspect of the invention is embodied by a fluid sample processing cartridge comprising a sample preparation module comprising a reaction module. The sample preparation module includes a substrate, a sample well formed on the substrate and configured to receive a volume of fluid sample, a closure configured to be selectively placed on top of the sample well, And is configured to collapse when an external compressive force is applied to retain the fluid therein and expel at least a portion of the fluid from the first fluid chamber. A first deformable fluid chamber, the first deformable fluid chamber being adapted to pass the sample well and fluid through a channel formed in the substrate, and a channel formed in the substrate. A mixing well adapted to allow fluid to flow through the sample well and disposed within the mixing well and configured to mix the contents of the mixing well A drive mixing device disposed; and a first fluid outlet port formed in the substrate, the first fluid outlet port adapted to pass fluid through the mixing well through a channel formed in the substrate; . The reaction module is attached to the sample preparation module and is configured to receive fluid from the sample preparation module via a fluid outlet port formed in the sample preparation module. The reaction module has an upper plate with an upper surface, and at least partially surrounds the periphery of the upper surface, is in contact with the surface of the sample preparation module and seals the fluid, and an intervening space between the upper surface and the surface of the sample preparation module A rising wall that forms a fluid, a sample chamber fluidly coupled to the first fluid outlet port of the sample preparation module, a reagent chamber, a detection chamber, a lower surface of the upper plate, and a fluid processing panel and upper plate And a fluid processing panel that defines a reaction / processing space. The reaction / processing space may be open or open to the sample chamber, reaction chamber, and detection chamber.

  According to a further aspect of the invention, the reaction module includes an inlet port through which the fluid sample enters the sample chamber, and includes a gap between the first fluid exit port of the sample preparation module and the inlet port of the sample chamber; The gap is open to the intervening space.

  According to a further aspect of the invention, the first fluid exit port of the sample preparation module comprises an outlet channel formed through the frustoconical nipple.

  According to a further aspect of the invention, the reaction module of the fluid sample processing cartridge further comprises an electronic sensor array disposed in each detection chamber.

  According to a further aspect of the present invention, the top plate of the reaction module further comprises one or more bubble traps, each bubble trap being open to the reaction and processing space and to the intervening space. An open vent opening.

  According to a further aspect of the invention, the sample preparation module is supported on the substrate and retains the fluid therein when undeformed and ruptures when an external compressive force is applied. The apparatus further comprises a second deformable fluid chamber configured to discharge at least a portion of the fluid from the chamber, and a second fluid ejection port formed in the substrate. The second fluid exit port is channeled to a second deformable fluid chamber and fluid through a channel formed in the substrate, and the reaction and processing space is hydrodynamically connected to the second fluid exit port of the sample preparation module. Combined with

  According to a further aspect of the invention, the mixing well extends around the perimeter wall and floor defining the well and one side of the perimeter wall extending from the channel leading to the mixing well to the sample well and stops below the upper end of the perimeter wall. A fluid inlet snorkel.

  According to a further aspect of the invention, the fluid inlet snorkel extends on the outer surface of the peripheral wall and stops at an opening formed in the peripheral wall.

  According to a further aspect of the invention, the mixing well further comprises an exit port comprising one or more openings formed in the floor of the mix well, the floor being tapered downward toward the exit port. Yes.

  According to a further aspect of the invention, the drive mixing device is meshed so that it can be driven by a first impeller rotatably disposed within the mixing well and a meshing gear of the device into which the fluid sample processing cartridge is inserted. And a gear configured to rotate the first impeller when the meshing gear meshes.

  According to a further aspect of the invention, the sample preparation module further comprises a lysis chamber comprising a plurality of lysis beads and formed along a channel formed in the substrate and connecting the mixing well and the sample well. The fluid flowing from the sample well to the mixing well flows through the lysis chamber, and the sample conditioning module is disposed at least partially within the lysis chamber to agitate the lysis beads and the fluid flowing through the lysis chamber. A constructed and arranged bead mixer is provided.

  According to a further aspect of the invention, the bead mixer comprises a motor mounted in the substrate and a second impeller disposed in the dissolution chamber and mounted on the output shaft of the motor.

  According to a further aspect of the invention, the fluid sample processing cartridge comprises a first optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the sample well and an enlarged portion of the channel connecting the lysis chamber to the mixing well. And a second optical interface.

  According to a further aspect of the invention, the lysis chamber includes a fluid inlet and a fluid outlet, and is disposed on each fluid inlet and fluid outlet, and includes a mesh filter configured to hold lysis beads in the lysis chamber. In addition.

  According to a further aspect of the invention, the sample preparation module is formed in the substrate and formed in the substrate configured to couple the substrate to an external fluid pressure source and the pressure port connected to the sample well. A further channel.

  According to a further aspect of the present invention, the sample preparation module is formed in the substrate and disposed in the substrate through the channel formed in the substrate and through the mixing well and fluid, and disposed in the substrate, from the mixing well to the disposal chamber. A first externally actuable control valve configured and arranged to selectively flow fluid or to prevent fluid flow and a substrate to selectively flow fluid from the mixing well to the exit port; And a second externally operable control valve configured and arranged to prevent fluid flow.

  According to a further aspect of the invention, the sample preparation module further comprises a capture chamber disposed along a channel connecting the mixing well and the waste chamber.

  According to a further aspect of the invention, the sample preparation module is disposed in the substrate and is closed when the pressure in the mixing well is not greater than the threshold pressure, preventing fluid from flowing from the mixing well, A passive valve assembly configured and arranged to open and allow fluid to flow from the mixing well when the pressure inside rises above a threshold pressure, and a pressure conduit formed in the substrate and formed in the substrate And further includes a passive valve assembly and a pressure port through which pressure is passed. When the pressure port is closed, the pressure in the mixing well reaches a threshold pressure that opens the passive valve assembly, allowing fluid to flow from the mixing well, and when the pressure port is open, in the mixing well Pressure does not reach the threshold pressure and the passive valve assembly is closed.

  According to a further aspect of the invention, the sample preparation module further comprises a lance blister associated with the deformable fluid chamber. The lance blister includes beads that are connected to an associated deformable fluid chamber or held in the lance blister by a connectable and breakable septum. The lance blister is configured to rupture when an external compressive force is applied, thereby pushing the beads through the breakable septum.

  According to a further aspect of the invention, the external flash wraps at least a portion of the cartridge from the outside.

  According to a further aspect of the invention, the sample preparation module further comprises a plurality of deformable fluid chambers, each fluid chamber comprising a lysis buffer, a wash buffer, an oil, a rehydration buffer, a target capture bead, And a substance selected from the group consisting of binding buffers.

  A further aspect of the invention is to support a fluid on a flat substrate and retain fluid in its interior in an undeformed state, rupture and apply fluid from the fluid chamber when an external compressive force is applied. And a device configured to process a fluid sample processing cartridge including a deformable fluid chamber configured to discharge at least a portion of the fluid chamber. The apparatus includes a cartridge carriage assembly configured to receive and hold a fluid sample processing cartridge inserted into the apparatus. The heating and control assembly is disposed adjacent to the cartridge carriage assembly and is in operative contact with the cartridge carried within the cartridge carriage assembly and a first position not in operative contact with the cartridge carried within the cartridge carriage assembly. Configured for movement relative to the cartridge carriage assembly between a second position. Each of the one or more movable magnet assemblies includes a heating and control assembly between a first position that does not substantially apply a magnetic force to the cartridge and a second position that applies a magnetic force to a corresponding individual portion of the cartridge. It is mounted for movement relative to the cartridge, independent of the cartridge. The cam block assembly is configured for powered movement, wherein the cam block assembly is powered between a first position of the heating and control assembly and a second position of the heating and control assembly. Operatively coupled to the heating and control assembly for conversion to movement of the heating and control assembly relative to the carriage assembly. The cam block assembly translates the powered movement of the cam block assembly into movement of each magnet assembly relative to the cartridge carriage assembly between the first position of the magnet assembly and the second position of the magnet assembly. Operably coupled to one or more movable magnet assemblies. The deformable chamber compression assembly is configured to selectively apply an external compressive force to the deformable fluid chamber to rupture the deformable chamber and discharge at least a portion of the fluid from the fluid chamber. .

  According to a further aspect of the invention, the heating and control assembly includes one or more heaters configured to apply a thermal gradient to a corresponding individual portion of the cartridge when the heating and control assembly is in the second position. And a connector board including one or more electrical connector elements configured to provide an electrical connection between the device and the cartridge when the heating and control assembly is in the second position.

  According to a further aspect of the invention, the deformable chamber compression assembly includes a cam follower plate configured for powered movement in a first direction substantially parallel to a plane of the substrate and a deformable cartridge. A compression mechanism configured to apply a force to compress the chamber against the substrate by movement in a second direction having a component substantially perpendicular to the plane of the substrate in relation to the chamber. The cam follower plate translates the movement of the cam follower plate in the first direction into movement of the compression mechanism in the second direction, thereby operably operating the compression mechanism to apply an external compression force to the chamber. Combined.

  According to a further aspect of the invention, the apparatus further comprises an air pump and an air port connected to the air pump, the air port being connected to the pressure port of the fluid sample processing cartridge when the cartridge is inserted into the apparatus. Is configured to be coupled to.

  According to a further aspect of the invention, the apparatus further comprises a photodetector configured to detect fluid flowing through a portion of the fluid sample processing cartridge.

  According to a further aspect of the invention, the fluid sample processing cartridge includes a drive mixing device that includes a drive gear, and the device further includes a mixing motor assembly that includes a powered drive gear. The mixing motor moves between a first position where the drive gear is not meshed with the drive gear of the drive mixing device and a second position where the drive gear is operatively meshed with the drive gear to operate the drive mixing device. Is possible. The cam block assembly converts the powered movement of the cam block assembly into a movement of the mixing motor assembly between a first position of the mixing motor assembly and a second position of the mixing motor assembly. Operatively coupled to the assembly.

  According to a further aspect of the invention, the apparatus further comprises a heater cooling assembly comprising a fan and a cooling duct configured to direct an air flow from the fan to a portion of one of the heater assemblies.

  According to a further aspect of the present invention, the cartridge carriage assembly includes a cartridge holder configured to hold a cartridge to be inserted and a cartridge holder biased to a cartridge latch position to hold the cartridge within the cartridge holder. A cartridge latch configured to latch a cartridge inserted into the cartridge and configured to automatically push the cartridge away from the cartridge holder when the cartridge latch is released from the cartridge latch position. A cartridge discharge mechanism.

  According to a further aspect of the invention, the heating and control assembly comprises a support plate on which one or more heater assemblies and a connector board are supported. The support plate is mounted in a constraining structure that prevents horizontal movement of the support plate but allows vertical movement of the support plate to allow movement of the heating and control assembly between the first and second positions. The

  According to a further aspect of the invention, the heater assembly of the heating and control assembly includes a resistance heating element attached to the connector board, and a heat spreader comprising a thermally conductive material thermally coupled to the resistance heating element. Prepare.

  According to a further aspect of the invention, one of the heater assemblies of the heating and control assembly includes a thermoelectric element, a thermal diffuser comprising a thermally conductive material thermally coupled to the thermoelectric element, a thermoelectric element and a plurality of heats. A heat sink including a panel in thermal contact with the dissipative rod.

  According to a further aspect of the invention, the electrical connector element of the connector board of the heating and control assembly comprises a plurality of connector pin arrays, each connector pin array comprising a plurality of pogo pins.

  According to a further aspect of the invention, one of the movable magnet assemblies is mounted on the spindle such that it can rotate about the spindle between a first position and a second position of the magnet assembly. A magnet holder, a magnet supported on the magnet holder, an actuator bracket extending from the magnet holder, and a torsion spring configured to bias the magnet holder to a rotational position corresponding to the first position of the magnet assembly; .

  According to a further aspect of the invention, one of the movable magnet assemblies is mounted on the spindle so as to be rotatable about the spindle between a first position and a second position of the magnet assembly. A magnet holder frame, a magnet array disposed in the magnet holder frame, a converging magnet disposed in an opening formed in the magnet holder frame and configured to converge the magnetic force of the magnet array, and a magnet holder frame An extending actuator bracket and a torsion spring configured to bias the magnet holder frame to a rotational position corresponding to a first position of the magnet assembly.

  According to a further aspect of the invention, the cam block assembly is moved each by a magnetic actuator coupled in part to the cam block assembly such that the cam block assembly is movable by a powered movement of the cam block assembly. Each magnet assembly is operably coupled to a possible magnet assembly such that a magnet actuator moves with the cam block assembly to cause a corresponding rotation of the magnet assembly from a first position to a second position. Including a tab configured to engage with the actuator bracket.

  According to a further aspect of the present invention, a cam block assembly is attached to a cam frame, a cam block motor coupled to the cam frame and configured to provide powered movement of the cam frame, and the cam frame. First and second cam rails. Each cam rail has two cam slots. The cam block assembly is operably coupled to the heating and control assembly by a cam follower extending from the heating and control assembly to the cam slot, and movement of the cam frame and cam rail relative to the heating and control assembly is a first of the heating and control assembly. A cam follower and a cam slot for moving the cam follower between a respective first segment of the cam slot corresponding to the position and a respective second segment of the cam slot corresponding to the second position of the heating and control assembly; Cause the corresponding relative movement between.

  According to a further aspect of the present invention, the cam frame includes a first longitudinal circle extending along one side of the heating and control assembly and a second extending along opposite sides of the heating and control assembly. A long circular material and a cross circular material extending between first and second long circular materials. Each cam rail is attached to one of the first and second longitudinal members.

  According to a further aspect of the invention, the compression mechanism of the deformable chamber compression assembly has a cam surface so that it can pivot about one end of the cam arm and a compression pad disposed at the opposite end of the cam arm. A cam arm mounted, wherein the cam arm has a first position where the compression pad does not contact the associated deformable chamber, and the associated deformable for the compression pad to at least partially rupture the chamber. Pivotable between a second position for applying a compressive force to the chamber.

  According to a further aspect of the invention, the deformable chamber compression assembly further comprises a cam arm plate, wherein the cam arm of the compression mechanism is formed in the cam arm plate for pivoting movement of the cam arm relative to the cam arm plate. It is mounted so as to be pivotable in the slot. The cam surface of the cam arm protrudes from a slot on the surface of the cam arm plate. The cam follower plate is engaged by a cam follower element of the cam follower plate that engages a cam surface of the compression mechanism during movement of the cam follower plate relative to the cam arm plate to cause the cam arm to pivot from a first position to a second position. Operatively coupled to the compression mechanism.

  According to a further aspect of the present invention, the cam follower plate includes a cam groove for receiving the cam surface of the cam arm protruding on the surface of the cam arm plate, and the cam follower element pivots from the first position to the second position on the cam arm. For operation, the cam arm plate includes a follower ridge disposed in a cam groove that contacts the cam surface when the cam follower plate moves relative to the cam arm plate.

  According to a further aspect of the present invention, the apparatus further comprises a plurality of compression mechanisms, each comprising a cam arm plate and a cam arm that is pivotably mounted within a slot formed in the cam arm surface, the cam follower plate Comprises a plurality of cam grooves, each cam groove being associated with at least one of the compression mechanisms and for pivoting a cam arm of the associated compression mechanism from a first position to a second position. Each cam groove includes a follower ridge disposed in a cam groove that contacts a cam surface of an associated compression mechanism as the cam follower plate moves relative to the arm plate.

  According to a further aspect of the invention, the sample processing cartridge includes a plurality of deformable fluid chambers, and the deformable chamber compression assembly comprises a plurality of compression mechanisms. Each compression mechanism is associated with one of the deformable fluid chambers, and the cam follower plate is operably coupled with the compression mechanism, and the movement of the cam follower plate in the first direction is in a second direction of the compression mechanism. To an external compression force, thereby applying an external compressive force to each of the associated chambers in a specific sequence.

  According to a further aspect of the invention, the fluid sample processing cartridge allows fluid flow through the valve when not actuated externally, and fluid flow through the valve when actuated externally. Including an externally operable control valve configured to selectively control fluid flow. The apparatus is configured to operate the associated externally operable control valve by movement in a second direction having a component substantially perpendicular to the plane of the substrate, associated with the externally operable control valve of the sample processing cartridge. A valve actuator compression mechanism is further provided. The cam follower plate is operably coupled to the valve actuator compression mechanism to convert movement of the cam follower plate in a first direction to movement of the valve actuator compression mechanism in a second direction, thereby associated externally operable. Activate the control valve.

  Other features and characteristics of the subject matter of this disclosure, together with the method of operation, the function of the associated elements and components of the structure, and the economics of manufacture, are all included in the accompanying drawings, which form a part hereof. In view of the following description and any appended claims, reference will become more apparent.

  The accompanying drawings, which are incorporated herein and form a part of this specification, illustrate various embodiments of the subject matter of this disclosure. In the drawings, like reference numbers indicate identical or functionally similar elements.

FIG. 6 is a top perspective view of a composite cartridge embodying aspects of the present invention. It is an upper top view of a composite cartridge. It is an upper top view of the composite cartridge to which the identification label is attached. It is a disassembled perspective view of a composite cartridge. FIG. 6 is a partial perspective view of a cross section of a deformable fluid compartment (or blister) of a composite cartridge. 2 is a detailed perspective view of a sample well and a sample cap of a composite cartridge. FIG. FIG. 7 is a cross-sectional perspective view of the sample well taken along line 7-7 in FIG. FIG. 3 is a detailed perspective view of a mixing well and a mixer of a composite cartridge. FIG. 5 is a detailed perspective view of another mixing well of the composite cartridge. FIG. 8D is a top plan view of the mixing well of FIG. 8C. FIG. 9 is a cross-sectional view of the mixing well and mixer taken along line 9-9 in FIG. FIG. 9 is a cross-sectional view of another mixing well of FIGS. 8B and 8C and another mixer disposed therein. It is a detailed perspective view of the passive valve of a composite cartridge. FIG. 3 is a cross-sectional perspective view of the passive valve along line 11-11 in FIG. FIG. 13 is a cross-sectional perspective view of the dissolution chamber and bead mixer taken along line 12-12 in FIG. FIG. 13 is a cross-sectional perspective view of the active valve assembly taken along line 13-13 in FIG. FIG. 5 is a cross-sectional perspective view of an active valve that is activated by an external valve actuator. It is an upper top view of the sample preparation module of a composite cartridge. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. FIG. 4 shows a top plan view of a sample preparation module, each showing different steps of the sample preparation process performed within the module. It is a top perspective view of the upper plate of the reaction module of a composite cartridge. It is a lower perspective view of an upper plate. It is an upper top view of an upper plate. It is a lower part top view of an upper plate. FIG. 25 is a cross-sectional perspective view of the reaction module taken along line 28-28 in FIG. FIG. 25 is a cross-sectional perspective view of the reaction module taken along line 29-29 in FIG. 24. It is a detailed perspective view of the fluid inlet of the reaction module. It is a fragmentary sectional view in alignment with line 31-31 in FIG. 1 is a front view of an apparatus embodying aspects of the present invention. It is a front perspective view of the control console of an apparatus. It is a front perspective view of the processing module of an apparatus. FIG. 3 is a front perspective view of a processing module with one side wall of the module removed to show the internal components of the processing module. FIG. 6 is a rear perspective view of the processing module with one side wall and the back wall of the module removed to show the internal components of the processing module. It is the front perspective view of the processing module by which one side wall and one back wall of the module were removed, and one processing bay of the processing module was disassembled from the module. It is the front and right side perspective view of the processing bay which embody the aspect of this invention. It is a front and left side perspective view of a processing bay. It is a back surface of a processing bay, and a right perspective view. It is the front, right side, and exploded perspective view of a processing bay. 3 is an exploded perspective view of a cartridge processing assembly in a processing bay. FIG. FIG. 6 is an exploded perspective view of a heating and control assembly of the cartridge processing assembly. FIG. 6 is a top plan view of the connector PCB and magnet of the heating and control assembly of the cartridge processing assembly. FIG. 5 is an exploded perspective view of a detection Peltier heater assembly of the heating and control assembly. FIG. 6 is an exploded perspective view of a cartridge carriage assembly of the cartridge processing assembly. FIG. 5 is an exploded perspective view of a cam frame assembly of a cartridge processing assembly. FIG. 6 is a cross-sectional perspective view of a cam frame and magnet actuator of a cartridge processing assembly. FIG. 6 is a top perspective view of a sample preparation magnet assembly of a cartridge processing assembly. FIG. 6 is a top perspective view of a cartridge magnet assembly of a cartridge processing assembly. FIG. 6 is a perspective view of a mixing motor assembly of a cartridge processing assembly. It is a disassembled perspective view of a mixing motor assembly. 3 is an exploded perspective view of a blister compression mechanism assembly in a processing bay. FIG. It is a partial lower plan view of a cam arm plate showing a compression pad of an array of compression mechanisms. FIG. 6 is a top perspective view of an array compression mechanism isolated from a cam arm plate. FIG. 6 is a bottom perspective view of an array compression mechanism isolated from a cam arm plate. It is an exploded perspective view of a single fluid blister compression mechanism. It is a disassembled perspective view of a single lance blister compression mechanism. It is a disassembled perspective view of a single valve actuator compression mechanism. It is a lower top view of the cam follower plate of a blister compression mechanism assembly. It is a lower perspective view of a cam follower plate. It is a lower part top view of the fluid processing panel of a reaction module. It is a top plan view of a fluid treatment panel. 2 is a flowchart illustrating an exemplary process that can be performed in a fluid treatment panel.

  While aspects of the subject matter of the present disclosure may be embodied in various forms, the following description and the accompanying drawings merely disclose some of these forms as specific examples of the subject matter. It is only intended. Accordingly, the subject matter of this disclosure is not intended to be limited to the forms or embodiments so described and illustrated.

  Unless defined otherwise, all technical terms, notations, other technical terms, or terminology used herein are those commonly understood by one of ordinary skill in the art to which this disclosure belongs. Has the same meaning. All patents, applications, published applications, and other published documents referenced herein are incorporated by reference in their entirety. If the definitions set forth in this section are contrary to the definitions set forth in the patents, applications, published applications, and other published documents incorporated herein by reference, or otherwise inconsistent, The definitions set forth take precedence over the definitions incorporated herein by reference.

  As used herein, unless otherwise indicated or indicated otherwise, “a” or “an” means “at least one” or “one or more”.

  This description may use relative spatial and / or directional phrases to describe the position and / or orientation of a component, device, location, feature, or part thereof. Unless specifically stated or otherwise indicated by the context of the description, but not limited to, top, bottom, top, bottom, bottom, top, top, bottom, left, right, Such phrases including front, back, near, adjacent, between, horizontal, vertical, diagonal, longitudinal, lateral, radial, axial, etc. in the drawings Used for convenience to reference such components, apparatus, locations, features, or portions thereof, and is not intended to be limiting.

  Moreover, unless otherwise stated, any particular dimensions mentioned in this description are merely representations of exemplary implementations of devices embodying aspects of the invention and are not intended to be limiting.

<Related applications>
This application is related to US Patent Application No. 14 / 062,860 (US Patent Application Publication No. 2014-0322706) and US Patent Application No. 14 / 062,865 (US Patent Application Publication No. 2014-0194305). The disclosures of each of which are incorporated herein by reference.

[Introduction]
In general, the system includes two components: one is a composite cartridge in which the sample is loaded and the various reagents, buffers, and others to perform the desired assay or other procedure. Of processing materials. And the other is a processing device into which a cartridge is inserted to perform sample processing and final detection of the analyte of interest.

  In various embodiments, the microfluidic platform relies on the formation of microdroplets and the ability to independently transport, coalesce, mix and / or process droplets. In various embodiments, such microdrop operations are performed using electrical control of surface tension (ie, electrowetting). In general, a liquid sample is contained in a microfluidic device, known as a processing module, between two parallel plates. One plate, called a fluid treatment panel, contains drive electrodes etched on its surface, and the other plate is a single electrode that is etched or grounded or set to a reference potential. Including any of the continuous planar electrodes ("biplanar electrowetting"), a hydrophobic insulator envelops the electrodes, and an electric field is generated between the electrodes on the opposing plates. , Creating a surface tension gradient that causes the droplets that overlap the powered electrode to move toward the electrode.In some embodiments, the active electrowetting electrodes are adjacent and “coplanar” It can be coplanar with the adjacent ground reference electrode, called “coplanar electrowetting”. With placement and control, droplets can be transported by transporting them sequentially between adjacent electrodes, and patterned electrodes can be arranged in a two-dimensional array Allows droplets to be transported to any location covered by the array, the space surrounding the droplets can be filled with a gas such as air or an insoluble fluid such as oil, etc. Suitable in many embodiments of the present invention.

  As the droplets containing the analyte of interest move across the surface, they can capture reagents and buffers. For example, when a dry reagent is placed on a surface (generally described herein as a printed circuit board, but additional surfaces may be used as will be understood by those skilled in the art) , Droplets moving through the area will capture and decompose reagents used in biological processes such as PCR amplification. Furthermore, as described more fully below, the addition from the sample preparation module placed on the substrate is accompanied by sample preparation, eg, lysis, purification, degradation, etc., prior to transporting the sample to the microfluidic platform. Allows the specific addition of buffers and other reagents, such as wash buffers.

  Aspects of the invention also include the use of electrochemical detection of the analyte of interest. Suitable electrochemical detection systems include U.S. Pat.No. 4,887,455; U.S. Pat.No. 5,591,578; U.S. Pat.No. 5,705,348; U.S. Pat.No. 5,770,365; U.S. Pat.No. 5,807,701; U.S. Pat.No. 5,824,473; U.S. Patent 6,013,170; U.S. Patent 6,013,459; U.S. Patent No. 6,033,601; U.S. Patent No. 6,063,573; U.S. Patent No. 6,090,933; U.S. Patent No. 6,096,273; U.S. Patent No. 6,192,351; U.S. Patent No. 6,221,583; U.S. Patent No. 6,232,062; U.S. Patent No. 6,236,951; U.S. Patent No. 6,248,229; U.S. Pat. U.S. Patent No. 6,376,232; U.S. Patent No. 6,431,016; U.S. Patent No. 6,432,723; U.S. Patent No. 6,479,240; U.S. Patent No. 6,495,323; U.S. Patent No. 6,518,024; ; US Patent 6,600,026 U.S. Patent 6,602,400; U.S. Patent 6,627,412; U.S. Patent 6,642,046; U.S. Patent 6,655,010; U.S. Patent 6,686,150; U.S. Patent No. 6,740,518; U.S. Patent No. US Patent No. 6,824,669; US Patent No. 6,833,267; US Patent No. 6,875,619; US Patent No. 6,942,771; US Patent No. 6,951,759; US Patent No. 6,960,467; US Patent No. 6,977,151; US Patent No. U.S. Patent 7,045,285; U.S. Patent 7,056,669; U.S. Patent 7,087,148; U.S. Patent 7,090,804; U.S. Patent 7,125,668; U.S. Patent 7,160,678; U.S. Patent No. U.S. Patent 7,312,087; U.S. Patent 7,381,525; U.S. Patent 7,381,533; U.S. Patent 7,384,749; U.S. Patent 7,393,645; U.S. Patent 7,514,228; U.S. Patent 7,534,331; US Patent No. 7,566,534; US Special U.S. Patent No. 7,579,145; U.S. Patent No. 7,582,419; U.S. Patent No. 7,595,153; U.S. Patent No. 7,601,507; U.S. Patent No. 7,655,129; U.S. Patent No. 7,713,711; U.S. Patent No. 7,935,481; U.S. Patent No. 8,012,743; U.S. Patent No. 8,114,661 and U.S. Publication No. 2012/01 81 186, the disclosures of each of which are expressly incorporated herein by reference. Incorporated into.

  In various embodiments, processed analyte droplets are transported to a detection zone on a fluid processing panel and specifically captured on individual detection electrodes using the systems described in many of the above patents. The See in particular US Pat. No. 7,160,678, US Pat. No. 7,393,645, and US Pat. No. 7,935,481. This detection system relies on the use of a label probe (in the case of nucleic acids) that includes an electrochemically active label that allows the presence of the analyte of interest to be a positive signal and allows detection of pathogens, disease states, and the like.

[sample]
Aspects of the present invention provide systems and methods for the detection of analytes of interest in a sample for diagnosing diseases and infections caused by pathogens (eg, bacteria, viruses, fungi, etc.). As will be appreciated by those skilled in the art, sample solutions include but are not limited to bodily fluids (including but not limited to blood, urine, serum, plasma, cerebrospinal fluid, lymph, saliva, nasopharynx Contains samples, anal / vaginal secretions, feces, tissue samples containing tissues suspected of containing cancer cells, virtually any organism sweat / semen, preferably mammalian samples, human samples Environmental samples (including but not limited to air, agriculture, water and soil samples, environmental specimens, and other collection kits); biological weapons samples; food and beverage samples, research samples (ie In the case of nucleic acids, the sample can generally be the product of an amplification reaction that includes both subject and signal amplification, such as a PCR amplification reaction, as described in WO / 1999/037819. This disclosure is incorporated herein by reference); can include any number of items including: purified samples of purified genetic DNA, RNA, proteins, etc .; raw samples (bacteria, viruses, genetic DNA, etc.); As will be appreciated by those skilled in the art, the sample could be subjected to virtually any experimental manipulation.

  The composite cartridge can be used to detect an analyte of interest in a patient sample. “Analyte of interest” or “analyte” or grammatical equivalents herein means any molecule or compound that is to be detected and binds to a binding species as defined below. be able to. Suitable analytes include, but are not limited to, environmental or clinical chemistry, including pesticides, insecticides, poisons, therapeutic agents and drugs of abuse, hormones, antibiotics, antibodies, organic materials, etc. Including but not limited to small chemical molecules such as contaminants or biomolecules. Suitable biomolecules include, but are not limited to, proteins (including enzymes, immunoglobulins and glycoproteins), nucleic acids, lipids, lectins, carbohydrates, hormones, whole cells (such as pathogen bacteria) prokaryotes and eukaryotes Organism cells, including mammalian tumor cells, etc.), viruses, spores and the like.

  In one embodiment, the analyte of interest is a protein (“protein of interest”). As will be appreciated by those skilled in the art, there are a great many possible proteinaceous analytes that can be detected using the present invention. “Protein” or grammatical equivalents refer to proteins, oligopeptides / peptides, derivatives / analogs that contain amino acids and amino acid analogs that do not occur in nature, and peptide-like structures. The side chains can be either (R) or (S) structures. In a preferred embodiment, the amino acid is (S) or L-structure. As discussed below, if a protein is used as a binding ligand, it may be preferable to utilize a protein analog to delay degradation due to contamination of the sample. Particularly preferred proteins of interest include enzymes; drugs; cells; antibodies; antigens, cell membrane antigens / receptors (nerves, hormonal, nutrients, cell surface receptors) or their ligands.

  In a preferred embodiment, the target analyte is a nucleic acid (“target nucleic acid”). The system can be used to diagnose diseases such as single nucleotide polymorphisms (SNPs) that cause disease (eg, cystic fibrosis) or exist in disease (eg, tumor mutation), as well as bacteria and viruses, etc. Patients can be used to diagnose specific pathogens that are exogenous.

  As will be appreciated by those skilled in the art, the present invention relies on both the nucleic acid of interest and other nucleic acid components such as capture probes and label probes used to detect the nucleic acid of interest. “Nucleic acid” or “oligonucleotide” or grammatical equivalents means at least two nucleotides covalently linked together. The nucleic acids of the invention may in some cases be included as primers or probes in which the nucleic acid analog can have another backbone, as outlined below, but in general phosphodiester bonds Would include. For example, phosphoramides (Beaucage et al., Tetrahedron 49 (10). T 925 (1993) and references; Letsinger, J. Org. Chem. 35: 3800 (1970); Sprinzl et al., Eur J. Biochem. 81: 579 (1 977); Letsinger et al., Nucl. Acids Res. 14: 3487 (1986); Sawai et al, Chem. Left. 805 (1984), Letsinger et al., J. Am. Chem. Soc. 1 10: 4470 (1988); and Pauwels et al., Chemica Scripta 26: 141 91986)), phosphorothioates (Mag et al., Nucleic Acids Res. 19: 1437 (1991); and US patents No. 5,644,048), dithiophosphoric acid (Briu et al, J. Am. Chem. Soc. 1 1 1: 2321 (1989), O-methylphophoroamidite linkages (see Eckstein, Oligonucleotides and Analogues : A Practical Approach, Oxford University Press), and peptide nucleic acid backbones and linkages (see Egholm, J. Am. Chem. Soc. 1 14: 1895 (1992); Meier et al., Chem. Int. Ed. Engl. 31: 1008 (1992); Nielsen, Nature, 365: 566 (1993); Carlsson et al., Natu re 380: 207 (1996)), all incorporated by reference, other similar nucleic acids include those with a positive backbone (Denpcy et al., Proc. Natl. Acad. Sci. USA 92: 6097 (1995). ); Having a bicycle structure containing a locked nucleic acid, Koshkin et al., J. Am. Chem. Soc. 120: 13252-3 (1998); nonionic backbone (US Pat. No. 5,386,023, US Pat. No. 5,637,684) No., U.S. Pat.No. 5,602,240, U.S. Pat.No. 5,216,141 and U.S. Pat.No. 4,469,863; Kiedrowshi et al., Angew. Chem. Intl. Ed. English 30: 423 (1991); Letsinger et al., J. Am. Chem. Soc. 1 10: 4470 (1988); Letsinger et al? Nucleoside & Nucleotide 13: 1597 (1994); Chapters 2 and 3, ASC Symposium Series 580, "Carbohydrate Modifications in Antisense Research", Ed. YS Sanghui and P. Dan Cook; Mesmaeker et al., Bioorganic & Medicinal Chem. Lett. 4: 395 (1994); Jeffs et al, J. Biomolecular NMR 34: 17 (1994); Tetrahedron Lett. 37: 743 (1996)) And non Non-ribose backbone described in US Pat. Nos. 5,235,033 and 5,034,506, and Chapters 6 and 7, ASC Symposium Series 580, “Carbohydrate Modifications in Antisense Research”, Ed. YS Sanghui and P. Dan Cook Including Nucleic acids containing one or more carbocyclic sugars are also included within the definition of nucleic acids (see Jenkins et al., Chem. Soc. Rev. (1995) ppl 69-176). , Rawls, C & E News Jun. 2, 1997 page 35. All of these references are hereby expressly incorporated by reference. These variations of the ribose-phosphate backbone can be made to facilitate the addition of ETMs or to increase the stability and half-life of such molecules in a physiological environment.

  As will be appreciated by those skilled in the art, all of these nucleic acid analogs can generally be used in the present invention, including use as capture and label probes. Furthermore, a mixture of naturally occurring nucleic acids and analogs can be made (eg, generally, a label probe comprises a mixture of naturally occurring and synthetic nucleotides).

  The nucleic acid can be a single helix or a double helix, as specified, or can include portions of both a double helix or a single helix sequence. The nucleic acid (especially in the case of the nucleic acid of interest) can be both DNA, gene and cDNA, RNA or hybrid, the nucleic acid can be any combination of deoxyribo-ribonucleotides, and uracil, adenine, thymine, It includes any combination of bases including cytosine, guanine, inosine, xanthine hypoxathanine, isocytosine, isoguanine and the like. One embodiment utilizes isocytosine and isoguanine in nucleic acids that are designed to be complementary to other probes rather than the sequence of interest, which reduces non-specific hybridization. This is generally described in US Pat. No. 5,681,702, the disclosure of which is hereby incorporated by reference. As used herein, the phrase “nucleoside” includes nucleotides along with nucleosides and nucleotide analogs, and modified nucleosides such as amino modified nucleosides. In addition, “nucleoside” includes non-naturally occurring analog structures. Thus, for example, the individual units of peptide nucleic acids, each containing a base, are referred to herein as nucleosides.

  As will be appreciated by those skilled in the art, a large number of analytes can be detected using this method, essentially any of the binding ligands that will be made, as described below. The analyte of interest can be detected using the method of the present invention.

  Thus, the system of the present invention is used in assaying for the analyte of interest and allows for disease diagnosis, prediction or treatment options based on the presence or absence of the analyte of interest. For example, the system of the present invention may be used for pathogen infection (bacteria (gram-positive and gram-negative bacteria, and / or the ability to distinguish them)), viruses (eg, hepatitis C virus (HCV) or respiratory viruses, It can be used for diagnosis or characterization of isotypes (including the presence or absence of viral nucleic acids), fungal infections, antibiotic drug resistance, genetic diseases (including cystic fibrosis, sickle cell anemia, etc.). Included in the definition of genetic disease for the purposes of this invention is a genetic condition that does not necessarily cause disease, but can generate other treatment options, such as one in many cytochrome p450 enzymes. Nucleotide polymorphisms (SNPs) are cases where a patient can be diagnosed as a “slow”, “normal” or “fast” digestive form. Or if the drug may be banned by a specific patient based on the patient's gene, or if the choice between two or more drugs is assisted by knowledge of the patient's gene (Such as in warfarin testing) causes different therapeutic drug digestion and leads to different dosage regimens.

[Composite cartridge]
A composite cartridge embodying aspects of the present invention is shown in FIGS. As shown in FIG. 4, the composite cartridge comprises an assembly that includes a sample preparation module 70. The sample preparation module 70 accepts, transports, mixes, mixes and performs other processes on various walls, inlet / outlet ports, fluid channels, mixing mechanisms, valves, and fluid sample material. Fluids such as reagents and buffers are treated in a manner that includes components and is described in more detail below. The sample preparation module 70 includes a substrate 72 having an upper seal 56 secured to the upper surface and a lower seal 230 secured to the lower surface. The substrate 72 includes a plurality of grooves or open channels formed in the upper and lower surfaces. Each groove may be connected to one or more inlet ports with blind holes formed in the upper surface of the substrate 72 and / or one or more outlet ports with blind holes formed in the lower surface of the substrate 72. The top seal 56 and the bottom seal 230 cover the top and bottom of the substrate 72, respectively, and have openings aligned with the inlets and outlets formed in the substrate 72 so that the conduit or sealing channel A network is formed through which fluids, such as liquids, gases, solutions, emulsions, liquid solid suspensions, etc. can flow from one part of the sample preparation module 70 to the other, inlet and outlet ports. Through which fluid can flow in and out of the sample preparation module 70, respectively. In various embodiments, the sample preparation module 70 is transparent or translucent, eg, from polycarbonate, polypropylene, acrylic, Mylar, acrylonitrile butadiene styrene (“ABS”), or other suitable polymer. Made.

  The rotary mixer 192 is operatively disposed in a mixing well 90 (described below) formed in the substrate 72. In various embodiments, the rotary mixer 192, for example, seeds a solid sample, maximizes exposure of the sample to the capture beads, mixes the sample with a chemical lysis buffer, mixes the magnetic beads with a binding buffer, and the like. (Typically, magnetic beads cannot be stored in binding buffer and therefore must only be combined at the time of use).

  A sample cap 84 is provided to surround a sample well 78 (described below) formed in the substrate 72. A plurality of deformable compartments (or blisters) 34a, 36a, 38a, 40a, 42a, and 44 are supported on top of the substrate sample preparation module 70. Each deformable compartment can contain fluid and the sample through one of the inlet ports by an openable connection that is initially sealed to prevent fluid from flowing from the blister into the channel. It can be connected to a fluid channel in the preparation module 70. When a compressive force is applied to the exterior of the blister, the increased pressure in the blister will rupture the openable connection, or otherwise open or deform, so that the fluid is removed from the blister and from the sample preparation module. Allows flow to 70 associated entrances and channels.

  The top beam 12 is disposed on the top of the cartridge on the sample preparation module 70 and includes openings of number, size and shape corresponding to the various deformable compartments supported on the sample preparation module 70. As can be seen from FIG. 1, the deformable compartments are arranged in a gap in the opening formed in the top and burr 12 so that each compartment can be compressed from above by an actuator. Provide some protection for possible compartments. In various embodiments, the top and burr 12 is provided with an inlet light port to allow monitoring of fluid movement through a particular portion of the sample preparation module 70, as described in more detail below. 14 and an exit light port 16. The top beam 12 can further include a label panel 24 on which identification information, such as human and / or machine readable instructions (eg, a barcode) can be placed.

  The top beam 12 can further include valve actuator tabs such as a sample valve actuator tab 18 and a waste valve actuator tab 20. The valve actuator tabs 18 and 20 are elastic and flexible tabs formed in the beam that will deform upon application of an external compressive force to the tabs. Each tab further includes a downwardly extending actuator post--see, eg, actuator post 26 of FIG. 1--within each sample preparation module 70, as described in more detail below, Activates an active valve located under tab 18 or 20.

  With reference to FIG. 4, the reaction module 240 is disposed below the sample processing module 70 and may be configured to receive a processed sample from the sample processing module 70 in various embodiments. In various embodiments, the reaction module 240 includes a process fluid compartment (eg, containing reagents, buffers, etc.), means for moving droplets through the module, in the indicated direction, reaction Means for culturing the mixture and means for detecting the analyte of interest (eg, nucleic acid).

  The reaction module 240 can be secured to the bottom of the sample preparation module 70 by an adhesive gasket 232 that preferably provides a leak-proof seal between the reaction module 240 and the sample preparation module 70. In various embodiments, the reaction module 240 is secured to the upper plate 241, the lower portion, and the lower portion of the upper plate 241, and defines a gap between the lower surface of the upper plate 241 and the upper surface of the fluid treatment panel 354. And a fluid treatment panel 354. This gap defines a fluid treatment and reaction space in which various steps of the assay or other process are performed.

  A lower beam 30 partially surrounds the lower portion of the cartridge assembly and cooperates with the upper beam 12 to define a relatively hard and raised outer shell of the cartridge 10. The upper and lower burrs make the cartridge 10 asymmetrical and allow the cartridge 10 to be inserted into the processing apparatus only in one direction. In the illustrated embodiment, the lower beam 30 has a rounded end 32 and the upper beam 12 has a relatively square end. Thus, the receiving slot of the processing device that receives the composite cartridge 10 and is configured to have a shape that matches the shape of the flash will ensure that the flash is always inserted to the correct side to the device. Furthermore, the lower beam 30 can include contour features such as longitudinal side grooves 22 that extend only partially along the length of the lower beam 30. Such grooves cooperate with corresponding features in the receiving slot of the processing device to ensure that the cartridge is inserted into the device in the proper orientation.

Deformable fluid compartment (blister)
In general, a blister is made of a deformable material that preferably bursts when an appropriate pressure is applied; that is, the material used to form the blister is its original material when the pressure is removed. It does not return to the shape. This is because it can cause a back flow of the added reagent. In addition, blisters can be used once (one application of pressure is made during the assay) or several times (eg, multiple aliquots of reagents can be used in a single location or multiple locations during the run of the assay). Can be used). Each blister can contain a unique process material (eg, buffer, reagent, insoluble liquid, etc.), or two or more blisters can contain the same process material. This redundancy can be used to give the same process material to multiple locations in the disposable rest.

  Although the size, number, arrangement, and content of the compartments are largely determined by the assay or other process that is intended to be performed on the composite cartridge 10, the illustrated embodiment is directed to six deformable fluid compartments. Or blisters 34a, 36a, 38a, 40a, 42a, and 44. The deformable blister can have an associated lance blister. In the illustrated embodiment, each of the deformable fluid blisters 34a, 36a, 38a, 40a, and 42a has an associated deformable lance cartridge or lance blister 34b, 36b, 38b, 40b, and 42b.

  The operation of the deformable compartment embodiment will be described with reference to FIG. 5, which shows a cross section of the deformable compartment 34a. In various embodiments, the deformable compartment of the composite cartridge 10 incorporates the features described in the co-owned US patent application Ser. No. 14 / 206,867, entitled “Devices and Methods for Manipulating Deformable Fluid Vessels”. This content is incorporated herein by reference.

  When compressing the deformable compartment and moving its fluid contents, sufficient compressive force is applied to the blister to rupture the breachable seal holding the fluid in the compartment, or other If so, it must be opened. The amount of force required to break the seal and move the fluid content of the compartment typically increases as the volume of the compartment increases. To limit the amount of compressive force that must be applied to the deformable compartment or blister to rupture or otherwise open the breachable seal holding the fluid in the compartment. A blister 34b is provided in connection with the deformable compartment 34a. The deformable compartment 34a and the lance blister 34b can be connected by a channel that can be blocked initially by a rupturable seal. The lance blister 34b is a fluid port 136 formed in the sample preparation module 70 by, for example, a deformable foil split or septum that holds the bead 46 and the fluid contents in the lance blister 34b and the deformable compartment 34a. It includes an opening device such as a bead 46 (such as a steel ball bearing) supported above and enclosed within a lance blister 34b. Thus, in order to open the deformable compartment 34a, first, a bead is applied via a foil split that blocks the fluid port 136 by applying a compressive force to the lance blister 34b from the outside to compress the lance blister 34b. Apply force to 46. After the fluid port 136 is opened, the fluid contents of the deformable compartment 34a can be sent into the fluid port 136 relatively easily by application of an external compressive force to the deformable compartment 34a. The amount of pressure required to compress the lance blister 34b and apply force to the bead 46 through the foil split will generate enough pressure to compress the main compartment 34a and open the breachable seal. Much smaller than required. The fluid that flows into the fluid port 136 is then formed in the lower surface of the substrate 72 and defined by a groove covered by the lower seal 230, through the horizontal channel 137 to and from the vertical channel transition 139. Will flow to one or more other points in the sample preparation module 70.

[Sample preparation module]
Various details of the sample preparation module 70 are shown in FIGS.

  Sample well 78 is configured to receive a fluid sample material to be assayed or otherwise processed in composite cartridge 10. As shown in FIGS. 6 and 7, the sample well 78 can be defined by a vertical peripheral wall 79 (circular in the illustrated embodiment) and a lower wall or floor 81. The sample well 78 further includes an entrance snorkel 80 that extends along the peripheral wall 79 of the sample well 78 and ends at a position below the top of the peripheral wall. The exit port 82 is provided on the floor 81 of the well 78, and the floor 81 is preferably conical so as to taper downward toward the exit port 82.

  A sample cap 84 can be provided to seal the sample well 78 after the sample material has been placed in the sample well 78. In one embodiment, the sample cap 84 comprises a circular cover having an outer peripheral wall that fits over the vertical peripheral wall 79 of the sample well 78. The sample cap 84 is defined by a radial resilient locking tab that extends through an opening in the substrate 72 such that the cap 84 is pivotally coupled to the sample well 78 about an axis defined by the pivot post 86. A pivot post 86 can be included. After the sample material is placed in the sample well 78, the sample cap 84 can be pivoted onto the top of the sample well 78 and pushed down onto the sample well 78. A clip or other detent 88, extending upwards, catches the sample cap 84 when it is pushed down into the clip 88, locks it tightly, and confirms that the cap 84 is closed tightly. Can be provided as can be. In some embodiments, the sample cap 84 can have a lower surface that tapers downward when the sample cap 84 is positioned over the sample well 78 (not shown). The conical structure helps to reduce the amount of fluid aggregation retained on the inner surface of the sample cap 84 during processing of the sample in the sample well 78.

  Sample preparation module 70 also includes a mixing well 90 formed in substrate 72. As shown in FIGS. 8A and 9A, the mixing well 90 can be defined by a vertical peripheral wall 91 (circular in the illustrated embodiment) and a lower wall or floor 93. In various embodiments, the fluid inlet snorkel 92 extends to the peripheral wall 91 of the mixing well 90 and ends below the top of the wall 91. In various embodiments, the pressure snorkel 94 extends to the other part of the peripheral wall 91 of the mixing well 90 and ends at a position below the top of the wall 91. The exit port 96 allows fluid to exit the mixing well 90 and is located near the center of the tapered portion of the wall 90 below the floor 93 and is formed in the lower center of the floor 93. A plurality of openings surrounding the sheet 98 can be provided.

  The rotary mixer 192 includes an upper circular disc 194 disposed in the mixing well 90 and supported on the upper end of the peripheral wall 91 of the wall 90. The peripheral gear teeth 198 are formed around the periphery of the disk 194, and a portion of the teeth 198 protrudes from the upper and lower ends of the composite cartridge 10 and the outer ends of the flash beams 12, 30 and provides power to the rotation of the rotary mixer 192. Therefore, it can be engaged with the external drive mechanism of the processing apparatus. An O-ring 196 is disposed in the peripheral O-ring groove around the upper disk 194 below the peripheral gear teeth 198. The O-ring 196 closes the space between the rotary mixer 192 and the peripheral wall 91 of the wall 90. The spindle 200 extends downward from the upper disk 194 and is secured within the central spindle sheet 98 of the mixing well 90. The plurality of impeller blades 202 extend radially from the spindle 200.

  Another embodiment of a mixing well 90 'is shown in FIGS. 8B and 8C. As shown, the mixing well 90 'can be defined by a vertical peripheral wall 91' (circular in the illustrated embodiment) and a lower wall or floor 93 '. The fluid inlet snorkel 92 'extends to the outer surface of the peripheral wall 91' of the mixing well 90 'and includes an opening 92a below the top of the wall 91'. The pressure snorkel 94 'extends to the outer surface of the peripheral wall 91' of the mixing well 90 'and includes an opening 94a below the top of the wall 91'. The exit port 96 'allows fluid to exit the mixing well 90' and is located near the center of the tapered portion of the wall 90 'on the underside of the floor 93', and the lower center of the floor 93 ' A plurality of openings surrounding the spindle sheet 98 ′ formed on the substrate can be provided. The exit port 96 ′ and spindle sheet 98 ′ can be substantially identical to the exit port 96 and spindle sheet 98, respectively, of the mixing well 90.

  In another mixing well 90 ′ of FIGS. 8B and 8C, the rotary mixer disposed within the mixing well 90 ′ has impeller blades extending radially from the mixer spindle to substantially the inner surface of the peripheral wall 91 ′. Can be configured. This provides a clearance for the snorkels 92, 94 formed on the inner surface of the peripheral wall 91 so that the radial impeller blades 202 cannot extend substantially to the inner surface of the peripheral wall 91. This is contrary to the structure of the rotary mixer 192 configured for operation. Having a mixer with impeller blades extending to the inner surface of the peripheral wall 91 'provides more complete and / or effective mixing of the contents of the mixing well 90' in some situations.

  Referring to FIG. 9B, the rotary mixer 192 ′ disposed within the mixing well 90 ′ is substantially identical to the circular disk 194, peripheral gear teeth 198, and O-ring 196 of the rotary mixer 192 shown in FIG. 9A. An upper circular disc 194 ′, peripheral gear teeth 198 ′, and an O-ring 196 ′ may be included. The spindle 200 'extends downward from the upper disk 194'. Two or more impeller blades 202 'extend radially from the spindle 200'. The impeller blade 202 'extends substantially to the inner surface of the peripheral wall 91'. In various embodiments, the impeller blade 202 'can be distorted relative to the spindle 200' and further includes an opening 203 formed therein to improve the mixing efficiency of the rotary mixer 192 '. Can do.

  Referring again to FIG. 15, which shows a top plan view of the sample preparation module 70, the sample preparation module 70 can include alignment holes 74 and 76, or other alignment features can be located within, for example, pins or each alignment hole. Other structures within the apparatus that extend into the sample can be provided in the sample preparation module 70 or some other portion of the composite cartridge 10 to facilitate alignment of the composite cartridge 10 with the processing apparatus.

  The sample preparation module 70 includes a first inlet port 136 formed in the upper surface of the module through which process fluid from the deformable compartment 34a can be introduced into the sample preparation module 70. In one embodiment, the deformable compartment 34a is commercially available, such as those containing a lysis buffer such as water for low osmotic dissolution or a chiatropic salt such as a guanidinium salt. Lysis buffer, and / or high / low pH, and / or sodium dodisyl sulfate (SDS), TWEEN® 20 (polysorbate 20), TRITON ™ X-100 (polyoxyethylene octyl phenyl ether) Including a surfactant. In some cases, the lysis buffer optionally includes reagents to interfere with undesired enzyme activities such as DNase and RNase activity (which depend on the analyte and assay of interest, This can then be added as needed, although it can be either a dry or liquid individual reagent), which is then removed during the bead capture / elution process.

  After the sample material cells are lysed, it is often desirable to perform at least partial purification to remove other cells and sample debris from the sample to facilitate downstream handling and processing. . The research sample in the buffer does not necessarily need to be purified, but even so, purification is typically performed. Existing technology relies on the use of target capture beads (eg, magnetic capture beads) to capture and immobilize desired analyte (s) of interest from cells and sample debris. In various implementations, the capture beads and binding buffer are mixed with the sample in the lysis buffer after cells or viruses have been disrupted by mechanical and / or chemical means. The capture bead can have magnetism to facilitate substantial immobilization of the bead and the analyte of interest bound thereto by selective application of magnetic force, as will be appreciated by those skilled in the art. As such, other implementations can use non-magnetic beads such as polystyrene or silica beads (eg, the beads can be captured by size or in an area in the affinity column).

  Thus, in various embodiments, the sample preparation module 70 includes a second inlet port 138 through which process fluid can be introduced into the sample preparation module 70 from the deformable compartment 36a. In one embodiment, the deformable compartment 36a includes a binding buffer to promote binding of a target capture bead, such as a magnetic bead, with one or more analytes of interest.

  In various embodiments, the sample preparation module 70 includes a third inlet port 140 through which process material from the deformable compartment 44 can be introduced into the sample preparation module 70. In one embodiment, the deformable compartment 44, in combination with a binding buffer from the deformable compartment 36a, binds to one or more analytes of interest within the sample material, thereby providing sample material. The target capture beads that can be equipped with magnetic particles that isolate the analyte (s) of interest from the remainder and that allow magnetic separation.

  The capture beads can be coated with a material that facilitates capture of the analyte (s) of interest. For example, for nucleic acid capture, the beads can be coated with a negatively charged coating to facilitate absorption of the positively charged nucleic acid relative to the surface, then washed with a buffer, and then Treated with elution buffer to remove the purified nucleic acid from the beads for further processing. As will be appreciated by those skilled in the art, a number of suitable commercial products including, for example, MagaZorb® beads from Promega, MagMax from Life Tech, or beads from Qiagen, MoBio, BioRad, etc. There are bead systems that are commercially available.

  Thus, the capture beads of interest that can be included in the deformable compartment 44 are in fluid access to a binding buffer, such as a binding buffer that can be included in the deformable compartment 36a used with the capture beads. Facilitates purification of analytes of interest.

  In another embodiment, the capture beads of interest are within the mixing well 90 during assembly of the composite cartridge 10, for example, until reconstituted by fluid added to the mixing well 90 during use of the composite cartridge 10. Can be provided directly in the sample preparation module 70 in the form of a lyophilized pellet that is placed in a pellet and stored in a mixing well.

  In another implementation, the capture beads can be functionalized with a capture nucleic acid probe to extract the nucleic acid either specifically or non-specifically. For example, beads can generally be functionalized with random 6-mers, or with capture probes specific for the desired nucleic acid of interest, to extract nucleic acids. In some cases, for example, when mRNA is of interest, beads coated with poly-T capture probes can be used.

  In various embodiments, the sample preparation module 70 further includes a fourth inlet port 142 through which process material from the deformable compartment 38a can be introduced into the sample preparation module 70. In one embodiment, the deformable compartment 38a includes an insoluble fluid (eg, an oil such as mineral oil, silicone oil, as discussed in detail below).

  In various embodiments, the sample preparation module 70 further includes a fifth inlet port 144 through which process material from the deformable compartment 40 a can be introduced into the substrate 72. In one embodiment, the deformable compartment 40a includes an elution buffer.

  In various embodiments, the sample preparation module 70 further includes a sixth inlet port 146 through which process material from the deformable compartment 42a can be introduced into the sample preparation module 70. In one embodiment, the deformable compartment 42a includes a wash buffer.

  In various embodiments, the sample preparation module 70 includes a first outlet port 182, a second outlet formed on the lower surface of the sample preparation module 70 that allows fluid to exit the module 70 and flow into the reaction module 240. Outlet port 188 and a third outlet port 190.

  Designating an entry port or exit port as a first, second, third, fourth, fifth, or sixth port merely provides a convenient means of distinguishing each port from each other. It should be noted that there is no limitation and is not meant to be limiting, for example, by specifying a particular order or sequence in which ports can be used.

  The first fluid channel 150 extends from the first inlet port 136 to the sample well 78. In the figure, the fluid channel is represented by parallel lines extending between two points across the sample preparation module 70. Each channel can include one or more channel transition points, represented by circles within the channel, one of which is indicated by reference numeral 151. The channel transition point is from a channel portion formed at the bottom of the substrate 72 to a channel portion formed at the top of the substrate 72, or from a channel portion formed at the top of the substrate 72 to a channel formed at the bottom of the substrate 72. Represents a vertically extending portion of a channel that extends into the portion and allows the channel to pass over or under other channels in the substrate 72.

  Second fluid channel 152 extends from sample well 78 to lysis chamber inlet 122. The third fluid channel 156 extends from the lysis chamber outlet 124 to a fifth fluid channel 162 that extends from the third inlet port 140 to the mixing well inlet snorkel 92. The fourth fluid channel 160 extends from the second inlet port 138 to the third inlet port 140. The sixth fluid channel 164 extends from the fifth inlet port 142 to the first outlet port 182. The seventh fluid channel 166 extends from the fifth inlet port 144 to the second outlet port 188. An eighth fluid channel 168 extends from the mixing well exit port 96 to a passive valve assembly 220 (described below). The ninth fluid channel 170 extends from the passive valve cavity of the passive valve assembly 220 to the capture compartment 100. Tenth fluid channel 172 extends from active valve assembly 204 to active valve assembly 219. An eleventh fluid channel 174 extends from the active valve assembly 219 to the waste chamber 102. A twelfth fluid channel 176 extends from the sixth inlet port 146 to the capture compartment 100. A thirteenth fluid channel 178 extends from the capture compartment 100 to the active valve assembly 204. Fourteenth fluid channel 180 extends from active valve assembly 204 to third outlet 190.

  Designating various fluid channels as first, second, third, fourth, fifth, etc. fluid channels only provides a convenient means of distinguishing each port from each other. It is noted that there is no implied limitation, eg, by specifying a particular order or sequence in which the fluid channels can be used, or a particular direction in which fluid flows through the channels. Should.

  In various embodiments, the sample preparation module 70 further includes a passive valve assembly 220 adjacent to the mixing well 90. In one embodiment, the passive valve assembly 220 is closed so that if the pressure in the mixing well 90 is below a threshold pressure, thereby retaining fluid in the mixing well 90, the passive valve assembly 220 is closed. Composed. On the other hand, if the pressure can be increased within the mixing well 90 to a sufficient pressure level that is greater than the threshold pressure, the passive valve assembly 220 is opened, thereby causing the exit port 96 and the mix well exit. Fluid in the mixing well is allowed to escape via an eighth fluid channel 168 connecting port 96 to passive valve assembly 220.

  Details of the passive valve assembly 220 are shown in FIGS. The valve assembly 220 includes a valve cavity 222 formed in the substrate 72 and an inlet 224 formed in the substrate 72 and extending upward into the valve cavity 222. The valve 229 can include a Belleville valve, but is disposed in the valve cavity 222 via the inlet 224. The cage 226 is disposed on the valve 229. The outlet 228 extends radially from the valve cavity 222.

  Under conditions where no pressure is applied, the valve 229 and the retainer 226 are seated below the valve cavity 222 with the valve 229 covering the inlet 224. The cage 226 can be biased to a down position, for example, by a suitable spring or the like. Accordingly, fluid flowing from the inlet 224 cannot pass into and through the valve cavity 222, and therefore fluid cannot escape the mixing well 90. On the other hand, when the fluid in the inlet 224 is under sufficient pressure (eg, about 3-5 psi) to exceed any force (eg, spring bias) holding the retainer 226 in the down position, the valve 229 and retainer 226 are lifted from the bottom of the valve cavity 222, thereby opening the inlet 224 and allowing fluid to flow into the valve cavity 222 and out of the outlet 228.

  The sample preparation module 70 can further include a pump port 104 to which an external source of pressure can be coupled to the sample preparation module 70. The pump port 104 is connected to the sample well 78 via the pressure conduit 106, and the pressure applied to the pump port 104 pressurizes the sample well 78 and allows the contents of the sample well 78 to exit the well. Let

  The sample preparation module 70 can further include a passive valve port 108 connected to the pressure snorkel 94 of the mixing well 90 via a valve conduit 110. If the passive valve port 108 is open, the pressure will increase in the mixing well 90 and the passive valve assembly 220 will remain closed. When the passive valve port 108 is closed, the pressure will increase in the mixing well 90 and the passive valve assembly 220 will open so that the contents of the mixing well 90 can flow out of the well.

  Some organisms such as viruses and many bacteria can be chemically lysed by adding a lysis buffer with or without elevated temperature or proteolytic enzymes. Some organisms are difficult to dissolve by chemical and / or enzymatic methods and require mechanical disruption or shearing of the cell membrane. Therefore, an optional component of the composite cartridge 10 is an impeller component that imposes a solid component so that individual cells can be accessed by the lysis buffer and to release more analyte of interest. Fired to crush or destroy. The impeller imparts turbulent action to the fluid containing the dissolved beads. The main lysis action is that the beads are lysed by collision with the target organism, breaking them open and exposing the nucleic acid of interest. The presence of lysis buffer suppresses DNase or RNase, which can destroy RNA or DNA objects if cells are destroyed. In various embodiments, the impeller appears to be a very fast rotating outer ring.

  Thus, in various embodiments, the sample preparation module 70 further includes a lysis chamber 120 having a driven agitator, such as a motor driven bead mixer mechanism, disposed therein. The drive agitator is disposed and arranged at least partially within the lysis chamber 120 and is configured to agitate the fluid flowing through the processing chamber. The fluid flowing through the lysis chamber can include a mixture of sample material, lysis buffer, and lysis beads. The lysis beads can include silica (ceramic) beads (eg, 100 μm diameter) that are fed into the lysis chamber 120 during assembly of the composite cartridge 10. The bead mixer includes a motor 128 having an impeller 130 mounted on the output shaft of the motor (see FIG. 2). The fluid flows through the inlet 122 into the lysis chamber 120 and out of the lysis chamber 120 through the outlet 124. The mesh filter can be provided in front of the inlet 122 and / or the outlet 124. The mesh filter (s) has a hole size configured to hold the lysis beads in the lysis chamber 120 as the sample fluid flows into and out of the lysis chamber 120. In operation, the motor 128 causes the impeller 130 to rotate at a high rotational speed (eg, from about 5,000 to about 100,000 rpm, preferably from about 10,000 to about 50,000 rpm, more preferably from about 20,000 to about The fluid in the lysis chamber 120, which can be rotated at 30,000 rpm) and can contain sample material and lysis beads, is vigorously agitated by the rotating impeller, thereby dissolving the molecular structure of the sample material. Help beads. Thus, the sample mixture flowing out of the lysis chamber 120 is more completely lysed than without the bead mixer.

  A suitable motor 128 for the bead mixer includes Feying, Model FY0610-Q-04170Y from Jinlong Machinery. The motor can be powered by a temporary connection of the composite cartridge 10 to an external power source of the device where the cartridge 10 is processed. Control of the motor 128 can be implemented by logic elements provided outside and / or inside the cartridge 10. In one embodiment, a mixer printed circuit board (“PCB”) is provided in the lower beam 30 that controls the operation of the bead mixer motor 128. The mixer motor 128 is ideally operated only when fluid flows through the lysis chamber 120. Fluid flowing into the lysis chamber 120 is detected by a light sensor through an inlet light port 14 formed in the top beam 12 (see FIG. 2) that is aligned with the inlet light sensing chamber 154 (see, eg, FIG. 15). The bead mixer motor 128 can be activated, for example, upon detection of the front end of the fluid stream flowing through the inlet light sensing chamber 154 toward the lysis chamber 120. Similarly, fluid flowing out of the lysis chamber 120 can be detected by a light sensor via an exit light port 16 (see FIG. 2) aligned with the exit light sensing chamber 158 (see FIG. 15), and the bead mixer The motor 128 can be deactivated, for example, upon detection of the trailing edge of the fluid stream flowing through the exit light sensing chamber 158.

  Sample preparation module 70 further includes two active valve assemblies 204, 219. The valve assembly 204, known as the sample valve assembly, is located at the junction of the tenth fluid channel 172, the thirteenth fluid channel 178, and the fourteenth fluid channel 180 and is connected to the thirteenth fluid channel 178 through the fourteenth fluid channel. Controls flow to the fluid channel 180. The valve assembly 219 is known as a waste valve assembly and is located at the junction of the tenth fluid channel 172 and the eleventh fluid channel 174 and from the tenth fluid channel 172 to the eleventh fluid channel 174 and the disposal chamber 102. To control the flow.

  Details of the active valve assembly, such as the bubble assembly 204, are shown in FIGS. The valve assembly 204 includes a valve cavity 210 formed in the substrate 72. An inlet conduit 208 leads to the valve cavity 210 and an outlet channel 212 extends from the cavity 210. Access opening 206 is formed in top seal 56 disposed on top of substrate 72. The flexible valve membrane 216 is secured to the underside of the upper seal 56 below the access opening 206 by an adhesive 214 surrounding the access opening 206. As shown in FIG. 13, in an unactuated or unactuated position, fluid flows into the valve cavity 210 through the inlet 208 and out of the valve cavity 210 through the outlet 212. Can do. Thus, the fluid flowing through the valve assembly 204 is smooth. As shown in FIG. 14, fluid flowing through the valve assembly 204 is blocked when the external valve actuator 218 pushes down through the access opening 204 to deform the valve membrane 216 on the outlet 212. The valve actuator 218 can include an actuator post 26 of an actuator tab 20 formed on the top beam 12 (see FIG. 1). In particular, the valve actuator tab 18 is aligned with the active valve assembly 204 and the valve actuator tab 20 is aligned with the active valve assembly 219.

  In various embodiments, the sample preparation module 70 further includes a waste chamber 102 (or one or more waste chambers) configured to receive and store excess or spent fluid.

Reaction Module-Top Plate Details of the reaction module 240 and top plate 241 are shown in particular in FIGS. Referring to FIGS. 24 and 26, which show a perspective view and a top plan view, respectively, of the upper plate 241, the upper plate 241 has an upper peripheral wall 256 protruding on the upper surface 242 of the upper plate 241, and an upper plate 241. And an upper surface 240 that at least partially surrounds the periphery at a position that is offset inwardly from the outer end of the. The upper peripheral wall 256 has a continuous open channel or groove 258 formed along the upper end that provides a mounting location for the adhesive gasket 232 that secures the reaction module 240 to the sample preparation module 70. See FIG. The upper peripheral wall 256 forms a recessed area 260 that is surrounded by the upper peripheral wall 256 on the upper surface 242. See also FIG. 28 and FIG.

  The upper plate 241 can adopt a plurality of configurations and can be made of various materials. Suitable materials include, but are not limited to, fiberglass, TEFLON®, ceramic, glass, silicon, mica, plastic (acrylic, polystyrene and copolymers of styrene and other materials, polypropylene, polyethylene, polybutylene, Polycarbonate, polyurethane, and derivatives thereof). A particularly suitable top plate material is polycarbonate.

  The alignment fork 246 extends from one end of the upper plate 241 and the alignment loop 244 extends from the other end of the upper plate 241. The alignment fork 246 and the alignment loop 244 are configured to receive an alignment pin of the apparatus for processing the composite cartridge 10 to ensure proper alignment of the cartridge 10, as will be described in more detail below.

  The upper plate 241 further includes a sample compartment 266 having an inlet port 268 that is in fluid communication with the third outlet port 190 of the sample preparation module 70.

  The top plate 241 further includes a rehydration (elution) buffer compartment 276 having a second outlet port 188 of the sample preparation module 70 and an inlet port 278 through which fluid is passed. The detection buffer compartment 280 is sent to the rehydration buffer compartment 276 and forwarded to the detection buffer compartment 280, the first dry detection buffer reconstituted with an amount of reconstitution buffer. (Applies to a portion of the top plate 241 that forms the detection buffer compartment 280 or a portion of the fluid treatment panel 354 that covers the detection buffer compartment 280). In one embodiment, the detection buffer compartment 280 has a volume of 120-160 μl. In various embodiments, the top plate 241 includes a connecting passage 274 between the detection buffer compartment 280 and the rehydration buffer compartment 276. The detection buffer compartment 280 can further include a port 282 for injecting buffer into the compartment 280 during the manufacturing process and / or to vent the compartment 280.

  25 and 27 show a lower perspective view and a lower plan view of the upper plate 241, respectively. Referring to FIGS. 25 and 27 in addition to FIGS. 24 and 26, the top plate 241 is later reconstituted in one embodiment by an amount of rehydration buffer from the rehydration buffer compartment 276. (Rehydrated) in a dry form of PCR buffer / enzyme (part of top plate 241 forming buffer compartment 296 or fluid treatment panel 254 covering buffer compartment 296 (FIGS. 4 and 58)), which further includes a buffer compartment 296. In one embodiment, the buffer compartment 296 has a volume of about 20 μl. Port 298 is provided for injecting PCR buffer / enzyme into the compartment during the manufacturing process and / or to vent the buffer compartment 296.

  The upper plate 241 is rehydrated later by an amount of rehydration buffer from the rehydration buffer compartment 276, the dried form of exonuclease reagent (the upper plate 241 forming the second buffer compartment 300). Or a second buffer compartment 300 that may be applied to a portion of the fluid treatment panel 354 that covers the second buffer compartment 300. In one embodiment, the second buffer compartment 300 has a volume of about 20 μl. Port 302 may be provided for injecting buffer into the second buffer compartment 300 during the manufacturing process and / or to vent the compartment 300. A dam 306 may be provided between the rehydration buffer compartment 276 and the second buffer compartment 300 to allow fluid to flow from the rehydration buffer compartment 276 to the compartment 300. .

  The upper plate 241 further includes a lower peripheral wall 290 that surrounds the periphery of the lower portion of the upper plate 241. The lower peripheral wall 290 defines a recess surrounded by a peripheral wall 290 configured to receive a panel, such as a fluid treatment panel 354, to surround the lower half of the upper plate 241. The rising panel support 290 surrounds the outer periphery of the lower surface of the upper plate 241 just inside the peripheral wall 290. The area 294 inside the panel support 292 is arranged with a slight gap with respect to the panel support 292, the panel inserted into the peripheral wall 290 is supported on the panel support surface 292, and the recess 294 is formed in the panel support 292. A gap 295 (see FIGS. 28 and 29) is defined between the upper plate 241 and the upper plate 241.

  The top plate 241 can further include fluid inlet ports 250, 252, at least one of which is in fluid communication with the first outlet port 182 of the sample preparation module 70. Inlet ports 250, 252, for example, provide a fluid path between gap 295 between the lower surface of reaction upper plate 241 in region 294 and a fluid treatment panel 354 surrounding the lower surface of upper plate 241.

  The top plate 241 further includes detection compartments 350a, 350b, 350c, and 350d, each having an inlet port or vent port 352. Although the illustrated embodiment includes four detection compartments 350a-d, other configurations of the top plate 241 with fewer or more detection compartments 350 are readily conceivable.

  The area 304 on the lower surface includes a processing area that is slightly spaced from the area 294, so that it is larger than the area 294 between the upper plate 241 and the lower panel in the area 304. Create a gap.

  The reaction module 240 can further include one or more bubble traps 340 formed in the upper plate 241. Each bubble trap 340 includes a bubble capture hood 342 formed in an upper plate 241 that slopes upward toward the vent opening 344. In one embodiment, rising air bubbles generated by fluid movement under the bubble trap are captured by the capture hood 344 and released through the vent opening 344. The capture hood can be shaped to fit a fluid movement path below or adjacent to the bubble trap. In the illustrated embodiment, as described in more detail below, five bubble traps 340 with elongated capture hoods 342 are disposed on four fluid movement paths, each of which is adjacent to two adjacent bubble traps 340. And between.

  Details of the fluid inlet 252 are shown in FIG. As described, the fluid inlet 252 can be aligned with the first fluid outlet 182 of the sample preparation module 70. The fluid inlet 252 is tapered inward and has a frustoconical shape, and the size of the outlet 182 above the inlet 252 is narrower than the upper end of the inlet 252. This helps to ensure that the fluid discharged from the outlet 182 is captured by the inlet 252.

  Details of sample compartment 266, rehydration buffer compartment 276, and detection buffer compartment 280 are shown in FIG. Sample compartment 266 is configured to receive an amount (eg, 200 μl) of magnetic beads with bound target analyte (eg, DNA, nucleic acid) from sample preparation module 270 via inlet port 268. Is done. The inlet port 268 of the sample compartment 266 preferably has a conical shape and is aligned with the third outlet 190 of the sample preparation module 70. In various embodiments, the third outlet 190 passes through a tapered nipple 322 to minimize drop dripping from the end of the outlet 190. The inlet port 268 is also preferably tapered at its widest end at the top, thereby ensuring that fluid discharged through the outlet 190 is captured at the inlet port 268. . The outlet 190 and the inlet port 268 are configured such that there is a small gap between them. This gap comprises a portion of the interstitial space 308 between the upper part of the upper plate 241 of the reaction module 240 and the lower part of the sample preparation module 70. This gap provides a trap for collecting any air bubbles contained in the fluid within the reaction module 240, in particular, air bubbles that may be generated when exhausting fluid from the outlet 190 to the inlet port 268.

  Rehydration buffer compartment 276 receives an amount (eg, 200 μl) of buffer solution suitable for rehydration of dry reagents and elution of nucleic acids from beads via inlet port 278 from sample preparation module 270. Configured as follows. The inlet port 278 of the rehydration buffer compartment 276 is aligned with the second outlet 188 of the sample preparation module 70. Again, the outlet 188 preferably flows through the tapered nipple 320 and its end is spaced from the similarly tapered inlet port 278. Again, the space between the end of the nipple 320 and the inlet port 278 allows gas bubbles in the fluid flowing between the outlet 188 and the inlet port 278 to escape to the intervening space 308.

[Reaction Module-Fluid Processing Panel]
Referring to FIGS. 58 and 59, in various embodiments, the reaction module 240 of the composite cartridge 10 includes a fluid treatment panel 354 secured to the lower portion of the upper plate 241. The fluid treatment panel 354 is surrounded by a peripheral wall 290 and supported and fixed on the panel support 292 by, for example, oil and a temperature resistant adhesive. The fluid treatment panel 354 facilitates multiple functions of the composite cartridge 10, such as fluid movement and analyte detection. Such fluid movement may involve transporting one or more droplets of fluid along a fluid transport path (eg, back and forth linearly or continuous (eg, circular, elliptical, square)). Mixing fluids by moving one or more droplets to vibrate (on the path), combining fluid droplets that can contain different materials, and dropping droplets into two or more small droplets Splitting, etc. can be included.

  The fluid treatment panel 354 includes a substrate 356. Suitable substrates include metal surfaces such as gold, electrodes as defined below, glass and modified or functionalized glass, fiberglass, ceramic, mica, plastics (acrylic, polystyrene and copolymers of styrene and other materials, polypropylene, Including polyethylene, polybutylene, polyimide, polycarbonate, polyurethane, TEFLON (registered trademark), and derivatives thereof), GETEK (registered trademark) (mixed of polypropylene oxide and fiberglass), polysaccharide, nylon or nitrocellulose, Resins, silica or silica-based materials including silicon and modified silicon, carbon, metals, inorganic glasses and various other polymers, and particularly suitable printed circuit board (PCB) materials.

  In various embodiments, the fluid treatment panel 354 can be divided into a plurality of different functional areas or treatment areas, which are spatially overlapping or spatially different, or It can be partly spatially separate and partly spatially different.

  In various embodiments, the fluid reaction process in the reaction module 240 is based on microfluidic fluid manipulation using so-called electrowetting techniques to form microdroplets that can be manipulated both spatially and biochemically. Based at least in part.

  In general, electrowetting is to modify the wetting characteristics of a hydrophobic surface (such as PCB) by an applied electric field. In electrowetting systems, the change in substrate-electrolyte contact angle due to the applied potential difference results in the ability to move the electrolyte on the surface. In essence, as described in US Pat. No. 6,565,727, this disclosure is expressly incorporated herein by reference, but with polar liquids (eg, those containing the analyte of interest). By applying a potential to the electrodes (or collection of electrodes) adjacent to the droplets, the surfaces on these electrodes become more hydrophilic and the droplets increase the overlap area with the charged electrodes. Because of the surface tension gradient. This allows the droplet to spread over the surface and then remove the potential or activate a different electrode to return the substrate to a hydrophobic state, resulting in a new hydrophilicity on the substrate. The liquid will move to the area. Thus, for processing, handling, and detection, droplets can be moved physically and individually to different processing areas on the flat surface of the substrate. The droplets can be moved at various speeds and split (eg, a single droplet can be split into two or more droplets), pulsed, and / or mixed (Two or more droplets can be combined at the same location and then split or moved as one). Furthermore, electrowetting can cause mixing within a single droplet. As described in more detail below, the droplets can also be used to rehydrate dry reagents stored at different locations on the PCB substrate. One typical characteristic of electrowetting is the precise manipulation of very small fluid volumes. For example, an isolated nucleic acid of interest can be obtained at a very high concentration of less than 10 μl prior to PCR amplification, compared to an elution volume of 100 μl and a much lower concentration of the analyte of interest characteristic of other systems. Can be eluted. In addition, electrowetting eliminates the need to make any changes to the physical interface (eg, new valves, fluid pathways, etc.), allowing fluid pathways to be changed during development and in the product. And

  Exemplary microfluidic systems using electrowetting techniques are described in U.S. Patent Publication No. 2013/0252262, U.S. Patent Publication No. 2013/0233712, U.S. Publication No. 2013/0233425, U.S. Patent Publication No. 2013/0230875, US Patent Publication No. 2013/0225452, US Patent Publication No. 2013/0225450, US Patent Publication No. 2013/0217113, US Patent Publication No. 2013/0217103, US Patent Publication No. 2013/0203606, US Patent Publication No. 2013 / 0178968, US Patent Publication No. 2013/0178374, US Patent Publication No. 2013/0164742, US Patent Publication No. 2013/0146461, US Patent Publication No. 2013/0130936, US Patent Publication No. 2013/01 18901, US Patent Publication No. 2013/0059366, US Patent Publication No. 2013/0018611, US Patent Publication No. 2013/0017544, US Patent Publication No. 2012/0261264, US Patent Publication No. 2012/0165238, US Patent Publication No. 2012 / 0132528, US Patent Publication No. 2012/0044299, US Patent Publication No. 2012/0018306, US Patent Publication No. 2011/0311980, US Patent Publication No. 2011/0303542, US Patent Publication No. 2011/0209998, US Patent Publication No. 2011/0203930, US Patent Publication No. 2011/0186433, US Patent Publication No. 2011/0180571, US Patent Publication No. 2011 / 01 14490, US Patent Publication No. 2011/0104816, US Patent Publication No. 2011/0104747, US Patent Publication No. 2011/0104725, US Patent Publication No. 2011/0097763, US Patent Publication No. 2011/0091989, US Patent Publication No. 2011/0086377, US Patent Publication No. 2011/0076692, US Patent Publication No. 2010/0323405, US Patent Publication No. 2010/0307917, US Patent Publication No. 2010/0291578, US Patent Publication No. 2010 / 0282608, US Patent Publication No. 2010/0279374, US Patent Publication No. 2010/0270156, US Patent Publication No. 2010/0236929, US Patent Publication No. 2010/0236928, US Patent Publication No. 2010/0206094, US US Patent Publication No. 2010/0194408, US Patent Publication No. 2010/0190263, US Patent Publication No. 2010/0130369, US Patent Publication No. 2010/0120130, US Patent Publication No. 2010/0116640, United States Patent Publication No. 2010/0087012, United States Patent Publication No. 2010/0068764, United States Patent Publication No. 2010/0048410, United States Patent Publication No. 2010/0032293, United States Patent Publication No. 2010/0025250 No., US Patent Publication No. 2009/0304944, US Patent Publication No. 2009/0263834, US Patent Publication No. 2009/0155902, US Patent Publication No. 2008/0274513, US Patent Publication No. 2008/0230386, US Patent Publication No. 2007/0275415, U.S. Patent Publication No. 2007/0242105, U.S. Patent Publication No. 2007/0241068, U.S. Patent No. 8541176, U.S. Patent No. 8492168, U.S. Patent No. 8481125, U.S. Pat. 8460528, U.S. Patent No. 8454905, U.S. Patent No. 8440392, U.S. Patent No. 8426213, U.S. Pat. No., U.S. Pat.No. 8317990, U.S. Pat.No. 831895, U.S. Pat.No. 8313698, U.S. Pat. No. 6, U.S. Pat.No. 8208146, U.S. Pat.No. 8202686, U.S. Pat.No. 8137917, U.S. Pat. , U.S. Patent No. 7998436, U.S. Patent No. 7943030, U.S. Patent No. 7939021, U.S. Pat. U.S. Patent No. 7815871, U.S. Patent No. 7763471, U.S. Patent No. 7727723, U.S. Pat. , Incorporated herein by reference.

  Thus, in various embodiments, the fluid treatment panel 354 includes individual pathways, including pathways, for fluid droplets that are suitable for the assay or other process (s) performed in the reaction module 240. A grid of electrodes is formed that defines and defines a processing area. In general, “spots” or “locations” or “pads” (often referred to as “electrowetting pads” or “EWP”) are generally illustrated as rectangles in the figure, and the lines forming the sides of the rectangle are Representing the electrodes, the droplets are allowed to move along the path in separate steps from pad to pad. By manipulating the electrode grid, the droplets can be selectively moved in any of the four directions, front, back, left, or right, as needed, relative to the current position. it can. Thus, in various embodiments, the fluid treatment panel 354 includes a grid of etched electrodes that form a network of pads for moving sample droplets from sample preparation to detection of the analyte of interest. .

  In the illustrated embodiment, the electrodes formed on the substrate 356 of the fluid treatment panel 354 define a plurality of individual functional areas that are provided for the movement and / or collection of fluid droplets. As shown in FIGS. 26, 27, and 59, these areas are spaced in the sample bead area 368 spatially corresponding to the sample compartment 266 of the upper plate 241 and in the detection buffer compartment 280 of the upper plate 241. Hybridization zone 370 corresponding to the rehydration buffer zone 372 spatially corresponding to the rehydration (elution) buffer compartment 276 of the upper plate 241 and the second buffer compartment 300 of the upper plate 241. , An exonuclease reagent area 374 that corresponds spatially to the buffer plate 296 and a PCR reagent area 376 that corresponds spatially to the buffer compartment 296 of the top plate 241. Other areas defined on the fluid treatment panel 354 spatially correspond to the detection compartments 350a, 350b, 350c, and 350d, respectively, and further include electronic sensor arrays 363a, 363b, 363c, and 363d, respectively. Includes electronic sensor areas 360a, 360b, 360c, 360d. Still other paths defined on the fluid treatment panel 354 are thermal cycles, each positioned spatially below and between two adjacent bubble traps 340 on the top plate 241, or Includes PCR, pathways 364a, 364b, 364c, and 364d.

  The electrode of the fluid treatment panel 354 can further define an exonuclease area 384.

  The electrodes of the fluid treatment panel 354 may further define a detection mixing area that, in the illustrated embodiment, includes four groups of nine electrode pads indicated by reference numerals 385a, 385b, 385c, and 385d. .

  The fluid treatment panel includes a plurality of connector pad arrays configured to make contact and electrical connection with connector pins (eg, pogo pins) disposed within the treatment device, which will be described in further detail below. Further, it can be included. The illustrated embodiment includes seven connector pad arrays: 358a, 358b, 358c, 358d, 358e, 358f, and 358g.

  As will be appreciated by those skilled in the art, there are many electrode grid configurations that can be used with the composite cartridge 10, including but not limited to the configurations described herein. Exemplary electrowetting electrode configurations for different utilities are shown in previously incorporated US Pat. No. 8,541,176.

  In general, suitable materials for the fluid treatment panel 354 include printed circuit board materials. In various embodiments, the circuit board material is coated with a conductive layer to form a pattern of electrodes and interconnects (often referred to in the art as interconnects or leads) to enable lithography techniques, particularly photolithography. Insulating substrates (eg, substrate 356) that are processed using techniques. The insulating substrate is generally, but not always, a polymer. As is known in the art, one or more layers can be a “two-dimensional” board (eg, an “edge card connector” with all electrodes and interconnects in a plane) or a “three-dimensional” board ( The electrodes can be on one side and the interconnect can be used to make either the other side, through the board, or the electrodes are on multiple sides). Three-dimensional systems often rely on the use of drilling or etching to form holes or vias through the substrate, followed by electroplating with a metal such as copper, "I'm trying to make interconnections. The circuit board material is often provided with a foil, such as a copper foil, attached to the substrate, and additional copper is added as needed (eg, for interconnection), for example by electroplating. The copper surface may then need to be roughened, for example by etching, to allow attachment of the adhesive layer.

  In one embodiment, electrical connections from both the electrowetting electrode grid and the detection electrodes, i.e., the connector pad arrays 360a-g, are connected to pogo pins or similar connectors to make a connection from the chip to the processing equipment. Stretch through the panel to create a so-called land grid array that can be interfaced. In various embodiments, the surface of the fluid treatment panel 354 (eg, a PCB having an electrode grid) is coated with a film of material to facilitate electrowetting mechanisms and a clean transfer from pad to pad. The In various embodiments, the surface is coated with a polyimide film such as KAPTON® (eg, black or yellow KAPTON®) from DuPont that forms the dielectric layer. The surface properties of the dielectric layer are important for facilitating electrowetting and damping the voltage used to prevent electrolysis within the aqueous droplet. In addition, Kapton® or similar surfaces such as solder masks are required for electrowetting to function, making the surfaces hydrophobic, to name a few, such as Paralyene, TEFLON. It must be coated with a hydrophobic coating such as (registered trademark) (polytetrafluoroethylene), CYTOP (registered trademark) fluoropolymer.

  As will be appreciated by those skilled in the art, the form of reagent provided in reaction module 240 will depend on the reagent. Some reagents can be dried or in solid form (eg, certain buffers should be used), others can be dried and frozen, and the like. Particularly useful embodiments utilize dry reagents to which are added stabilizers such as salts, sugars, polysaccharides, polymers, or proteins such as gelatin, as will be appreciated by those skilled in the art. For example, Biomatrica manufactures commercial stabilizers for use in the present system.

  As will be appreciated by those skilled in the art, when used, the dry reagent can be rehydrated in one of two general ways. The method is either that the liquid from the sample preparation module 70 is introduced at a suitable pad (or area), or that the sample itself functions as an aqueous solvent that brings the solid reagent into solution. . For example, a suitable resuspension buffer (which may be water in some cases) may cause the top plate 241 from the sample preparation module 70 to a particular pad to rehydrate the reagent (s). The reagent droplet can then be mixed with the sample droplet.

  Alternatively, the droplet containing the analyte of interest (eg, in the elution buffer used to release the analyte of interest from the capture beads) is transported to a pad containing the dry reagent (s) and then the droplet It can be suspended within itself. One advantage of this embodiment is that the final volume of the droplets increases but does not increase significantly when the reagent droplets are mixed with the sample droplets. This can be particularly useful in situations where multiple reagent additions are required.

  The number, type and amount of different reagents will depend on the sample, the analyte of interest and the desired reaction. For example, for a nucleic acid target sequence in a standard PCR reaction, if the starting sample is DNA, the dry reagents on the board are RT-PCR buffer, PCR enzyme (eg, Taq polymerase), dNTP, PCR primer, exonuclease , Signal probe, signal buffer and detection buffer (lysis buffer, binding buffer, elution interference agent, (optional) included in sample preparation module 70, not dried on fluid treatment panel 354) A plurality of reconstitution buffers, and magnetic bead suspension). Exemplary embodiments are now generally described. However, as will be appreciated by those skilled in the art, any number of configurations of dry and liquid reagents in the sample preparation module 70 can be used.

  The compartment in the reaction module 240 formed between the fluid treatment panel 354 and the upper plate 241 is generally filled with a fluid in which the analyte analyte droplet (usually an aqueous solution) is insoluble, In general, it is less polar than the solution of droplets. As described in US Pat. No. 8,541,177, this disclosure is hereby incorporated by reference, but there are two general methods of isolating droplets on a pad, the methods being Fill the compartment with an insoluble fluid containing an insoluble liquid and an insoluble gas, or give the droplet an oil shell, for example by passing the droplet through an air / oil interface, and so on. It includes by using as.

  Particularly suitable insoluble fluids for use in the nucleic acid detection assays described herein include, but are not limited to, silicone oil, mineral oil, fluorosilicone oil, hydrocarbons, such as decane, andecane, dodecane. , Alkanes such as tridecane, tetradecane, pentadecane and hexadecane, dodecane, hexadecane and cyclohexane, aliphatic and aromatic alkanes such as hydrocarbon oil, mineral oil, paraffin oil, halogens such as fluorocarbons and perfluorocarbons (eg 3M Fluorinert liquid) Oil, as well as mixtures of the above. Examples of suitable gas-filled fluids include, without limitation, air, argon, nitrogen, carbon dioxide, oxygen, humidified air, and any inert gas. In one embodiment, the main phase is an aqueous solution and the secondary phase is air or oil, which is relatively insoluble in water. In other embodiments, the filling fluid comprises a gas that fills the space between the plates surrounding the droplets. A suitable filling fluid is a low viscosity oil such as silicone oil. Other suitable fluids are described in US Patent Application No. 60 / 736,399, entitled "Filler Fluids for Droplet-Based Microfluidics" filed November 14, 2005, the entire disclosure of which is Incorporated herein by reference. The fluid can be selected so that no significant evaporation of the droplets occurs.

  As will be appreciated by those skilled in the art, drop transfer from pad to pad, with the addition of reagents, as needed, can be used for any number of sample operations. In the case of nucleic acid manipulation for nucleic acid detection, these manipulations are generally performed by reagents such as PCR enzymes, PCR buffers, primers, exonucleases, reverse transcription (RT) enzymes, RT-PCR buffers, signal buffers, signal probes, etc. Addition.

  In various embodiments, one or more portions or sections of the pad's electrode grid path can be located within separate thermal zones, eg, for amplification, exonuclease digestion, reverse transcription, target elution, and electrochemical detection. Exposed to heat. Such thermal zones can include a detection region 378, an exonuclease region 380, and a thermal cycling (PCR) region (also referred to as a thermal zone) 382a, 382b, 382c.

  As will be appreciated by those skilled in the art, some operations such as PCR amplification require thermal cycling (primer binding, extension and denaturation) between a few different temperatures, while others For example, enzymatic processes such as the use of exonucleases and reverse transcriptases, specific temperature (s) for improved elution and / or reagent resuspension, or in the case of electrochemical detection Require uniform temperature for best results, such as binding / assay temperature. Isothermal amplification techniques and other alternatives to PCR typically require precise temperature control.

  In various embodiments, heat applied to different portions of the fluid treatment panel 354 is generated by thermal components such as resistance heaters or thermoelectric (Peltier) chips and within the processing bay of the device in which the cartridge 10 is located, It can be seen that it is outside the cartridge. Examples of such thermal components are described below.

  In one embodiment, the sample handling zone on the reaction panel 354 can optionally include sensors, for example, to monitor and control the temperature of the thermal zone, particularly when a particular temperature is desired. . These sensors can include, but are not limited to, thermocouples and resistance temperature detectors (RTDs). Alternatively, such a sensor can be external to the cartridge in the bay.

  In various embodiments for detecting nucleic acid objects, the fluid treatment panel 354 includes one or more thermal cycles or PCR or amplification paths 364a, 364b, 364c, and 364d. The fluid treatment panel 354 can include one, two, three or more pad thermal cycling paths. These can be used for individual PCR reactions (eg, one droplet moves up or down a path, or rises one path and descends the other) or complex (eg, multiple For a path, a plurality of different droplets can be moved up and down each path).

  As will be appreciated by those skilled in the art, each PCR reaction can be further complexed. That is, due to subject-specific amplification, the use of multiple primer sets within a single PCR reaction is cumbersome and thus the present invention allows multiple reactions to achieve high levels of complexation. To do. For example, to evaluate 21 different subject sequences (eg, in screening for respiratory viruses), for example, the first PCR sample droplet in the first pathway is a first set of seven primer pairs ( For example, “primer mix A])”, the second droplet in the second path captures the second set of seven primer pairs (“primer mix B”), and the third droplet in the third path. It may be desirable to perform three different reactions of seven primer sets, such as a second set of droplets capturing a third set ("Primermix C"). In some embodiments, more than one droplet can be processed in each path, and thus each path can include more than one set of primers. In some embodiments, the primers will be completely different in each set; in others, redundancy and / or internal control may be achieved within the system by adding the same set of primers to different pathways. Assembled into. The number of composites can be easily changed by software without having to modify any physical components of the system.

  In general, an amplification reaction suitable for use in the present system uses a set of primers, each of which has a blocking end that is impervious to standard exonucleases. That is, in order to simplify the detection reaction and remove the background signal, it is desirable to remove one helix of the double helix amplification product generated by the PCR reaction. Thus, by performing a first PCR reaction and then adding exonuclease, one helix of the double helix amplification product is digested, leaving only the detection helix.

  As generally illustrated in FIG. 59, the use of multiple heating zones along thermal cycling paths 364a-d allows droplets to travel through the appropriate thermal zones. As shown in FIG. 59, four thermal cycle paths 364a, 364b, 364c, and 364d are shown extending through three thermal zones 382a, 382b, and 382c. For example, thermal elements, such as resistance heaters, corresponding to thermal zones 382a, 382b, and 382c are heater elements outside the cartridge, and are used at temperatures of 95 ° C, 72 ° C, and 64 ° C for PCR thermal cycle use Can be maintained. In some embodiments, two different temperature zones (eg, about 95 ° C. for denaturation and about 60 ° C. for annealing and extension) can be used for a two-step PCR reaction. In other embodiments, three zones, two temperature configurations can be used, and the intermediate heater corresponding to the intermediate heat zone 382b controls the denaturation temperature (eg, about 95 ° C.) Additional heaters corresponding to the thermal zones 382a, 382c on both sides provide substantially the same annealing and elongation temperature (eg, about 60 ° C.). In this configuration, a two-step amplification cycle can be performed for two or more droplets in each thermal cycle path 364a-d. For example, two drops are placed in each thermal cycle path, and when one drop is in the denaturing zone 382b, the other is in one of the combined annealing and extension zones 382a or 382b. And vice versa. Each droplet can obtain amplification reagent (eg, primer cocktail) at a location such as, for example, each end of a thermal cycling path, such as a respective location 366a, 366b of thermal cycling path 364a-d. By reciprocating the droplets back and forth side by side between the denaturing and annealing / elongation zones, both can be amplified in the same time it normally takes to amplify a single droplet. In the four path configuration shown, this means that eight droplets can be amplified simultaneously rather than three.

  In various embodiments, the composite cartridge 10 of the present invention relies on the use of electrodes and electrochemical labels for the detection of analytes of interest. Generally, the electrode surface (optionally coated with a self-assembled monolayer (SAM)) within each electronic sensor array 363a, 363b, 363c, and 363d has a capture ligand that binds to the object. A second label ligand is included that also binds to the subject, and in the presence of the subject, the label ligand can be bound near the surface of the electrode and detected electronically.

Accordingly, the detection area of the fluid treatment panel 354 comprises one or more individual arrays of detection electrodes 363a, 363b, 363c, and 363d within the respective electronic sensor areas 360a, 360b, 360c, and 360d. As used herein, “electrode” refers to a configuration that can sense current or charge and convert it to a signal when connected to an electronic device. Alternatively, the electrode can be defined as a configuration that can apply a potential to and / or pass electrons to or from the species in the solution. Suitable electrodes are known in the art and include, but are not limited to, certain metals and their oxides including gold, platinum, palladium, silicon, aluminum; platinum oxide, titanium oxide, tin oxide, indium tin oxide, A metal oxide electrode containing palladium oxide, silicon oxide, aluminum oxide, molybdenum oxide (Mo ₂ 0 ₆ ), tungsten oxide (W0 ₃ ) and ruthenium oxide; and carbon (including glassy carbon electrode, graphite and carbon paste) Including. Suitable electrodes include gold, silicon, carbon, and metal oxide electrodes, with gold being particularly preferred. In a particularly useful embodiment, both the electrowetting electrode grid and the detection electrode are gold and are formed simultaneously on the fluid treatment panel 354.

  The system is particularly useful in the form of an array, ie there is a matrix of detection electrodes that can be addressed. “Array” as used herein means a plurality of capture ligands on an electrode in the form of an array, the size of the array will depend on the configuration of the array and the end use. An array can be made that includes from about two different capture ligands to about 50-100. In some preferred embodiments, 80 or 100 working detection electrodes are divided into 20 4 or 5 individual zones, each zone containing up to 60 capture probes (3 per electrode) With different capture probes).

  The detection area of the fluid treatment panel 354 includes one or more arrays of detection electrodes 363a-d, each of which is an electronic sensor area 360a- through the fluid with one associated droplet path of the detection mixing area 385a-d. d. That is, the droplets containing amplification products are, for example, in dry form, at label probes (eg, locations 362a, 362b, 362c, and 362d) adjacent to electronic sensor detection areas 360a, 360b, 360c, and 360d, respectively. A single probe cocktail that can be acquired) and then sent to the associated electronic sensor detection areas 360a, 360b, 360c, and 360d. The signal probe cocktail is applied to a portion of the top plate 241 that forms the locations 362a, 362b, 362c, and 362d or a portion of the fluid treatment panel 354 that covers the locations 362a, 362b, 362c, and 362d. Can do. In general, each detection zone receives one or more sample droplets that are typically sent over an array of electrodes and can be considered as one large “pad”.

  In one embodiment, the reaction module 240 includes four electronic sensor detection zones, each electronic sensor array including 20 working electrodes (which can include one reference electrode and one spare electrode). )including. Each sensing electrode of each electronic sensor array 363a-d includes independent leads (interconnects) to transfer input and electronic response signals to each electrode of the array, both input and electronic response signals being independently Each electrode can be monitored. That is, each electrode can be addressed independently. Furthermore, the reaction module is preferably configured for independent control of the electrowetting pad surrounding each electrode of each electronic sensor array 363a, 363b, 363c, and 363d.

  In addition to the components of the fluid treatment panel 354 described above, the fluid treatment panel 354 also optionally includes an EPROM, EEPROM, or RFID to identify the cartridge, including, for example, information about the batch, treatment, or contents of the composite cartridge 10. Can be included. This can include, for example, information about the identification of the assay.

[Device Overview]
An apparatus configured to process the composite cartridge 10 and embody aspects of the present invention is indicated by reference numeral 400 in FIG. The apparatus includes a control console 402, one or more processing modules 410 operably coupled to the control console 402, and a plurality of processing bays within each processing module 410, each processing bay having a composite cartridge. Accepted and configured to process composite cartridges independently of other bays, the device comprises device software (ISW). In various embodiments, the apparatus comprises one control console 402 and up to four processing modules 410, each processing module including six processing bays. Each processing module 410 is operably coupled to the control console 402 to exchange power, input / output data, and control signal transmission with the control console 402, for example, as well as physically to the control console 402. Can be connected. Each processing bay having a processing module 410 is configured to accept one composite cartridge 10 at a time and process the cartridge independently of other processing bays that process other composite cartridges. In various embodiments, the apparatus is configured such that each processing bay completes cartridge processing in 60 minutes or less.

  The ISW provides a graphical user interface for the user to initiate processing, receive results, and provide inputs that at least partially control the operation of the device. In various embodiments, the ISW is configured to run on a Windows® computer having a touch screen 404 located on a control console 402 that provides the primary functionality for user input. In various embodiments, the apparatus is configured to provide a connection to a local area network (“LAN”) and a laboratory information system (“LIS”). The device may also include a barcode scanner (not shown) that facilitates logging into the ISW, sample tracking, and positive ID features of the device.

  The control console 402 of the device includes a touch screen panel 404, a system computer, a power source, a connection to an external data system, and a connection to the processing module (s) and a processing bay. In various embodiments, a power source at the control console provides power to the entire device. A cable from the control console provides power transmission and provides data transfer to and from the processing bay. In various embodiments, the control console also includes equipment for physically attaching one or more processing modules to the control console.

  Each processing bay includes hardware, firmware, and electronics that perform the assay on the composite cartridge 10. Each processing bay may include a bay PCB. In various embodiments, the bay PCB can handle the processing bay electronics and firmware (such as the microprocessor and firmware on the microprocessor) and power in the composite cartridge (eg, up to 300V on the electrowetting pad). A circuit for supplying, a circuit for performing electronic detection of a reaction product on the composite cartridge, a circuit for controlling a heater in a processing bay that interacts with the composite cartridge, and a circuit for measuring and controlling the temperature in the composite cartridge And circuitry for controlling the movement of the various moving components of the processing bay and circuitry for controlling the pump of the processing bay.

  Each processing bay can also include a connector PCB. In various embodiments, the connector PCB contacts the composite cartridge and has a pogo pin configured to transfer data, control signals, and power between the composite cartridge and the processing bay PCB, and within the processing bay. A pogo pin configured to be in electrical contact with the heater element.

  Each processing bay further includes a stepper motor. In various embodiments, the processing bay includes two stepper motors. One stepper motor controls the placement of magnets, heaters, and pogo pins or other connector elements relative to the composite cartridge, and one stepper motor compresses the blister on the composite cartridge and pre-determines the blister. The sequence controls the cam follower plate in the processing bay that discharges the contents.

  Each processing bay also compresses the blisters of the composite cartridge 10 in a specified sequence and activates the active valve of the composite cartridge 10, thereby discharging the contents of the cartridge blisters in the specified sequence. A blister compression assembly configured as described above. In various embodiments, the blister compression mechanism assembly comprises a blister compression actuator or an array of compression mechanisms, each comprising a cam arm configured to press a compression pad over the blister. The blister compression mechanism assembly includes a cam arm plate on which a cam arm and a compression pad of the compression mechanism are operatively mounted on a blister for movement between a retracted position and an extended blister compression position; Cam arm to actuate the cam arm to compress the blister in a sequence that is movable with respect to the cam arm plate and determined by the relative positions of the compression mechanism in the groove and ridge of the cam arm plate and cam follower plate And a cam follower plate that includes grooves with ridges (or other cam follower elements) that are arranged and sequenced to engage the cam arms of the actuator array as the cam follower plate moves relative to the plate.

  Each processing bay may also include a pump coupled to the composite cartridge 10 via a pump port 104 and configured to provide an accelerating force for reagents and samples in the sample preparation module of the composite cartridge.

  Each processing bay also has an LED PCB 466 (FIG. 38) that provides an LED indicator of the processing bay status and a light sensor that detects the condition in the composite cartridge, for example, via the entrance light port 14 and the exit light port 16. To FIG. 41).

  Each processing bay also includes on-board hardware configured to mount the processing bay within the processing module and an electrical connector configured to transfer power and data between the processing bay and the processing module. be able to.

  Each processing bay also heats the upper bay or cartridge carriage assembly with the blister compression mechanism assembly and the composite cartridge processing assembly, the lower bay with the heating and control assembly, and the composite cartridge held in the cartridge carriage assembly. May include a composite cartridge carrier configured to provide physical connection and alignment with a cam frame assembly configured to move the control assembly;

[Control console]
A processing apparatus embodying aspects of the present invention and configured to process the composite cartridge 10 described above is indicated by reference numeral 400 in FIG. As described above, the apparatus 400 includes a control console 402 and one or more processing modules 410 operatively associated with the control console 402. The control console 402, in one embodiment, presents a graphical user interface and includes a touch screen that allows a user to enter information into the control console 402 and / or information can be presented to the user. A display panel 404 is included. In various embodiments, the control console 402 can include additional or alternative means for entering data, such as a keyboard, microphone, switch, manual scanner, voice sensitive input, and the like. As further noted above, this apparatus can be used with other types of machine readable codes, such as barcode scanners for reading barcodes such as one-dimensional or two-dimensional barcodes, or RFID scanners. A scanner can be included. In various embodiments, the control console 402 may include a hard drive or other storage medium, a monitor, a printer, an indicator light, or an acoustic signal element (eg, buzzer, horn, bell, etc.), email, text message, etc. Additional or alternative means for outputting data (ie information and / or results) can be provided.

[Process module]
As shown in FIGS. 32 and 34, each processing module 410 includes one or more cartridge doors 412, each cartridge door 412 having a processing bay (hereinafter described) in which the cartridge 10 can be processed. Are described). In the illustrated embodiment, each processing module 410 includes six cartridge doors 412 and associated processing bays. Each cartridge door 412 is configured to receive the composite cartridge 10 preferably in a single preferred orientation. Each cartridge door is also preferably a closeable door (eg, a pivoting door panel) that is biased in the closed position, eg, by a spring, but that can be opened when the cartridge is inserted therein. Includes.

  In various embodiments, each processing module 410 is operatively coupled to the control console 402. The processing module 410 can be electrically coupled to the control console 402 to allow electronic communication between the control console 402 and the processing module 410. Such electronic communications can include power transfer from the control console to the processing module to power various electronic components within the processing module, control signals, input data, output data, and the like.

  Each processing module 410 can also be physically connected to the control console 402 adjacent, for example as shown in FIG. As in the illustrated embodiment, the apparatus 400 can include one or more processing modules 410 secured to one or both sides of the control console 402. Additional processing modules can be secured to other processing modules adjacent to one or both sides of the control console 402. In one preferred arrangement, the apparatus 400 includes up to two processing modules 410 secured to each side of the control console 402, each processing module 410 having six cartridge doors 412 and six per processing module. And an associated processing bay for processing up to the composite cartridge 10.

  The control console 402 and the processing module 410 are provided in a modular format as shown, for example, adding one or more processing modules 410 or subtracting one or more processing modules 410 from a single control console 402. This facilitates device scalability and facilitates device troubleshooting so that a processing module 410 having one or more malfunctioning processing bays can be removed from the device for repair or replacement. Preferably, the device is still usable with the remaining operable processing module 410.

  In another embodiment, however, the input screen associated with the control console-and / or other input means-and one or more-preferably-multiple cartridge doors and associated processing bays comprise a single enclosure. Having a single, integrated device.

  Further details of the processing module 410 are shown in FIGS. 35, 36, and 37. Each processing module 410 includes a plurality of cartridge doors 412 and associated processing bays 440. The illustrated embodiment includes six processing bays 440. The processing bays 440 are arranged in a stacked configuration within the housing of the processing module 410. Each processing bay 440 has a frame 418 that partially surrounds the processing bay with a horizontal top panel 420 and a vertical back panel 422 associated therewith (see FIG. 37). As shown in the figure, the front panel 413 of the processing module 410 in which the cartridge door 412 is disposed is oriented at an inclined angle from the lower part of the processing module 410 to the upper part of the processing module 410. This may be due to ergonomic and / or aesthetic reasons. In other embodiments, the front panel of the processing module can be vertical. Due to the angle of the front panel 413 of the processing module 410, each processing bay 440 is offset horizontally (i.e., backward) with respect to the processing bay immediately below.

  In various embodiments, each processing bay 440 is associated with a ventilation fan 416 secured to the vertical panel 4222 of the housing 418 and a ventilation duct extending from the ventilation fan 416 to the back wall of the housing of the processing module 410. 414. As shown in the figure, the ventilation duct 414 shortens the length from the lowermost processing bay 440 to the uppermost processing bay due to the inclination of the front panel 413 and the horizontal displacement of the processing bay 440.

  The processing module 410 can further include additional structural elements for securing each of the processing bays 440 within the processing module housing. Processing bays 440 and processing modules 410 preferably each bay 440 is independent of processing module 410 if one or more processing bays 440 are malfunctioning or otherwise require maintenance or repair. Configured so that it can be removed and replaced to facilitate equipment repair.

[Processing bay]
Processing bay 440 is shown in the various views of FIGS. 38, 39, 40, and 41. In each of FIGS. 38 to 41, the frame 418 of the processing bay 440 is omitted from the drawings. FIG. 38 is a front, right side perspective view of the processing module 440 with the composite cartridge 10 inserted therein. FIG. 39 is a front and left perspective view of the processing module 440 in which the composite cartridge 10 is inserted. 40 is a rear perspective view of the processing module 440. FIG. FIG. 41 is an exploded perspective view of the front and right side of the processing module 440 in which the composite cartridge 10 is inserted.

  Each processing bay 440 is configured and arranged to form a lower floor of the processing bay 440, including fluid leakage that may arise from the composite cartridge 10, and to provide support and mounting structure for the various components of the processing bay 440. A drip tray 446. The main PCB (printed circuit board) 442, also referred to as a bay PCB, provides the main control of the processing bay 440 along with data and power distribution and transmission. Flexible connector 444 connects bay PCB 442 to connector PCB (described below, not shown in FIGS. 38-41) within processing bay 440, as will be described in more detail below. The processing bay 440 receives two tubular female alignment elements 460 that receive male alignment elements disposed within the processing module 410 to properly align and position the processing bay 440 at the bay mounting location within the processing module 410. , 462, and the like.

  Processing bay 440 is along a functional line between cartridge processing assembly 470 (also known as lower bay) and blister (or deformable chamber) compression mechanism assembly 750 (also known as upper bay). Can be divided conceptually. The main functions of the cartridge processing assembly 470 are to accept the cartridge 10, secure the cartridge in the bay 440, apply heat and magnetic force to the processing module 240 of the composite cartridge 10, apply control power to the composite cartridge 10, and Ten rotary mixers 192 are engaged, power is applied to the rotation of the rotary mixer 192, and the cartridge 10 is ejected from the processing bay 440 at the end of an assay or other process performed in the bay 440. The primary function of the blister compression mechanism assembly 750 is to rupture the various deformable chambers of the composite cartridge 10 in an appropriate sequence. Each of these various components will be discussed in more detail below.

  Processing bay 440 is an LED for controlling one or more LEDs that provide information to the user, such as indicating the status of processing bay 440 and / or indicating whether a cartridge is positioned in processing bay 440. A PCB 466 is further included. The status LED is visible on the front panel 413 of the processing module 410 via a light pipe or other light transmitter that provides a light indication signal adjacent to the cartridge door 412 associated with the bay 440. The LED PCB 466 also detects fluid flowing through the inlet and outlet light detection chambers 154 and 158 of the sample preparation module 70 via the inlet and outlet light ports 14, 16 (eg, generates a signal). It is possible to control the optical sensor configured and arranged as described above.

  The side walls 472, 474 may extend upward along opposite sides of the processing bay 440 and be secured to elements 443, 445 that extend above the drip tray 446. The mounting plate 640 includes a generally horizontal blister plate 644 (see FIG. 42) that is secured to the upper ends of the side walls 472, 474 and generally separates the cartridge processing assembly 470 from the blister compression assembly 750.

  In various embodiments, each processing bay 440 further includes an encoder 838 and a cam frame motor 602 associated with the cam follower motor 834. The cam plate motor 834 and the cam frame motor 602 are fixed to the motor mount 642 of the mounting plate 640 (see FIG. 42).

  Pump 458 provides pressure applied to composite cartridge 10 via pump port 104.

  As described in more detail below, cartridge processing assembly 470 includes a Peltier heater assembly for generating a thermal process in processing bay 440. In order to vent the process bay 440 and dissipate excess heat generated by the Peltier heater, the process bay 440 can include a Peltier ventilation assembly. The ventilation assembly includes a cooling fan 448 attached to the fan mount 450 of the drip tray 446 and disposed in front of the airflow duct extending between the cooling fan 448 and the Peltier heating assembly in the process bay 440. In various embodiments, the airflow duct can include a cooling duct 452 and a duct cover 456 extending between the cooling fan 448 and the starting position of the cooling duct 452 (see FIGS. 39 and 41).

Cartridge handling assembly (lower bay)
An embodiment of the cartridge processing assembly 470 is shown in FIGS. As noted above, most features of the cartridge processing assembly 470 are located below the blister plate 644 of the motor mount 642. The cartridge processing assembly 470 includes a cartridge carriage assembly 650 configured to receive, hold, and later eject the composite cartridge 10. The cartridge carriage assembly 650 is fixed to the lower surface of the blister plate 644 of the mounting plate 640.

  The cam block assembly 600 surrounds the cartridge carriage assembly 650 on three sides and is supported on linear cam followers 480a, 480b that extend from each of the side walls 472, 474 to a follower slot 612 formed on each side of the cam frame 606. A cam frame 606 mounted for linear back-and-forth movement in the processing bay 440.

  The mixing motor assembly 700 is pivotally connected to the blister plate 644 below the blister plate and operatively engages and disengages the rotary mixer 192 of the composite cartridge 10 disposed within the cartridge carriage assembly 650. It is configured to be attached in a pivot shape.

  The heating and control assembly 500 is disposed under the cartridge carriage assembly 650 so that when the cartridge is inserted into the cartridge carriage assembly 650, the heating and control assembly 500 is selectively brought into contact with the bottom surface of the composite cartridge 10. Operatively coupled to the cam frame 606 and the cam block assembly 600 for converting longitudinal movement of the longitudinal cam frame 606 into vertical movement of the heating and control assembly 500.

[Cartridge carriage assembly]
Further details of the cartridge carriage assembly 650 are shown in FIG. 46, which is an exploded perspective view of the cartridge carriage assembly 650 with the other components of the cartridge processing assembly 470 omitted. The cartridge carriage assembly 650 includes a carriage holder 652 with a generally rectangular frame that is secured to the lower portion of the blister plate 644 of the mounting plate 640. A detector can be provided to detect when the composite cartridge 10 (not shown in FIG. 46) is inserted into the cartridge holder 652. In various embodiments, the detector comprises a photodetector comprising a light emitter 686 and a detector 688, each disposed in a respective pocket on the opposite side of the cartridge holder 652. The light beam from the emitter 686 to the detector 688 is blocked when the composite cartridge is inserted into the cartridge holder 652, thereby generating a signal indicating the presence of the cartridge.

  Cartridge latch 654 is mounted for pivotal movement at the closed end of cartridge holder 652. Cartridge latch 654 is pivotally mounted on latch pin 660 for rotation about a horizontal axis of rotation. The cartridge latch 654 further includes a front hook 656 and a rear lever 658. The torsion spring 662 biases the latch 654 in the rotational direction so that the hook 656 is in the upper position. When the cartridge 10 is inserted into the cartridge holder 652, the cartridge pushes down on the hook until the hook 656 of the cartridge latch 654 engages the recess in the lower portion of the cartridge 10 and the lower portion of the flash 30. The bias of the torsion spring 662 holds the hook 656 in its recess to hold the cartridge in the cartridge holder 652.

  The cartridge discharge assembly 670 includes a discharge rack 672 disposed in a discharge bracket 682 extending from the rear end of the cartridge holder 652. The linear gear teeth of the discharge rack 672 are coupled to the rotary damper 676 and mesh with a damper pinion gear 674 mounted for rotation on the discharge bracket 682 adjacent to the discharge rack 672. The spring catch pin 680 extends through the discharge rack 672 and is supported at its end by the end wall of the discharge bracket 682. The compression spring 678 is disposed between the end of the discharge rack 672 and the end of the spring catching pin 680. Accordingly, the discharge rack 672 is biased in the longitudinal direction toward the open end of the cartridge holder 652. A limit stop element can be provided to prevent the cartridge rack 672 from being pushed too far by the spring 678. The discharge rack 672 initially extends into the cartridge holder 652 and is contacted by the end of the composite cartridge 10 inserted into the cartridge holder 652. As the cartridge is further inserted into the cartridge holder 652, the discharge rack 672 is pushed back, thereby compressing the spring 678 and processing the cartridge 10 longitudinally toward the open end of the cartridge holder 652. A bias force is generated that pushes out of bay 440. Since the cartridge latch 654 captures the fully inserted composite cartridge, the ejection assembly 670 prevents the cartridge from being pushed back out of the cartridge holder 652.

  The cartridge latch switch 666 is located at the closed end of the cartridge holder 652 and emits a signal when the composite cartridge is inserted into position within the cartridge holder 652 so that the cartridge will engage with the cartridge latch 654. It is configured. Upon completion of an assay or other process performed in the processing bay 440, the cartridge latch 654 is pivoted against the bias of the torsion spring 662, as described below (in the illustrated embodiment, counterclockwise). Thereby releasing the composite cartridge held in the cartridge holder 652. Upon opening of the cartridge, the cartridge is discharged by the energy stored in the compression spring 678 that bears against the discharge rack 672. An operatively associated rotary damper 676 that engages the damper pinion 674 and the discharge rack 672 ensures a controlled release of the discharge rack 672 and prevents the composite cartridge 10 from being suddenly discharged from the cartridge holder 652.

[Heating and control assembly]
Details of the heating and control assembly 500 are shown in FIG. 43, which is an exploded perspective view of the heating and control assembly 500 with the other components of the cartridge processing assembly 470 omitted.

  The heating and control assembly 500 includes a support plate 502, a connector PCB 504 supported on the support plate 502, a cover plate 550 partially covering the connector PCB 504, a cartridge magnet assembly 552, a sample preparation magnet assembly 570, And a magnet actuator 584 disposed under the support plate 502. Front alignment pins 416 and back alignment pins 414 extend upward from support plate 502.

  The air connector 518 is attached to the air ports 519a, 519b of the cover plate 550. The air connector 518 provides a connection between a pressure source, such as, for example, a pump 458 and the cartridge 10 via the pump port 104 and between the external valve in the processing bay 440 and the passive valve assembly 220 of the cartridge. Is provided via a passive valve port 108 (see FIG. 15).

  43 and 44, which are top plan views of the connector PCB 504, the connector PCB 504 includes an elution heater assembly 506, a detection Peltier assembly 540, and PCR heater assemblies 520a, 520b, and 520c. In various embodiments, the elution heater assembly 506 was comprised of a resistive heating element attached to a dedicated PCB and a thermally conductive material attached to the resistive heating element or otherwise thermally coupled. A heat spreader. Similarly, in various embodiments, each element 520a, 520b, and 520c of the PCR heater assembly is attached to a resistive heating element attached to a dedicated PCB, to a resistive heating element, or in other cases a thermal heating element. And a heat spreader made of a thermally conductive material that are coupled to each other.

  Details of the detection Peltier assembly 540 are shown in FIG. 45, which is an exploded perspective view of the Peltier assembly 540. Assembly 540 includes a Peltier device 544 (ie, a thermoelectric element) that is coupled to a power source and control printed circuit board 546. The heat spreader 542 is preferably composed of a thermally conductive material, but is disposed on top of the Peltier device 544. The heat sink 548 is disposed under the Peltier chip 544. The heat sink 548 can comprise a panel that is in surface contact with the surface of the Peltier device 544 and then has a plurality of heat dissipating rods (or fins) that are stretched and formed of a thermally conductive material. The detection Peltier assembly 540 is mounted in an associated opening formed in the support plate 542 and at least a portion of the heat sink 548 extends therethrough. The heat dissipating rod of the heat sink 548 extends below the support plate 502 and is located at the end of the Peltier cooling duct 452 (see FIGS. 39 and 41). In one embodiment, the detection Peltier is configured to apply a thermal gradient, for example, to reduce the temperature of the detection region, eg, the detection region 378 of the composite cartridge 10.

  A plurality of connector pin arrays 510a, 510b, 510c, 510d, 510e, 510f and 510g are disposed around the connector PCB 504 and between the connection pads of the associated connector pad arrays 358a-358g of the fluid treatment panel 354 of the composite cartridge 10. It comprises an array of connector pogo pins that come into contact and enable electrical connection (see FIG. 58). The connection between the connector pin arrays 510a-510g and the connector pad arrays 358a-358g provides a connection between the device 400 and the composite cartridge 10, for example for power, control signals, and data. For example, the connection between the connector pin arrays 510a-510g and the connector pad arrays 358a-358g can be performed from the device to an electrowetting grid (eg, thermal cycling tracks 364a-364d, sample bead area 368, hybridization area 370, elution buffer. Provides power and control to the agent zone 372, exonuclease reagent zone 374, PCR reagent zone 376, detection mixing zone 385a-385d, and exonuclease zone 384). In addition, the connection between the connector pin arrays 510a-510g and the connector pad arrays 358a-358g provides power to and receives data from the electronic sensor arrays 363a-363d.

  As shown in FIG. 43, the connector PCB 504 further includes a plurality of heater pins 512- which may comprise pogo pins, which connect to the various heater assemblies 540, 506 and 520a, b, c.

  The heating and control assembly 500 further includes a cartridge magnet assembly 522 and a sample preparation magnet assembly 570.

  Details of the sample preparation magnet assembly 570 are shown in FIG. 49A, which is a top perspective view of the sample preparation magnet assembly. The sample preparation magnet assembly 570 includes a magnet holder 572 mounted on a horizontal spindle 574 so that the support plate 502 can rotate about the spindle 574. A torsion spring 576 biases the sample preparation magnet assembly 570 downward. The actuator bracket 578 extends below the magnet holder 572, and the magnet 580 is supported on top of the magnet holder 572 and is secured thereto, for example, with a suitable adhesive. When deployed upward and rotated with respect to the bias of the torsion spring 576, the magnet 580 extends through aligned openings formed in the support plate 502, connector PCB 504, and cover plate 550.

  When deployed, the sample preparation magnet assembly 570 is disposed adjacent to the capture chamber 100 of the sample preparation module 70 of the composite cartridge 10, thereby energizing the fluid contained within and flowing through the capture chamber. Apply.

  Details of the cartridge magnet assembly 552 are shown in FIG. 49B, which is a top perspective view of the cartridge magnet assembly. The cartridge magnet assembly 552 includes a magnet holder frame 554 and a magnet array 556 disposed within the magnet holder frame 554. The magnet array 556 can comprise individual magnets (eg, three) and can be surrounded from four directions by a magnet holder frame 554 to form a frame surrounding the magnet array 556. The magnet array 556 can be secured within the magnet holder frame 554 by, for example, a suitable adhesive. The converging magnet 558 is disposed in the opening of the upper part of the frame of the magnet holder 554. In one embodiment, the focusing magnet 558 is cylindrical and can comprise neodymium N52. The converging magnet 558 converges the magnetic force of the magnet array 556 to a relatively small area in order to attract the magnet target capturing beads to a small area. The magnet holder 554 is mounted on a horizontal spindle 560 that is connected to the support plate 502, allowing the magnet holder 554 and the magnet array 556 to rotate around the spindle 560. Torsion spring 562 biases cartridge magnet assembly 552 downward. Actuator bracket 566 extends below magnet holder 554. The magnet holder 554 is rotated upwardly against the bias of the torsion spring 562 and the top of the magnet assembly 552 has aligned openings formed in the support plate 502, connector PCB 504, and cover plate 550. Stretch through.

  When deployed, the cartridge magnet assembly 552 is positioned adjacent to the sample chamber 266 of the reaction module 240 adjacent to the location indicated by reference numeral 270 (see FIG. 26).

  Returning to FIG. 43, the cam followers 590 a and 590 b extend from the opposite side of the support plate 502, and the slot follower 592 extends from the opposite side of the support plate 502. The slot follower 592 extends into a slot 476 formed in each of the side walls 472, 474 (see FIG. 42) and is vertically movable therein, with the vertical movement of the support plate 502 relative to the side walls 472, 474. The support plate 502 is prevented from moving horizontally with respect to the side walls 472 and 474.

[Cam frame assembly]
Details of the cam frame assembly 600 are shown in FIG. 47, which is an exploded perspective view of the cam frame assembly 600 with the other components of the cartridge processing assembly 470 omitted. Cam frame assembly 600 includes a cam frame motor 602 that drives a linear actuator 604. The cam frame 606 is opposed to a generally parallel, elongated circular material 608, 610, and a cross circular material 614 extending between corresponding ends of the elongated circular materials 608, 610, including. The linear actuator 604 is coupled to the cam frame 606 at a motor connector 618 that projects upward from the cross disc 614. A follower slot or channel 612 is formed along the exterior side below the top surface of each of the elongated circular members 608, 610. Follower elements 480a, 480b extending from each of the side walls 472, 474 extend into the follower slot 612 (see FIG. 42).

  The cam rail 620a is fixed to the long circular member 608, and the cam rail 620b is fixed to the long circular member 610. The upper edge of the cam rail 620a cooperates with a follower slot 612 formed at the lower outer end of the elongate circular member 608 to form a channel that receives the cam followers 480a, 480b, The cam rails 620a and 620b can be moved in the longitudinal direction with respect to the side walls 472 and 474, and the cam frame 606 is prevented from moving in the vertical direction with respect to the side walls 472 and 474.

  Each cam rail 620a and 620b includes a front cam slot 622a and a rear cam slot 622b. Cam followers 590a, 590b projecting from the sides of the support plate 502 of the heating and control assembly 500 (see FIG. 43) extend into the cam slots 622a, 622b, respectively. Each cam slot 622a, 622b has a lower horizontal segment (right segment in FIG. 47), an upper horizontal segment (left segment in FIG. 47), and an angled transition between the lower and upper horizontal segments. Have.

  Before the composite cartridge 10 is inserted into the cartridge carriage assembly 650, the cam frame 606 is in a relatively forward position relative to the heating and control assembly 500, and the cam followers 590a, 590b extending from the support plate 502 are cam slots 622a. , 622b in the lower horizontal segment (the right segment shown in FIG. 47). Accordingly, the support plate 502 and the entire heating and control assembly 500 are in a lower position relative to the cartridge carriage assembly 650. When the composite cartridge 10 is inserted into the cartridge carriage assembly 650, the alignment fork 246 (see FIG. 24) of the top plate 241 engages the back alignment pin 514—which is longer than the front alignment pin 516 and the support plate 502 is below The cartridge carriage assembly 650 is extended to the proper position in the cartridge assembly 650.

  As shown, after the composite cartridge is inserted into the cartridge carriage assembly 650, for example, when the cartridge latch switch 666 is triggered by the end of the fully inserted cartridge, the cam frame motor 602 is linear. Actuated to retract the actuator 604 and the cam frame 606 attached thereto. This causes movement of the cam rails 620a, 620b relative to the support plate 502, thereby moving the cam followers 590a, 590b from the bottom of the cam slots 622a, 622b, the right horizontal segment, up an angled transition, The top of the cam slots 622a, 622b is moved to the left horizontal segment, thereby raising the support plate 502 and the heating and control assembly 500 into contact with the composite cartridge disposed within the cartridge carriage assembly 650.

  For the cartridge held in the cartridge carriage assembly 650, the support plate 502 is raised so that the front alignment pins 516 of the support plate 502 extend into an alignment loop 244 that extends from the top plate 241 (see FIG. 24). ). The cartridge is substantially secured within the cartridge carriage assembly 650 by rear alignment pins 514 that are engaged by alignment forks 246 and front alignment pins 516 that extend into alignment loops 244.

  Raising the heating and control assembly 500 relative to the cartridge 10 held within the cartridge carriage assembly 650 causes the connector pin array 510a-510g of the connector PCB 504 to be moved into the respective connector pad array of the fluid treatment panel 354 of the composite cartridge 10. It arrange | positions so that 358a-358g may be contacted. Furthermore, the elution heater assembly 506 of the connector PCB 504 is in contact with (or to allow transfer of thermal energy) a portion of the fluid treatment panel 354 corresponding to the exonuclease region 380 or in the vicinity thereof. Brought. Similarly, the components of PCR heater assembly 520a, 520b, 520c of connector PCB 504 are in contact with a portion of fluid treatment panel 354 corresponding to thermal cycling regions 382a, 382b, and 382c (ie, providing thermal energy transfer). To make it possible) or in the vicinity. The sensing Peltier assembly 540 of the connector PCB 504 is brought into contact with (or to allow heat energy transfer) a portion of the fluid treatment panel 354 corresponding to the sensing region 378 or in the vicinity thereof. . The air connector 518 is also brought into contact with the pump port 104 and the passive valve port 108 of the sample preparation module 70 of the composite cartridge 10.

  Each cam rail 620a, 620b is connected to each cam frame 606 by two threaded spring capture posts 624a, 624b having compression springs 626a, 626b disposed between the cam rail 620a and the respective heads of the posts 624a, 624b. It is fixed to the circular members 608 and 610 in the longitudinal direction. This “shock absorbing” configuration allows for a certain amount of movement of the cam rails 620a, 620b relative to the elongate circular members 608, 610 so that the heating and control assembly 500 can be configured with Prevents being pushed against the bottom. Accordingly, the heating and control assembly 500 will be pushed against the bottom of the composite cartridge with a force equivalent to the compression force of the springs 626a, 626b.

  43 and 48, which are cross-sectional perspective views of the cam frame and magnet actuator 584 of the cartridge processing assembly 470, the magnet actuator 584 is coupled to the cam frame 606 so that the cartridge is within the cartridge carriage assembly. When inserted, the cartridge magnet assembly 552 and the sample preparation magnet assembly 570 are configured to rotate relative to the composite cartridge to an operable position. The magnet actuator 584 includes a spring 587 that biases the actuator to the left in FIG. Magnet actuator 584 includes a vertical tab 585 configured to mate with actuator bracket 566 of cartridge magnet assembly 552 and a vertical tab 586 configured to mate with actuator bracket 578 of sample preparation magnet assembly 570. The magnet actuator 584 is coupled to the cam frame 606 by a magnet actuator hook 628 that extends below the crossbar 614 and engages a hook loop 589 formed at the end of the magnet actuator 584.

  As described above, the cam frame 606 is in the forward position before the composite cartridge 10 is inserted into the cartridge carriage assembly 650. The magnet actuator 584 is biased forward (left) by a spring 587, and the cartridge magnet assembly 552 and the sample preparation magnet assembly 570 are each retracted into the retracted position by the force of their respective torsion springs 562, 576. Rotated in the direction. In this context, the retracted positions of the cartridge magnet assembly 552 and the sample preparation magnet assembly 570 are positioned such that the cartridge magnet assembly 552 and the sample preparation magnet assembly 570 do not apply a large magnetic force to any part of the composite cartridge 10. . After the composite cartridge is inserted into the cartridge carriage assembly 650, the cam frame 606 is retracted by the cam frame motor 602 and the linear actuator 604 as described above (to the right of FIG. 48). As the cam followers 590a, 590b of the support plate 502 move from the lower, right horizontal segment of the cam slot 622a, 622b, up an angled transition to the upper, left side horizontal segment of the cam slot 622a, 622b. In addition, retraction of the cam frame 606 causes the heating and control assembly 500 to be raised to contact the composite cartridge 10.

  The magnet actuator 584 coupled to the cam frame 606 by the magnet actuator hook 628 also moves with the cam frame 606 and pulls the magnet actuator 584 to the right of FIG. When the actuator bracket 584 is pulled by the moving cam frame 606, the vertical tab 585 that engages the actuator bracket 566 of the cartridge magnet assembly 552 will rotate upward, counterclockwise, as shown in FIG. The magnet assembly 552 is rotated. Similarly, the vertical tab 586 of the actuator bracket 584 that engages the actuator bracket 578 of the cartridge magnet assembly 570 rotates the magnet assembly 570 in an upward direction, counterclockwise, as shown in FIG. . Depending on the length of the upper horizontal segment of each of the cam slots 622a, 622b, the cam frame 606 and the cam rails 620a, 620b can move in the length direction relative to the support plate 502, and the cam followers 590a, 590b The composite cartridge disposed in the cartridge carriage assembly 650 is disposed in the upper horizontal segment without changing the height positions of the support plate 502 and the heating and control assembly 500. In various embodiments, the magnet actuator bracket 584 is configured relative to the cartridge magnet assembly 552 and the sample preparation magnet assembly 570 so that the cam frame 606 raises the support plate 502 and the heating and control assembly 500 (right When the support plate 502 and the heating and control assembly 500 are first raised to contact the composite cartridge (ie, the cam followers 590a, 590b of the support plate 502 are initially moved to the cam slots 622a, 622b). ), The magnet assemblies 552, 570 are not initially deployed (or not fully deployed). Further (rightward) movement of the cam frame 606 (the longitudinal length of the upper horizontal segment of the cam slots 622a, 622b supports the cartridge carriage assembly 650 and the composite cartridge held therein. Will not change the position of the plate 502 and the heating and control assembly 500), further pulling the magnet actuator bracket 584 into contact with or near the composite cartridge to place the magnet assembly 552, 570 in a fully deployed position ( Full rotation (counterclockwise). Thus, with the support plate 502 and the heating and control assembly 500 maintained in an upper position in contact with the composite cartridge, the magnet assembly is configured for movement independent of the rest of the heating and control assembly 500, and the cam frame 606 includes To enable selective deployment of the magnet assemblies 552, 570 to assist in the requirement to selectively apply or remove magnetic force to the composite cartridge held within the cartridge carriage assembly 650. , Can move in the long direction.

  Also, as best seen in FIG. 48, a linear actuator that extends over the crossbar 614 as the cam frame 606 is advanced relative to the cartridge to lower the heating and control assembly 500 (to the left in FIG. 48). The connector 618 contacts the lever 658 of the cartridge latch 654, thereby rotating the cartridge latch 654 counterclockwise to lower the hook 656 and remove the hook 656 from the composite cartridge, and the composite cartridge is removed from the cartridge ejection assembly 670. Thus, the cartridge holder 652 can be discharged.

[Mixed motor assembly]
Details of the mixing motor assembly 700 are shown in FIGS. 50A and 50B. FIG. 50A is a perspective view of the mixing motor assembly 700, and FIG. 50B is an exploded perspective view of the mixing motor assembly 700.

  The mixing motor assembly 700 includes a mixing motor bracket 702 on which a mixing motor 706 is mounted. Suitable motors include Polol Micro Metal Gearmotor with 150: 1 gearbox and Maxon, model DCX10L EB SL 4.5V with 64: 1 gearbox. Suitable motor characteristics include a 12 oz-in torque, 100 rep, 45 ° C. operating environment, a 3000 hour life, and a compact size (eg, less than 10 mm wide (diameter) and 25 mm long).

  The bevel gear 708 is fixed to the output shaft of the motor 706. The bevel spur gear 710 rotatably mounted on the mixing motor mounting bracket 702 is operatively coupled to the bevel gear 708 by the bevel gear teeth of the bevel spur gear 706 that mesh with the bevel gear teeth of the bevel gear 708. Thus, the rotation provided by the power of the bevel gear 708 about the rotating horizontal axis corresponding to the output shaft of the motor 706 is converted to the rotation of the bevel spur gear 710 about the vertical axis of rotation.

  The mixing motor assembly 700 is pivotally coupled to the underside of the blister plate 644 of the mounting plate 640 by a pivot screw 716 extending through the mixing motor bracket 702. A standoff 714 (comprising a threaded screw and a cylindrical sleeve disposed on a portion of the screw shaft) is attached to one end of the mounting bracket 702. A torsion spring 718 is coupled to the pivot screw 716 and biases the mixing motor assembly 700 inward relative to the side wall 474 (see FIG. 42), and the bevel spur gear 710 is coupled to the rotary mixer 192 of the composite cartridge 10 (see FIG. 8). The peripheral gear teeth 198 are engaged with each other.

  As shown in FIG. 48, the longitudinal spar 610 of the cam frame 606 includes a bevel block 616 extending inwardly from the longitudinal spar 610. As described above, the mixing motor assembly 700 is biased by the torsion spring 718 to pivot inwardly relative to the side wall 474 and the longitudinal spar 610. The bevel block 616 is positioned to engage the mixing motor assembly 700 when the cam frame 606 is in the forward position. Thus, when the cam frame 606 is in a retracted position that raises the heating and control assembly 500 to engage the composite cartridge 10 held within the cartridge carriage assembly 650, the mixing motor assembly 700 will engage the composite cartridge. Pivot inward under the force of torsion spring 718. As the cam frame 606 moves forward (to the left in FIG. 48) away from the composite cartridge held in the cartridge carriage assembly 650 and lowers the heating and control assembly 500, the bevel block 616 moves into the mixing motor. Mixing outward (toward the longitudinal spar 610) against the bias of the torsion spring 718 to contact the standoff 714 of the assembly 700 and disengage the bevel spur gear 710 from the rotary mixer 192 of the composite cartridge 10 Pivot the motor assembly. In one embodiment, before the actuator connector 618 of the cam frame 606 contacts the lever 658 of the cartridge latch 654 to lower the hook 656 and release the cartridge so that it can be ejected from the cartridge ejection assembly 670. The bevel block 616 contacts the standoff 714 to pivot the mixing motor assembly 700 out of engagement with the rotary mixer 192.

  Thus, when the cam frame 606 is in the forward position, the heating and control panel 500 is in a lower position where contact with the composite cartridge is removed and the magnet assemblies 552, 570 are moved away from the composite cartridge and into the retracted position. And the mixing motor assembly 700 pivots outward to disengage the composite cartridge, and the composite cartridge latch 654 pivots the hook 656 to disengage from the composite cartridge. Thus, the composite cartridge does not contact any of the components of the composite cartridge processing assembly 470, or otherwise does not engage, and the composite cartridge 10 can be ejected by the cartridge ejection assembly 670.

[Blister compression mechanism assembly (upper bay)]
Details of the blister compression mechanism assembly 750 are shown in FIG. 51, which is an exploded perspective view of the blister compression mechanism assembly 750. The assembly 750 includes a cam arm plate 752 and an array of cam arm motion compression mechanisms 754 operably mounted within the cam arm plate 752. The cam arm plate 752 is mounted on the top of the blister plate 644 of the mounting plate 640. The compression mechanism of the array 754 includes a compression mechanism configured to compress the rupturable fluid compartment or blister of the composite cartridge 10, a compression mechanism configured to compress the lance blister of the cartridge, and an active valve of the cartridge. A compression mechanism configured to push down and close the assembly. The various compression mechanisms of the array 754 are aligned with the blister holes 646 formed in the blister plate 644, and the compression mechanism of the array 754 is aligned with the blisters of the composite cartridge 10 disposed below the blister plate 644 in the processing bay 440. Access to the active valve is made possible.

  In various embodiments, LED PCB 466 is attached to cam arm plate 752.

  The blister compression mechanism assembly 750 further includes a cam follower plate 820 mounted on the cam arm plate 752 for linear motion relative to the cam arm plate. In various embodiments, one end of the cam follower plate 820 is secured to a linear guide rail 822 attached to the upper surface of the cam arm plate 752 by linear guide carriages 824a and 824b attached to the cam follower plate 820. The opposite end of the cam follower plate 820 is formed in the cam arm plate 752 by, for example, a suitable fastener within a recess 753 that includes a longitudinal slot 828 along one end thereof that receives the step end 830 of the cam follower plate 820. It is fixed against vertical movement by the mounted hold-down element 826 (or Z-axis constraint). Suitable materials for the construction of the holddown element include Delrin and brass. Therefore, the cam follower plate 820 is fixed in the Z or vertical or perpendicular direction with respect to the plane of the cam arm plate 752 in a predetermined space from the cam arm plate 752, and corresponds to the longitudinal direction of the linear guide rail 822. However, movement in a direction substantially parallel to the plane of the cam arm plate 752 is allowed, but movement in any direction across the linear guide rail 822 is restricted.

  The powered movement of the cam follower plate 820 relative to the cam arm plate 752 is effected by a cam follower plate motor 834 attached by a linear actuator 836 to a drive bracket 840 attached to the end of the cam follower plate 820. In various embodiments, the motor 834 further includes a rotary encoder 838 for providing precise control of the motor 834 and feedback from the motor 834. In various embodiments, the drive bracket 840 extends away from the attachment point to a cam follower plate 820 in a plane that generally corresponds to the plane of the cam follower plate, and the second portion extends from the cam follower plate. It has an “L” shape that extends downward in a direction substantially perpendicular to the plane. Linear actuator 836 is attached to drive bracket 840 at the lower end of the second, downwardly extending portion of drive bracket 840. This configuration of the drive bracket 840 limits the amount that the cam follower plate motor 834 extends over the cam follower plate 820, thereby maintaining the slim profile of the processing bay 440.

  In various embodiments, a sensor mechanism is provided to indicate when the cam follower plate 820 is in a specific, predetermined position with respect to the cam arm plate 752. In one embodiment, the sensor mechanism is mounted on the cam arm plate 752 and is contacted by the home switch contact surface 832 of the cam follower plate 820 when the cam follower plate 820 moves to the home position relative to the cam arm plate 752. A home switch 842 can be provided.

  In various embodiments, the cam arm plate 752 includes two light sensors 810, 812 that are arranged to spatially correspond to the locations of the inlet and outlet light ports 14, 16 (see FIG. 1), respectively. . The sensors 810, 812 are configured and arranged to detect fluid (eg, generate a signal) flowing through the inlet light detection chamber 154 and the outlet light detection chamber 158 of the sample preparation module 70 (FIG. 15). reference). The light sensors 810, 812 are connected to the LED PCB 466 and can be at least partially controlled by the LED PCB 466.

[Compression mechanism]
Details of the compression mechanism are shown in FIGS. 52, 53, and 54. FIG. 52 is a bottom partial plan view of the cam arm plate 752 showing the compression pads of the array 754 of compression mechanisms. FIG. 53 is a top perspective view of the compression mechanism of the array 754 isolated from the cam arm plate 752. 54 is a bottom perspective view of the compression mechanism of the array 754 isolated from the cam arm plate 752. FIG.

  The array 754 includes a plurality of fluid blister compression mechanisms, each configured to apply a compressive force on the associated deformable fluid blister when activated, thereby compressing the deformable blister. . In the illustrated embodiment, there are five fluid blister compression mechanisms 756a, 756b, 756c, 756d, and 756e corresponding to the deformable fluid chambers 34a, 36a, 38a, 40a, and 42a, respectively, of the composite cartridge. Exists.

  Array 754 further includes a plurality of lance blister compression mechanisms, each of which, when activated, applies a compressive force onto an associated lance blister associated with one of the deformable fluid blisters, thereby compressing the lance blister. And a fluid seal in the lance blister. In the illustrated embodiment, there are five lance blister compression mechanisms 760a, 760b, 760c, 760d, and 760e corresponding to the lance blisters 34b, 36b, 38b, 40b, and 42b, respectively, of the composite cartridge.

  The array 754 has substantially the same configuration as the lance blister compression mechanisms 760a-e and further includes a compression mechanism 758 corresponding to the blister 44 of the composite cartridge.

  Array 754 includes two valve actuator compression mechanisms 762a, 762b associated with sample valve assembly 204 and waste valve assembly 219, respectively (see FIG. 15). Each of the valve actuator compression mechanisms 762a, 762b, when activated, applies a compressive force to the valve actuator tabs 20, 18, respectively (see FIG. 1), thereby activating and closing the active valves 219 and 204. Configured as follows.

  Details of the configuration of each of the various compression mechanisms are shown in FIGS. 53 and 54, along with FIGS. 55A, 55B, and 55C. FIG. 55A is an exploded perspective view of a single fluid blister compression mechanism. FIG. 55B is an exploded perspective view of a single lance blister compression mechanism. FIG. 55C is an exploded perspective view of the valve actuator compression mechanism.

  The blister compression mechanism assembly uses the principles and concepts described in US Patent Application No. 14 / 206,817, entitled “Apparatus and Methods for manipulating deformable fluid vessels”, the contents of which are hereby incorporated by reference. . In particular, the blister compression mechanism assembly is a fluid blister, lance blister, and valve assembly without requiring air, electromechanical or other components at greater distances above and / or below the composite cartridge 10. Is configured and arranged to convert the horizontal movement of the cam follower pate 820 into a vertical or partially vertical compression mechanism movement, and maintains the slim profile of the processing bay 440.

  Referring to FIG. 55A, each fluid blister compression mechanism, such as fluid blister compression mechanism 756a, includes a cam arm 764 having a cam surface 766 formed along the upper end. Cam arm 764 is mounted within cam arm plate 752 for pivoting motion about arm pivot pin 768 extending through a hole formed in one end of cam arm 764. The cam arm 764 is disposed in a slot 765 formed in the cam arm plate 752, and the arm pivot pin 768 is mounted in the cam arm plate 752 laterally in the slot (see FIG. 52). The compression pad 772 is pivotally mounted at the opposite end of the cam arm 764 for pivoting motion about a pad pivot pin 774 extending through a hole formed at the opposite end of the cam arm 764. In various embodiments, the compression pad 772 is disposed in a blind recess 773 formed in the lower surface of the cam arm plate 752 that has a shape that substantially matches the shape of the compression pad 772 (see FIG. 52).

  The fluid blister compression mechanism 756a has an arm pivot pin 768 between a retracted position where no pressure is applied to the fluid blister with which the compression mechanism is associated and an extended or deployed position where the compression mechanism applies a compressive force to the fluid blister. It is configured to pivot about a cam arm plate 752. Torsion spring 770 biases compression mechanism 756a to the retracted position. In the retracted position, the cam arm 764 is substantially disposed in a corresponding slot 765 formed in the cam arm plate 752, and the compression pad 772 is disposed in a pad recess 773 formed in the cam arm plate 752. The blister contact surface of the compression pad 772 is substantially flushed with the surface of the cam arm plate 752. In the extended position, the cam arm 756 rotates about the cam arm pivot pin 768 and the compression pad 772 underneath the cam arm plate 752 to compress and rupture the reagent blister located under the compression pad 772. Stretched.

  The cam surface 766, in various embodiments, can include a convex bulge or other feature that extends over the top surface of the cam arm plate 752 (see FIG. 51, compression mechanism extending over the cam arm plate 752). The cam features of the cam arms of the array 754 of FIG. When the cam surface 766 is engaged by a cam follower element that moves relative to the cam arm 764 above the cam surface 766, the cam arm 764 moves from the retracted position to the extended position when the cam follower moves on the convex bulge of the cam surface 766. It is made to pivot. When the cam follower element moves away from the cam surface 766, the cam arm 764 returns to the retracted position under the force of the torsion spring 770.

  The cam arm 764 is preferably made of a material that has sufficient strength to withstand the force applied by the cam follower element that pushes the cam arm 764 against the rupturable fluid blister and that has adequate machinability. Suitable materials include steel for applications where the cam follower element comprises a roller that rolls over the cam surface 766. For applications where the cam follower element comprises a sliding (ie non-rolling) element that slides on the cam surface 766, suitable materials are low friction, low wear materials such as nylon or oil-containing bronze. Contains lubricating oil material.

  In various embodiments, the structure and operation of the other fluid blister compression mechanisms 756b, 756c, 756d, and 756e are substantially the same as those of the fluid blister compression mechanism 756a, but a compression pad (eg, a compression pad) 772) may vary from one fluid blister compression mechanism to another depending on the size and shape of the fluid blister to be compressed by the compression mechanism.

  Referring to FIG. 55B, each lance blister compression mechanism, such as lance blister compression mechanism 760a, includes a cam arm 780 having a cam surface 782 formed along the upper end. Cam arm 780 is mounted within cam arm plate 752 for pivoting motion about arm pivot pin 784 extending through a hole formed in one end of cam arm 780. The cam arm 780 is disposed in a slot 781 formed in the cam arm plate 752, and the arm pivot pin 784 is mounted in the cam arm plate 752 across the slot (see FIG. 52). The compression pad 788 is formed and disposed at the opposite end of the cam arm 780. In various embodiments, the compression pad 788 is disposed in a blind recess 789 formed in the lower surface of the cam arm plate 752 that is shaped to substantially match the shape of the compression pad 788 (see FIG. 52).

  The lance blister compression mechanism 760a has an arm pivot pin 784 between a retracted position where no pressure is applied to the lance blister with which the compression mechanism is associated and an extended or deployed position where the compression mechanism applies a compressive force to the lance blister. Around it is configured to pivot relative to the cam arm plate 752. The torsion spring 786 biases the compression mechanism 760a to the retracted position. In the retracted position, the cam arm 780 is substantially disposed in a corresponding slot 781 formed in the cam arm plate 752, and the compression pad 788 is disposed in a pad recess 789 formed in the cam arm plate 752. The blister contact surface of the compression pad 788 is substantially cleaned with the surface of the cam arm plate 752. In the extended position, the cam arm 780 is rotated about the cam arm pivot pin 784 so that the compression pad 788 compresses and ruptures the lance blister disposed under the compression pad 788. Stretched down.

  The cam surface 782 may, in various embodiments, include a convex bulge or other feature that extends over the top surface of the cam arm plate 752 (see FIG. 51, of a compression mechanism that extends over the cam arm plate 752). The cam features of the cam arms of array 754 are shown). When the cam surface 782 is engaged by a cam follower element that moves relative to the cam arm 780 on the cam surface 782, the cam arm 780 moves from the retracted position to the extended position when the cam follower moves on the convex bulge of the cam surface 782. It is made to pivot. When the cam follower element moves away from the cam surface 782, the cam arm 780 returns to the retracted position under the force of the torsion spring 786.

  The cam arm 780 is preferably made of a material that has sufficient strength to withstand the force applied by the cam follower element that pushes the cam arm 780 against the rupturable lance blister and has adequate machinability. Suitable materials include steel for applications in which the cam follower element comprises a roller that rolls over the cam surface 782. For applications in which the cam follower element includes a sliding (ie, non-rolling) element that slides on the cam surface 782, suitable materials include low friction, low wear materials such as nylon or oil bronzes. Contains lubricating oil material.

  In various embodiments, the structure and operation of lance blister compression mechanisms 760b, 760c, 760d, and 760e and compression mechanism 758 are substantially the same as those of lance blister compression mechanism 760a.

  Referring to FIG. 55C, each valve actuator compression mechanism, such as valve actuator compression mechanism 762a, includes a cam arm 790 having a cam surface 792 formed along its upper end. The cam arm 790 is mounted in a cam arm plate 752 for pivoting movement about a pivot pin 794 that extends through a hole formed in one end of the cam arm 790. The cam arm 790 is disposed in a slot 791 formed in the cam arm plate 752, and the arm pivot pin 794 is mounted in the cam arm plate 752 crossing the slot (see FIG. 52). Contact pads 798 are formed or disposed at opposing ends of the cam arm 790. In various embodiments, the contact pad 798 is disposed within a blind recess 799 formed in the lower surface of the cam arm plate 752 having a shape that substantially matches the shape of the contact pad 798 (see FIG. 52).

  In various embodiments, the contact pad 798 can further include a contact pin or contact point 800 that protrudes from the contact pad 798. To prevent the compression mechanism from slipping out of the valve actuator tab, when the valve actuator compression mechanism is pressed against the tab, the contact point is a small excavator formed on the upper surface of the valve actuator tab 18 or 20. Or it is configured to mesh with the indentation. Also, in various embodiments, a portion of the contact pad 798 and the contact pin 800 can be offset from the cam arm 690 to accommodate spatial and directional constraints within the array 754 of compression mechanisms.

  The valve actuator compression mechanism 762a includes a retracted position where the compression mechanism does not apply pressure to the associated valve actuator tab and the active valve assembly, and an extended or deployed position where the compression mechanism applies a compressive force to the actuator tab and valve assembly. It is configured to pivot relative to the cam arm plate 752 about an arm pivot pin 794 therebetween. The torsion spring 796 biases the compression mechanism 762a to the retracted position. In the retracted position, the cam arm 790 is substantially disposed in a corresponding slot 791 formed in the cam arm plate 752 and the contact pad 798 is disposed in a pad recess 799 formed in the cam arm plate 752. The contact surface of the contact pad 798 is substantially cleaned with the surface of the cam arm plate 752. In the extended position, the cam arm 790 is rotated about the cam arm pivot pin 794 so that the contact pad 798 extends below the cam arm plate 752 and bends the valve actuator tab downwardly, below the valve actuator tab. Close the associated valve assembly located in

  The cam surface 792 may include a convex bulge or other feature that, in various embodiments, extends over the top surface of the cam arm plate 752 (see FIG. 51, extends over the cam arm plate 752). The cam features of the cam arms of the array 754 of compression mechanisms are shown). When the cam surface 792 is engaged by a cam follower element that moves relative to the cam arm 790 on the cam surface 792, when the cam follower moves on the convex bulge of the cam surface 792, the cam arm 790 moves from the retracted position to the extended position. Pivoted. When the cam follower element moves away from the cam surface 982, the cam arm 790 is returned to the retracted position under the force of the torsion spring 796.

  The cam arm 790 is preferably made of a material that has sufficient strength to withstand the force applied by the cam follower element that pushes the cam arm 790 against the valve assembly and that has suitable machinability. Suitable materials include steel for applications in which the cam follower element comprises a roller that rolls over the cam surface 792. For applications in which the cam follower element includes a sliding (ie, non-rolling) element that slides on the cam surface 792, suitable materials include low friction, low wear materials such as nylon, or oil-containing bronze. Contains lubricating oil material.

  In various embodiments, the configuration and operation of the other valve actuator compression mechanism 762b is substantially the same as that of the valve actuator compression mechanism 762a.

  Details of the cam follower plate 820 are shown in FIGS. 56 is a lower plan view of the cam follower plate 820, and FIG. 57 is a lower perspective view of the cam follower plate 820.

  Cam follower plate 820 includes a plurality of generally parallel, elongated cam grooves 850, 852, 854, 856, 858, and 860. Each of the grooves 850-860 of the cam follower plate 820 receives a portion of one or more cam arms 764, 780, 790 of the compression mechanism of the array 754. Further, each groove 850-860 includes one or more cam follower elements, for example, in the form of ribs or rollers that are formed or arranged at discrete locations along the corresponding groove.

  As described above, the cam follower plate 820 is configured for linear operation with respect to the cam arm plate 752 in a plane parallel to the cam arm plate 752. When the cam follower plate 820 moves with respect to the cam arm plate 752, the cam follower element in the cam groove moves to the cam surface of the cam arm of the compression mechanism (for example, the cam surfaces 766, 782, or 792 of the cam arms 764, 780, or 790, respectively). ), The cam arm is pushed downward and pivots about a respective arm pivot pin (eg, pivot pin 768, 784, or 794), causing the compression mechanism to blister (eg, compressed fluid blister or lance). Blister) or push an active valve assembly placed under the compression mechanism.

  During movement of the cam follower plate 820 relative to the cam arm plate 852, the relative position of the compression mechanism of the array of compression mechanisms 754 and the cam follower ribs formed in the grooves 850, 852, 854, 856, 858, and 860 is determined by the compression mechanism. Specifies the sequence in which is activated.

[Software and hardware]
Generally and specifically as described above, aspects of this disclosure are implemented by control and computing hardware components, user-generated software, data input components, and data output components. A hardware component receives one or more input values and executes one or more algorithms (eg, software) stored on a non-transitory machine-readable medium that provides instructions for operations or otherwise actions on the input values Computing and / or control modules (eg, system controller (s)), such as microprocessors and computers, configured to enable computation and / or control steps and to output one or more output values )including. Such output can be displayed to the user or otherwise shown to provide the user with information such as, for example, information about the status of the device or processes performed thereby, or , Such outputs can include inputs to other processes and / or control algorithms. The data input component comprises elements into which data is input for use by control and computing hardware components. Such data may comprise position sensors, motor encoders, as well as manual input elements such as graphic user interfaces, keyboards, touch screens, microphones, switches, manual scanners, voice sensitive inputs. The data output component can comprise a hard drive or other storage medium, graphic user interface, monitor, printer, indicator light, or audio signal element (eg, buzzer, horn, bell, etc.).

  The software includes instructions stored on a non-transitory computer readable medium that, when executed by the control and computing hardware, cause the control and computing hardware to perform one or more automatic or semi-automated processes. .

[Sample preparation process]
An exemplary sample preparation process that can be performed in the sample preparation module 70 is described and illustrated in FIGS. Sample preparation processes other than those described herein-for example, permutation of steps from those described herein, omission of certain steps described herein, and / or addition of certain steps- Those skilled in the art will recognize that can be performed by the sample preparation module or a modified version of the sample preparation module.

  In the first step, illustrated in FIG. 16, a fluid sample sample is sent to the sample well 78. In general, the composite cartridge 10 is designed for processing liquid or solid samples. Liquid samples can include epithelial samples such as blood, serum, plasma, urine, saliva, cerebrospinal fluid, lymph, sweat, semen, or buccal, nasopharyngeal, anal or vaginal swabs, and cells Lysis buffer is added to resuspend. Solid samples such as feces or tissue samples (eg, tumor biopsy) generally need to be resuspended and diluted in a buffer such as, for example, Cary Blair transport medium. The sample well 78 can then be closed using the sample cap 84 (see FIG. 6), and the composite cartridge 10 can then be transferred to the processing device (eg, in the processing bay 440 of the processing module 410 of the device 400). Arranged).

  As illustrated in FIG. 17, in a first step performed within the apparatus, the lance blister 34b associated with the deformable compartment 34a pushes the bead or other opening device through the closure seal (ie, the bead or Compressed by an external actuator (e.g., compression mechanism 760a) to break the seal with other devices, and then the deformable compartment 34a is there toward the first inlet port 126 formed in the substrate 72. In order to apply force to the contained processing fluid, it is compressed by an external actuator (eg, compression mechanism 756a). In one embodiment, the processing fluid contained in the deformable compartment 34a is a lysis buffer. Fluid is directed from the inlet port 136 to the sample well 78 by the first fluid channel 150, where the fluid enters the sample well 78 via the inlet snorkel 180. In addition, an external pump (eg, pump 458) connected to the sample preparation module 70 at the pump port 104 generates pressure that is applied to the contents of the sample well 78 via the pressure conduit 106.

  The pressure generated by compressing the deformable compartment 34a and the pressure applied in the pressure conduit 106 pushes the fluid contents--including the fluid sample and the contents of the deformable compartment 34a--from the sample well 78. Two fluid channels 152 lead to the lysis chamber inlet 122. The fluid continues to flow through the lysis chamber and exits the outlet 124 where it is directed into the mixing well 90 by the third fluid channel 156 and a portion of the fifth fluid channel 162. As the fluid stream first enters or exits the lysis chamber 120 and passes through the entrance light detection chamber 154 or the exit light sensor chamber 158, the light detection (s) mounted on the light detector (eg, LED PCB 466). ) Through the associated optical port 14 or 16 (see FIG. 1) formed in the top beam 12. A signal (e.g., an air flow interface) from the photodetector indicating fluid flow through the inlet or outlet light sensing chambers 154 or 158 activates the lysis chamber mixer motor 128 to cause the fluid flowing through the lysis chamber 120 to flow. The lysis beads contained in the lysis chamber 120 are interrupted. End of fluid flow through the inlet or outlet light sensing chamber 154 or 158-Thus, the motor 128 continues to operate until a signal from the photodetector indicating the end of flow through the lysis chamber 120 stops the motor 128 from operating. .

  As the fluid mixture flows into the mixing compartment 90, the passive valve port 108 remains open, preventing the pressure in the mixing well 90 from rising to a level that would open the passive valve assembly 220. Thus, at the end of the step illustrated in FIG. 17, the mixing well 90 is the contents of the deformable compartment 34a that is physically lysed by the fluid sample and lysis beads contained in the lysis mixer and lysis chamber 120. In admixture (eg lysis buffer).

  Referring now to FIG. 18, after the steps shown in FIG. 17, the air pump that applies pressure to the pressure port 104 is turned off, for example, after a predetermined period of operation, and the third deformable compartment 44. Are compressed by an external actuator (eg, compression mechanism 758) to allow the contents of the deformable compartment 44 to go into the third inlet port 140. In one embodiment, the contents of the deformable compartment 44 comprise magnetic object capture beads.

  The lance blister 36b associated with the deformable compartment 36a then compresses the bead or other opening device through the closure seal (ie, breaks the seal with the bead or other device) (external actuator ( For example, compression mechanism 760e) compresses the deformable compartment 36a from an external actuator (eg, compression mechanism) to push processing fluid contained therein to a second inlet port 138 formed in substrate 72. 756e). The processing fluid then flows to the mixing well 90 through the fourth fluid channel 160 and the fifth fluid channel 162. The contents of the deformable compartment 36a can include a binding buffer to facilitate binding of the target capture beads to the target analyte (s). Under pressure generated by compression of the deformable compartment 36a, the fluid flowing through the third inlet port 140 causes the fluid contents of the deformable compartment 36a and the contents of the deformable compartment 44 to be The five fluid channels 162 are transported to the mixing well 90.

  As described above, in another embodiment, the magnetic beads can be provided in the form of lyophilized pellets contained within the mixing well 90 and the deformable compartment 44 and associated external actuators (e.g., , Rupturing the compression mechanism 758) and the deformable compartment 44 can be omitted.

  After the steps illustrated in FIG. 18 are completed, the rotor mixer 192 in the mixing well 90 can be activated (eg, by the mixing motor assembly 700) to agitate the contents of the mixing well 90. In various embodiments, lyophilized or other dried reagent forms can be pre-placed in the mixing well 90 and lysed or reconstituted by the fluid transported to the mixing well 90. The rotary mixer 192 facilitates the dissolution or reconstitution of the dry reagent and helps to mix all materials contained in the mixing well to form a uniform fluid mixture.

  Referring to FIG. 19, the next step involves rupturing the lance blister 38b associated with the deformable compartment 38a (eg, with the compression mechanism 760b), thereby opening the compartment into the fourth inlet port 142. . The deformable compartment 38a is then ruptured (eg, by a compression mechanism 756b), directing its fluid content to the fourth inlet port 142, through the sixth fluid channel 164, and the first outlet port. Go to 182 and the fluid exits the sample preparation module 70. The first outlet port 182 communicates with the inlet port 252 of the reaction module 240 as described above. The fluid contained in the deformable compartment 38a can include, for example, an insoluble fluid such as oil, which is a reaction module 240 between the top plate 241 and the fluid treatment panel 354, as shown in FIG. Used to fill the reaction space 295 inside.

  Referring now to FIG. 20, the lance blister 40b associated with the deformable compartment 40a is ruptured by an external actuator (eg, compression mechanism 760c) to open the compartment to the fifth inlet port 144, and then The deformable compartment 40a is ruptured by an external actuator (eg, compression mechanism 756c) to allow its fluid contents to go to the fifth inlet port 144. The fluid content flows from the fifth inlet port 144 to the second outlet 188 via the seventh channel fluid 166. In one embodiment, the fluid content of the deformable compartment 40a is shown in FIG. 31, and as described above, from the second exit port 188 via the inlet 278, the rehydration buffer of the reaction module 240. Contains rehydration or elution buffer flowing into compartment 276. The same buffer solution contained in the deformable compartment 40a is used for both rehydration of dried or lyophilized reagents or other substances and elution of nucleic acids or other analytes of interest from the bound substrate. Can be done.

  Referring now to FIG. 21, in the next step, the active valve assembly 204 is closed by an external actuator (eg, valve actuator 762b) that depresses the valve. An air pump coupled to pump port 104 includes pressure conduit 106, a portion of first fluid channel 150, a second fluid channel 152, a third fluid channel 156, and a portion of fifth fluid channel 162. Is activated to apply pressure to the mixing well 90. At the same time, the passive valve port 108 is closed to allow the pressure in the mixing well 90 to rise, which will actuate the passive valve assembly 220, thereby opening the passive valve 220 and passing through the capture compartment 100. Thus, the fluid content of the mixing well 90 can flow through the channels 92 and 172. Fluid flowing through the capture compartment 100 flows through the thirteenth fluid channel 178, but is stopped from flowing into the fourteenth fluid channel 180 by the closed active valve assembly 204. The active valve assembly 219 remains open, allowing fluid in the thirteenth fluid channel 178 to flow into the tenth fluid channel 172 and the waste chamber 102. While fluid flows through the capture compartment 100, the contents are exposed to a magnetic force, for example, by placement of an external magnet in the vicinity of the capture compartment 100 (eg, by deploying the sample preparation magnetic assembly 570). The magnetic force retains the magnetic target capture beads and the target analyte (s) (eg, nucleic acid (s)) bound thereto in the capture compartment 100 while the rest of the contents of the mixing well 90 remain. Through the capture compartment 100 to the waste chamber 102.

  Referring now to FIG. 22, in the next step, the valve assembly 204 remains closed, the valve assembly 219 remains open, and the lance blister 42b associated with the deformable compartment 42a (eg, compression mechanism) Rupture (by 760d), thereby opening the compartment to the sixth inlet port 146. The deformable compartment 42a is then partially ruptured (eg, by the compression mechanism 756d) and sends a portion of the contents (eg, about 50%) to the sixth inlet port 146. In one embodiment, the fluid content of the deformable compartment 42 a includes a wash buffer that flows from the sixth inlet port 146 to the capture compartment 100 via the twelfth fluid channel 176. The wash fluid flows over the capture beads that are secured within the capture compartment 100 (eg, by a magnet) and through channels 178, 172, and 174 to the wash chamber 102, thereby unbound material and Transport other debris to the waste chamber 102.

  Referring now to FIG. 23, in the next step, the waste valve assembly 219 is closed by an external actuator (eg, valve actuator 762a) and the sample valve assembly 204 is opened by removing the external actuator. The remainder of the deformable compartment 42a is then ruptured by an external actuator (eg, compression mechanism 756d), thereby capturing the remainder of the fluid (eg, wash buffer) through the twelfth fluid channel 176. Go into the compartment 100. The magnetic force is removed from the capture compartment 100 (eg, by retracting the sample preparation magnet assembly 570), the magnetic beads in the capture compartment 100 are released, pass through the capture compartment 100, through the thirteenth fluid channel 178, It can be carried by the fluid flowing through the sample valve assembly 204 and the fourteenth fluid channel 180 to the third outlet 190. The fluid flowing from the third outlet 190, now containing at least partially purified analyte of interest carried by the magnetic beads, is shown in FIG. 31 and, as described above, via the inlet 268, of the reaction module 240. Sent into the sample compartment 266.

  56 and 57, each of the cam follower ribs formed in the cam grooves 850 to 860 of the cam follower plate 820 is indicated by unique parenthesized numbers (1) to (14). When the cam follower plate 820 is moved relative to the cam arm plate 852 in the direction “A”, the cam follower ribs formed in the various cam grooves 850 to 860 are arranged in a predetermined sequence in the compression mechanism of the actuator array 754. , Open various reagent chambers, send their contents, and activate various activity values in a specific sequence. The bracketed numbers assigned to the cam follower ribs of FIGS. 56 and 57 indicate that each rib is in contact with the associated cam arm of the compression mechanism of the array 754, as described above and shown in FIGS. FIG. 6 shows a sequence for operating the compression mechanism in a sequence corresponding to the sample preparation process performed in module 70. FIG. Table 1 below shows between each cam follower rib of the cam follower plate 820, process steps, corresponding compression mechanism, and compressible burstable chamber or active valve of the composite cartridge 10 for the process shown in FIGS. The correspondence of is shown.

* Step (2) may optionally be omitted if magnetic beads are provided directly in the mixing well 90, for example by lyophilized pellets.

[Sample reaction process]
Sample material sent from the sample processing module 70 to the sample compartment 266 of the reaction module 268 undergoes a reaction process in the reaction module 240. In one exemplary embodiment, the reaction process includes PCR amplification and analyte detection.

  An exemplary process will be described with reference to flowchart 900 of FIG. Although the various elements (steps) of the flowchart 900 of FIG. 60 are shown as sequential steps having a defined order, the illustrated process 900 is exemplary and is intended to be limiting. It should be understood that there is no. Many of the various elements (steps) of process 900 can be performed in a different order than that illustrated and described herein, and at the same time or substantially other elements (steps). Those skilled in the art will recognize that they can be performed simultaneously or can be omitted altogether. Thus, the order of the elements (steps) shown in FIG. 60 is (unless specifically specified or otherwise suggested by the context of the specification) unless a particular order of two or more elements (steps) is specified. For example, a mixture should be first formed before the mixture can be cultured or otherwise manipulated) and should not be considered limiting.

  In step S1, an aliquot (eg, 15 μl) of elution / reconstitution buffer is added to the rehydration buffer area 372 (FIG. 59) (and the rehydration buffer compartment 376 of the top plate 241 (FIGS. 26, 27). )) To the electrowetting path defining the exonuclease zone 384 (FIG. 59) by electrowetting droplet manipulation.

  As described above, in an embodiment of the present invention, the region of the reaction module 240 between the top plate 241 and the fluid treatment panel 354 can be filled with a treatment fluid such as an insoluble fluid such as oil. The drops are manipulated via oil.

  In step S2, an amount of sample mixture (including magnetic beads with DNA material bound thereto and wash solution from sample preparation module 70) is added to sample bead area 368 (FIG. 59) (and top plate 241 ( While held by the electrowetting operation in the sample compartment 266) of FIGS. 26, 27), the magnetic beads are held in the sample bead area 368 by a magnet focused at location 369 (referred to as the bead collection region). Withdrawn from the resulting aqueous solution. When the cartridge magnet assembly 552 is in the deployed position, the bead collection region 369 corresponds to the position of the focusing magnet 558 of the cartridge magnet assembly 552 (see FIG. 49B) adjacent to the fluid treatment panel 554 of the composite cartridge 10. During the process of collecting the magnetic beads in the bead collection area 369, the aqueous solution selects a different electrowetting pad to move the aqueous droplet containing the magnetic beads closer to the magnetic force in the bead collection area 369. Can be moved across the sample bead area 368 by dynamic operation.

  In step S3, sample waste (ie, wash buffer and other materials from which magnetic beads are removed in step S2) is retained in the sample bead area 368 (and sample compartment 266) by electrowetting droplet manipulation. Thereby separating the magnetic beads and target analyte material bound thereto from other constituents of the sample bead mixture dispensed from the sample preparation module 70 to the sample bead area 368.

  In step S4, an amount of reconstitution buffer sent from the rehydration buffer area 372 in step S1 is added to the PCR reagent area 376 (FIG. 59) (and top plate) by electrowetting droplet manipulation. 241 buffer compartment 296 (FIGS. 26, 27)). The resuspension of the dry PCR reagent contained within the PCR reagent area 376 occurs due to vibrational movement of the droplets between the electrowetting pads within the PCR reagent area 376.

  In step S5, an amount of reconstitution buffer sent from the rehydration buffer area 372 and not transported to the PCR reagent area 376 is retained by the magnetic force in the bead collection area 369 for the final bead wash. Is transported by electrowetting droplet manipulation on magnetic beads. After the final bead wash, the reconstitution buffer is moved by electrowetting droplet manipulation to the end of the central pathway corresponding to exonuclease zone 384, where it is bead collected by electrowetting droplet manipulation. It is held away from the magnetic beads held in region 369.

  In step S6, the reconstituted PCR buffer in PCR reagent area 376 is dispensed to the respective primer cocktail positions of thermal cycling tracks 364a, 364b, 364c, and 364d by electrowetting droplet manipulation. One primer cocktail position 366a at the proximal end of the thermal cycle track 364d is labeled in FIG. Each of the other thermal cycling tracks 364a, 364b, and 364c has a similar primer cocktail location. The reconstituted PCR reagent combination with the dried primer cocktail at the primer cocktail position (eg, position 366) reconstitutes the primer cocktail at that position. In this configuration, the reaction module 240 is configured to perform one PCR reaction on each of the thermal cycle tracks 364a, 364b, 364c, and 364d.

  In another embodiment, a primer cocktail can also be provided at the far end of each thermal cycling track 364a, 364b, 364c, and 364d. One primer cocktail position 366b at the far end of the thermal cycle track 364d is labeled in FIG. Each of the other thermal cycling tracks 364a, 364b, and 364c can have similar primer cocktail locations. In such a configuration, the reaction module 240 is configured to perform two PCR reactions on each of the thermal cycle tracks 364a, 364b, 364c, and 364d.

  In step S7, the magnetic force is removed from the bead collection region 369 (eg, by moving the cartridge magnet assembly 552 to the retracted position). The reconstitution / elution buffer is moved from the central path 384 to the bead collection area 369 by electrowetting droplet manipulation, and the magnetic bead and reconstitution / elution buffer mixture from the rehydration buffer area 372 is The electrowetting droplet operation reciprocates along path 384 to elute the DNA material (or other analyte of interest) from the magnetic beads.

  After a sufficient elution period, in step S8, the cartridge magnet assembly 552 is again deployed to apply a magnetic force (via the converging magnet 558) to the bead collection region 369 to remove the magnetic beads from which the DNA material has been eluted. The drawn, held (fixed) and eluted DNA material is transferred to the PCR staging area at the proximal end of each of the thermal cycling tracks 364a, 364b, 364c, and 364d by electrowetting droplet manipulation. In the embodiment and orientation shown in FIG. 59, the PCR staging area is at the left end of the thermal cycling tracks 364a, 364b, 364c, and 364d.

  In step S9, the PCR droplet—including the eluted DNA material, the reconstituted PCR reagent, and the reconstituted PCR primer—is converted into each PCR of the thermal cycle tracks 364a, 364b, 364c, and 364d by electrowetting droplet operation. It is formed in the staging area. Each PCR droplet is moved to a corresponding one of the thermal cycle tracks 364a, 364b, 364c, and 364d, and the PCR process is performed in a PCR (thermal cycle) region 382a (for annealing and extension, eg, about 60 ° C. ) And 382b (for denaturation, eg, about 95 ° C.) or 382c (about 60 ° C. for annealing and extension) and 382b (for denaturation, eg, about 95 ° C.) ) By reciprocating the droplet between the two. In other embodiments, two PCR droplets are transported to each thermal cycle track 364a, 364b, 364c, and 364d, and one droplet is shuttled between the heater regions 382c and 382b and the other liquid Drops are shuttled between heater regions 382a and 382b. The PCR process can continue for about 40 minutes or less.

  In step S10, an amount of elution / reconstitution buffer is sent from the rehydration buffer area 372 by electrowetting droplet manipulation, and resuspension of the dry exonuclease reagent by electrowetting droplet manipulation. For transport to the exonuclease reagent zone 374 (FIG. 59) (and the second buffer compartment 300 of the top plate 241 (FIGS. 26, 27)). The resuspension of the dry exonuclease reagent contained within the exonuclease reagent zone 374 occurs due to vibrational movement of the droplets between the electrowetting pads within the exonuclease reagent zone 374. The reconstituted exonuclease reagent is then transported from the exonuclease reagent zone 374 to the PCR staging area of thermal cycling tracks 364a, 364b, 364c, and 364d by electrowetting droplet manipulation.

  In step S11, following the PCR (step 9), each droplet that has undergone the PCR process is mixed with an amount of exonuclease drug that is resuspended in step S10, and an electrowetting droplet operation results in an exonuclease zone. Transported to 384 and held in a separate location within exonuclease zone 384. In various embodiments, an amount of elution / reconstitution buffer from the buffer zone 372 is added to each PCR droplet by electrowetting droplet manipulation to provide a suitable amount of total volume for each droplet. Up to.

  In step S12, the droplet mixture formed in step S11 includes the PCR product and the reconstituted exonuclease reagent, and then within the exonuclease region 380 and exonuclease zone 384 at a predetermined temperature and for a predetermined period of time. Incubate.

  In step S13, the detection reagent in the hybridization zone 370 (FIG. 59) (and the detection buffer compartment 280 of the top plate 241 (FIGS. 26, 27)) has an amount from the rehydration buffer zone 372. Reconstituted with rehydration buffer. In one embodiment, an amount of rehydration buffer from rehydration buffer area 372 is detected via connection path 274 (FIGS. 26, 27) via electrowetting droplet manipulation. It is moved between the buffer compartment 280 and the rehydration buffer compartment 276.

  In step S14, an amount (eg, 25 μl) of reconstituted detection reagent from the hybridization zone 370 is mixed with each of the PCR droplets by electrowetting droplet manipulation. Each PCR droplet is then mixed with a signal probe cocktail stored at locations 362a, 362b, 362c, and 362d of the fluid treatment panel 354. In order to enable mixing of the PCR droplets and signal probe cocktail and to resuspend the signal probe cocktail, each droplet is transferred to the detection mixing zone 385a, 385b, 385c, and 385d by electrowetting droplet manipulation. It can be transported around or in one.

  In step S15, the droplets are transported to the electronic sensor arrays 363a, 363b, 363c, and 363d by an electrowetting operation, where they undergo further culturing within the detection region 378 and are of various interests. Such analytes are detected by electronic sensing techniques as described above and / or described in published documents incorporated by reference.

Exemplary Embodiments The following embodiments are included in the foregoing disclosure.

Embodiment 1. FIG.
A substrate,
A sample well formed in the substrate and configured to receive a volume of fluid sample;
A closure configured to be selectively placed on the sample well;
In a supported and undeformed state on the substrate, the fluid is retained therein and is configured to rupture and discharge at least a portion of the fluid from the fluid chamber when an external compressive force is applied. A deformable fluid chamber through which fluid flows through the sample well and the channel through a channel formed in the substrate;
A first formed in the substrate and extending through the sample well and fluid through the channel formed in the substrate, extending from the first peripheral wall defining the well, the first floor, and the channel leading to the mixing well to the sample well. A fluid inlet snorkel that extends over one of the peripheral walls of the first and stops under the upper end of the first peripheral wall;
A drive mixing device disposed in the mixing well and configured and arranged to mix the contents of the mixing well; and
A fluid sample processing cartridge comprising:

Embodiment 2. FIG. The fluid sample processing cartridge of embodiment 1, wherein the fluid inlet snorkel extends on an outer surface of the first peripheral wall and stops at an opening formed in the first peripheral wall.

Embodiment 3. FIG. The sample well comprises a second peripheral wall and a second floor that define the well, and a fluid inlet snorkel that extends across one surface of the second peripheral wall and stops under the upper end of the second peripheral wall. The fluid sample processing cartridge according to the first or second embodiment.

Embodiment 4 FIG. Any of Embodiments 1-3, wherein the mixing well further comprises an exit port comprising one or more openings formed in the second floor, the floor being tapered downward toward the exit port. The fluid sample processing cartridge according to claim 1.

Embodiment 5. FIG. The drive mixing device meshes with a first impeller that is rotatably arranged in the mixing well so that it can be driven by the meshing gear of the device into which the fluid sample processing cartridge is inserted, and when the meshing gear meshes, A fluid sample processing cartridge according to any one of embodiments 1-4, comprising: a gear configured to rotate the first impeller.

Embodiment 6. FIG.
A lysis chamber comprising a plurality of lysis beads, formed on the substrate and disposed along a channel connecting the mixing well and the sample well, wherein fluid flowing from the sample well to the mixing well flows through the lysis chamber. ,
Embodiments 1-5 further comprising a bead mixer disposed at least partially within the lysis chamber and configured and arranged to agitate the lysis beads and the fluid flowing through the lysis chamber. A fluid sample processing cartridge according to claim 1.

Embodiment 7. FIG.
A first optical interface comprising an enlarged portion of a channel connecting the lysis chamber to the sample well;
A second optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the mixing well;
The fluid sample processing cartridge of embodiment 6, further comprising:

Embodiment 8. FIG. The bead mixer
A motor mounted on the board;
A second impeller disposed in the dissolution chamber and mounted on the output shaft of the motor;
A fluid sample processing cartridge according to embodiment 6 or 7, comprising:

Embodiment 9. FIG. Embodiments 6-8, wherein the lysis chamber includes a fluid inlet and a fluid outlet, further comprising a mesh filter disposed on each of the fluid inlet and the fluid outlet and configured to retain lysis beads in the lysis chamber. The fluid sample processing cartridge according to claim 1.

Embodiment 10 FIG.
A pressure port formed in the substrate and configured to couple the substrate to an external fluid pressure source;
A channel formed in the substrate and connecting the pressure port to the sample well;
The fluid sample processing cartridge according to any one of Embodiments 1 to 9, further comprising:

Embodiment 11. FIG.
A waste chamber formed in the substrate and through the mixing well and fluid through the channel formed in the substrate;
A fluid exit port formed in the substrate and through the mixing well and fluid through a channel formed in the substrate;
A first external actuable control valve disposed in the substrate and configured and arranged to selectively flow or stop fluid from the mixing well to the waste chamber;
A second externally operable control valve disposed in the substrate and configured to selectively flow fluid or stop fluid from the mixing well to the fluid ejection port;
11. The fluid sample processing cartridge of embodiments 1-10, further comprising:

Embodiment 12 FIG. The fluid sample processing cartridge of embodiment 11, further comprising a capture chamber disposed along a channel connecting the mixing well and the waste chamber.

Embodiment 13. FIG.
Closes when the pressure in the mixing well is not greater than the threshold pressure, and prevents fluid from flowing out of the mixing well and opens when the pressure in the mixing well rises above the threshold pressure A passive valve assembly configured and arranged to flow fluid from the mixing well;
A pressure port formed in the substrate and through the pressure with the passive valve assembly by a pressure conduit formed in the substrate, when the pressure port is closed, the pressure in the mixing well is at a threshold pressure that opens the passive valve assembly. A pressure port that can be reached and allows fluid to flow from the mixing well and when the pressure port is open, the pressure in the mixture cannot reach the threshold pressure and the passive valve assembly is closed; and
The fluid sample processing cartridge of embodiments 1-12, further comprising:

Embodiment 14 FIG. Associated with a deformable fluid chamber and connected to the associated deformable fluid chamber or including a bead held in a lance blister by a connectable and breakable septum upon application of an external compressive force 14. The fluid sample processing cartridge of any one of embodiments 1-13, further comprising a lance blister that ruptures, thereby pushing the beads through the breakable septum.

Embodiment 15. FIG. Embodiment 15. The fluid sample processing cartridge according to any one of Embodiments 1 to 14, further comprising an external flash that wraps at least a portion of the cartridge from the outside.

Embodiment 16. FIG. A plurality of deformable fluid chambers, each of the fluid chambers being one or more selected from the group consisting of a lysis buffer, a wash buffer, an oil, a rehydration buffer, a target capture bead, and a binding buffer. Embodiment 16. The fluid sample processing cartridge of any one of embodiments 1-15, comprising the material of:

Embodiment 17. FIG.
A first fluid exit port formed in the substrate and through the mixing well and fluid through a channel formed in the substrate;
A second fluid exit port formed in the substrate;
At least two deformable fluid chambers, one of the two deformable fluid chambers passing through the mixing well and fluid through a channel formed in the substrate; The other has at least two deformable fluid chambers through which fluid is communicated with the second fluid ejection port through a channel formed in a different substrate than the channel that leads the first fluid ejection port to the mixing well;
The fluid sample processing cartridge according to any one of Embodiments 1 to 10, further comprising:

Embodiment 18. FIG. The deformable fluid chamber through the mixing well and fluid includes a lysis buffer, wash buffer, target capture bead, or binding buffer, and the deformable fluid chamber through the second fluid ejection and fluid is oil 18. The fluid sample processing cartridge of embodiment 17, comprising a rehydration buffer.

Embodiment 19. FIG.
i) a substrate;
ii) a sample well formed in the substrate and configured to receive a volume of fluid sample;
iii) a closure configured to be selectively placed on the sample well;
iv) is supported on the substrate and retains the fluid therein when undeformed, ruptures upon ejection of an external compressive force, and discharges at least a portion of the fluid from the first fluid chamber. A first deformable fluid chamber configured to pass through the sample well and fluid through a channel formed in the substrate;
v) a mixing well formed in the substrate and passing the sample well and fluid through the channel formed in the substrate;
vi) a drive mixing device disposed in the mixing well and configured and arranged to mix the contents of the mixing well;
vii) a first fluid exit port formed in the substrate and through the mixing well and fluid through a channel formed in the substrate;
A) a sample preparation module;
B) a reaction module attached to the sample preparation module and configured to receive fluid from the sample preparation module via a fluid ejection port formed in the sample preparation module;
1) an upper surface;
2) a rising wall that at least partially surrounds the perimeter of the upper surface, is in contact with the surface of the sample preparation module to seal fluid, and forms an intervening space between the upper surface and the surface of the sample preparation module;
3) a sample chamber fluidly coupled to the first fluid exit port of the sample preparation module;
4) a reagent chamber;
5) a detection chamber;
I) an upper plate;
A fluid processing panel coupled to a lower surface of an upper plate and defining a reaction / processing space between the fluid processing panel and the upper plate, wherein the reaction / processing space is relative to the sample chamber, reaction chamber, and detection chamber. Ii) a fluid treatment panel; and
B) a reaction module;
A fluid sample processing cartridge comprising:

Embodiment 20. FIG. The sample chamber of the reaction module includes an inlet port where a fluid sample enters the sample chamber and includes a gap between the first fluid exit port of the sample preparation module and the inlet port of the sample chamber, the gap being an intervening space Embodiment 20. The fluid sample processing cartridge of embodiment 19, wherein the fluid sample processing cartridge is open to.

Embodiment 21. FIG. The fluid sample processing cartridge of embodiment 19, wherein the first fluid exit port of the sample preparation module comprises an outlet channel formed through the frustoconical nipple.

Embodiment 22. FIG. Embodiment 22. The fluid sample processing cartridge of any one of embodiments 19-21, wherein the reaction module further comprises an electronic sensor array disposed in each detection chamber.

Embodiment 23. FIG. The top plate of the reaction module further comprises one or more bubble traps, each bubble trap comprising a bubble capture hood that is open to the reaction / treatment space and a vent opening that is open to the interstitial space. The fluid sample processing cartridge according to any one of Embodiments 19 to 22.

Embodiment 24. FIG. Sample preparation module
Supported and undeformed on the substrate, retains the fluid therein and is configured to rupture and eject at least a portion of the fluid from the fluid chamber when an external compressive force is applied. A second deformable fluid chamber;
A second deformable fluid chamber and a second fluid exit port through which the fluid passes through a channel formed in the substrate and through a channel formed in the substrate;
The reaction and processing space is fluidly coupled to the second fluid exit port of the sample preparation module;
The fluid sample processing cartridge according to any one of Embodiments 19 to 23.

Embodiment 25. FIG. Mix wells
The surrounding walls and floor defining the well;
A fluid inlet snorkel that extends to one side of the peripheral wall that extends from the channel leading to the mixing well to the sample well and stops under the upper end of the peripheral wall;
A fluid sample processing cartridge according to any one of embodiments 19-24, comprising:

Embodiment 26. FIG. 26. The fluid sample processing cartridge of embodiment 25, wherein the fluid inlet snorkel extends to the outer surface of the peripheral wall and stops at an opening formed in the peripheral wall.

Embodiment 27. FIG. 27. The embodiment of embodiment 25 or 26, wherein the mixing well further comprises an exit port comprising one or more openings formed in the floor of the mix well, the floor being tapered downward toward the exit port. Fluid sample processing cartridge.

Embodiment 28. FIG. The drive mixing device is drivably meshed with a first impeller rotatably disposed within the mixing well and a meshing gear of the device into which the fluid sample processing cartridge is inserted, and when engaged by the meshing gear, the first A gear configured to rotate the impeller of the
A fluid sample processing cartridge according to any one of embodiments 19-25, comprising:

Embodiment 29. FIG. Sample preparation module
A lysis chamber comprising a plurality of lysis beads, formed in a substrate and disposed along a channel connecting a mixing well and a sample well, wherein fluid flowing from the sample well to the mixing well flows through the lysis chamber A dissolution chamber;
A bead mixer disposed and arranged at least partially within the lysis chamber and configured to agitate the lysis beads and the fluid flowing through the lysis chamber;
The fluid sample processing cartridge of any one of embodiments 19-28, further comprising:

Embodiment 30. FIG.
A first optical interface comprising an enlarged portion of a channel connecting the lysis chamber to the sample well;
A second optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the mixing well;
30. The fluid sample processing cartridge of embodiment 29, further comprising:

Embodiment 31. FIG. The bead mixer
A motor mounted on the substrate;
A second impeller disposed in the dissolution chamber and mounted on the output shaft of the motor;
31. A fluid sample processing cartridge according to embodiment 29 or 30, comprising:

Embodiment 32. FIG. The lysis chamber includes a fluid inlet and a fluid outlet, further comprising a mesh filter disposed on each of the fluid inlet and the fluid outlet and configured to retain lysis beads in the lysis chamber. 31. The fluid sample processing cartridge according to any one of 31.

Embodiment 33. FIG. Sample preparation module
A pressure port formed in the substrate and configured to couple the substrate to an external fluid pressure source;
A channel formed in the substrate connecting the pressure port to the sample well;
The fluid sample processing cartridge according to any one of embodiments 19 to 32, further comprising:

Embodiment 34. FIG. Sample preparation module
A waste chamber formed in the substrate and through the mixing well and fluid through the channel formed in the substrate;
A first externally operable control valve disposed within the substrate and configured and arranged to selectively flow fluid from the mixing well to the waste chamber or to prevent fluid flow;
A second externally operable control valve disposed in the substrate and configured and arranged to selectively flow fluid from the mixing well to the exit port or to prevent fluid flow;
34. The fluid sample processing cartridge of embodiments 19-33, further comprising:

Embodiment 35. FIG. 35. The fluid sample processing cartridge of embodiment 34, wherein the sample preparation module further comprises a capture chamber disposed along a channel connecting the mixing well and the waste chamber.

Embodiment 36. FIG. Sample preparation module
Placed in the substrate and closed if the pressure in the mixing well is not greater than the threshold pressure, blocking fluid flow from the mixing well and opening if the pressure in the mixing well rises above the threshold pressure. A passive valve assembly configured and arranged to flow fluid from the mixing well; and
A pressure port formed in the substrate and through the pressure with the passive valve assembly by a pressure conduit formed in the substrate, the pressure in the mixing well when the pressure port is closed, the threshold at which the passive valve assembly will open A pressure port capable of reaching pressure, allowing fluid to flow from the mixing well, and when the pressure port is opened, the pressure in the mixing well cannot reach the threshold pressure and the passive valve assembly is closed; and
36. The fluid sample processing cartridge of embodiments 19-35, further comprising:

Embodiment 37. FIG. The sample preparation module includes a bead associated with the deformable fluid chamber and connected to the associated deformable fluid chamber or held in the lance blister by a connectable and breakable septum; 37. The device of any one of embodiments 19-36, further comprising a lance blister configured to rupture when applied with an external compressive force, thereby pushing the bead through the breakable septum. Fluid sample processing cartridge.

Embodiment 38. FIG. 38. The fluid sample processing cartridge of any one of embodiments 19 to 37, further comprising an external flash that wraps at least a portion of the cartridge from the outside.

Embodiment 39. FIG. The sample preparation module further comprises a plurality of deformable fluid chambers, each of the fluid chambers from the group consisting of a lysis buffer, a wash buffer, an oil, a rehydration buffer, a target capture bead, and a binding buffer. 39. A fluid sample processing cartridge according to any one of embodiments 19-38, comprising a selected material.

Embodiment 40. FIG. Deformable, supported on a flat substrate and held in an undeformed state, configured to hold the fluid therein and to rupture and eject at least a portion of the fluid from the fluid chamber when an external compressive force is applied An apparatus configured to process a fluid sample processing cartridge including a fluid chamber comprising:
A cartridge carriage assembly configured to receive and hold a fluid sample processing cartridge inserted into the device;
Between a first position adjacent to the cartridge carriage assembly and not in operative contact with a cartridge carried in the cartridge carriage assembly and a second position in operative contact with a cartridge carried in the cartridge carriage assembly A heating and control assembly configured for movement relative to the cartridge carriage assembly;
For movement relative to the cartridge, independent of the heating and control assembly, between a first position that does not substantially apply a magnetic force to the cartridge and a second position that applies a magnetic force to a corresponding individual portion of the cartridge. One or more movable magnet assemblies each mounted;
A cartridge carriage assembly configured for powered movement, wherein the powered movement of the cam block assembly is between a first position of the heating and control assembly and a second position of the heating and control assembly. Operatively coupled to the heating and control assembly to convert the powered movement of the cam block assembly to a first position of the magnet assembly and a second of the magnet assembly. A cam block assembly operatively coupled to one or more movable magnet assemblies for converting the movement of each magnet assembly relative to the cartridge carriage assembly between positions;
e) a deformable chamber compression assembly configured to selectively apply an external compressive force to the deformable fluid chamber, rupture the deformable chamber, and discharge at least a portion of the fluid from the fluid chamber;
A device comprising:

Embodiment 41. FIG. The heating and control assembly
One or more heater assemblies configured to apply a thermal gradient to a corresponding discrete portion of the cartridge when the heating and control assembly is in the second position;
A connector board including one or more electrical connector elements configured to enable an electrical connection between the device and the cartridge when the heating and control assembly is in the second position;
41. The apparatus of embodiment 40, comprising:

Embodiment 42. FIG. The deformable chamber compression assembly is
A cam follower plate configured for movement supplied with power in a first direction substantially parallel to the plane of the substrate;
A compression mechanism associated with the deformable chamber of the cartridge and configured to apply a force to compress the chamber against the substrate by movement in a second direction having a component substantially perpendicular to the plane of the substrate; With
Embodiment 40, wherein the cam follower plate translates the movement of the cam follower plate in the first direction into the movement of the compression mechanism in the second direction, thereby operably coupling with a compression mechanism that applies an external compression force to the chamber. Or the apparatus as described in 41.

Embodiment 43. FIG. An embodiment further comprising an air pump and an air port connected to the air pump, wherein the air port is configured to couple the air pump to the pressure port of the fluid sample processing cartridge when the cartridge is inserted into the device. The apparatus according to any one of 40 to 42.

Embodiment 44. FIG. 44. The apparatus according to any one of embodiments 40-43, further comprising a photodetector configured to detect fluid flow through a portion of the fluid sample processing cartridge.

Embodiment 45. FIG. The fluid sample processing cartridge includes a drive mixing device including a drive gear, the device includes a powered gear, a first position where the driving gear does not engage the drive gear of the drive mixing device, and the driving gear. Further comprising a mixing motor assembly that is operatively engaged with the drive gear and movable between a second position for actuating the drive mixing device, the cam block assembly being powered by the cam block assembly Embodiments 40-44 for operatively coupling a mixing motor assembly to translate movement of the mixing motor assembly between a first position of the mixing motor assembly and a second position of the mixing motor assembly The apparatus of any one of these.

Embodiment 46. FIG.
With fans,
A cooling duct configured to flow air from the fan to a portion of the heater assembly;
46. The apparatus of any one of embodiments 41-45, further comprising a heater cooling assembly comprising:

Embodiment 47. The cartridge carriage assembly
A cartridge holder configured to hold a cartridge to be inserted;
A cartridge latch biased to a cartridge latch position and configured to latch a cartridge inserted into the cartridge holder and hold the cartridge within the cartridge holder;
A cartridge ejection mechanism configured to automatically push the cartridge at least partially out of the cartridge holder when the cartridge latch is released from the cartridge latch position;
47. The apparatus according to any one of embodiments 40-46, comprising:

Embodiment 48. FIG. The heating and control assembly includes a support plate on which one or more heater assemblies and a connector board are supported, the support plate preventing horizontal movement of the support plate but allowing vertical movement of the support plate. 48. The apparatus of any one of embodiments 41-47, mounted in a constrained configuration that allows the heating and control assembly to move between its first position and a second position.

Embodiment 49. One of the heater assemblies of the heating and control assembly comprises a resistance heating element attached to the connector board and a heat spreader comprising a thermally conductive material that is thermally coupled to the resistance heating element. 49. The apparatus according to any one of 48.

Embodiment 50. FIG. One of the heater assemblies in the heating and control assembly is
A thermoelectric element;
A heat spreader comprising a thermally conductive material thermally coupled to the thermoelectric element;
A heat sink including a panel in thermal contact with the thermoelectric element and the plurality of heat dissipating rods;
50. The apparatus according to any one of embodiments 41-49, comprising:

Embodiment 51. FIG. The apparatus of any one of embodiments 41-50, wherein the electrical connector elements of the connector board of the heating and control assembly comprise a plurality of connector pin arrays, each connector pin array comprising a plurality of pogo pins.

Embodiment 52. FIG. One of the movable magnet assemblies is
A magnet holder mounted on the spindle and adapted to rotate about the spindle between a first position and a second position of the magnet assembly;
A magnet supported by a magnet holder;
An actuator bracket extending from the magnet holder;
A torsion spring configured to bias the magnet holder to a rotational position corresponding to the first position of the magnet assembly;
52. The apparatus according to any one of embodiments 40-51, comprising:

Embodiment 53. FIG. One of the movable magnet assemblies is
A magnet holder frame mounted on the spindle and rotatable about the spindle between a first position and a second position of the magnet assembly;
A magnet array disposed within the magnet holder frame;
A converging magnet disposed in an opening formed in the magnet holder frame and configured to converge the magnetic force of the magnet array;
An actuator bracket extending from the magnet holder frame;
A torsion spring configured to bias the magnet holder frame to a rotational position corresponding to a first position of the magnet assembly;
54. The apparatus of any one of embodiments 40-52, comprising:

Embodiment 54. FIG. The cam block assembly is operably coupled to each movable magnet assembly by a magnet actuator that is partially coupled to the cam block assembly and is movable by a powered movement of the cam block assembly. Includes a tab configured to engage with an actuator bracket of each magnet assembly to move with the cam block assembly to cause a corresponding rotation of the magnet assembly from a first position to a second position. 54. Apparatus according to embodiment 52 or 53.

Embodiment 55. FIG. The cam block assembly
A cam frame,
A cam block motor coupled to the cam frame and configured to enable powered movement of the cam frame;
First and second cam rails attached to the cam frame, each of the cam rails having two cam slots, wherein the cam block assembly is heated by a cam follower extending from the heating and control assembly to the cam slot. A cam operatively coupled to the control assembly, wherein the movement of the cam frame and cam rail relative to the heating and control assembly corresponds to a corresponding relative movement between the cam follower and cam slot to a first position of the heating and control assembly Causing a movement of the cam follower between a first segment of each of the slots and a respective second segment of the cam slot corresponding to a second position of the heating and control assembly; First and second cam rails;
55. The apparatus according to any one of embodiments 40-54, comprising:

Embodiment 56. FIG. The cam frame
A first longitudinal circle extending along one side of the heating and control assembly;
A second longitudinal circle extending along the opposing surface of the heating and control assembly;
A cross circular material extending between first and second long circular members, each cam rail being attached to one of the first and second long circular members. When,
56. The apparatus according to embodiment 55, comprising:

Embodiment 57. FIG. The compression mechanism of the deformable chamber compression assembly is:
A cam arm having a cam surface and mounted so as to pivot about one end of the cam arm;
A compression pad disposed at the opposite end of the cam arm, the cam arm applying a compression force to the first position where the compression pad does not contact the deformable chamber associated with the compression pad and the deformable chamber associated with the compression pad. A compression pad that pivots at least partially between a second position to rupture the chamber;
57. The apparatus according to any one of embodiments 42-56, comprising:

Embodiment 58. FIG. The deformable chamber compression assembly further includes a cam arm plate, and the cam arm of the compression mechanism is pivotally mounted within a slot formed in the cam arm plate for pivoting of the cam arm relative to the cam arm plate. And the cam surface of the cam arm protrudes out of the slot above the surface of the cam arm plate, and the cam follower plate pivots the cam arm from the first position to the second position. 58. The apparatus of embodiment 57, wherein the plate is operatively coupled to the compression mechanism by a cam follower element of the cam follower plate that engages a cam surface of the compression mechanism during plate movement.

Embodiment 59. The cam follower plate includes a cam groove that receives the cam surface of the cam arm protruding above the surface of the cam arm plate. The cam follower element moves the cam follower plate relative to the cam arm plate to move the cam arm from the first position. 59. The apparatus of embodiment 58, comprising a follower ridge disposed in a cam groove in contact with the cam surface to pivot to the second position.

Embodiment 60. FIG. Each comprises a plurality of compression mechanisms comprising a cam arm plate and a cam arm movably mounted in a slot formed in the cam arm surface, the cam follower plate comprising a plurality of cam grooves, each cam groove comprising at least Associated with one compression mechanism, each cam groove is associated with the cam follower plate as it moves relative to the cam arm plate to pivot the cam arm of the associated compression mechanism from the first position to the second position. 60. The apparatus of embodiment 59, comprising a follower ridge disposed in a cam groove that contacts a cam surface of the compression mechanism.

Embodiment 61. FIG. The sample processing cartridge includes a plurality of deformable fluid chambers, the deformable chamber compression assembly includes a plurality of compression mechanisms, each compression mechanism associated with one of the deformable fluid chambers, and the cam follower plate To convert the movement of the cam follower plate in the first direction to the respective movement of the compression mechanism in the second direction, thereby applying an external compressive force to each of the associated chambers in a specific sequence 61. The apparatus of any one of embodiments 42-60, operably coupled to a compression mechanism.

Embodiment 62. FIG. The fluid sample processing cartridge allows fluid flow through the valve when not actuated externally, and prevents fluid flow through the valve when actuated externally. A second direction having a component substantially perpendicular to the plane of the substrate relative to the externally operable control valve of the sample processing cartridge. Further comprising a valve actuator compression mechanism configured to actuate an associated externally actuatable control valve upon movement of the cam follower plate, wherein the cam follower plate moves the cam follower plate in the first direction. To convert the movement of the actuator compression mechanism, thereby activating the associated external actuatable control valve Operatively coupled to the valve actuator compression mechanism, apparatus according to any one of embodiments 40-61.

  Although the subject matter of this disclosure has been described and illustrated in considerable detail with reference to certain exemplary embodiments, including various combinations and subcombinations of features, those skilled in the art will recognize other embodiments, variations, and These modifications will be readily understood as falling within the scope of the present invention. Furthermore, the description of such embodiments, combinations, and subcombinations is not intended to convey that the invention requires features or combinations of features other than those expressly recited in a claim. Accordingly, the scope of this disclosure is intended to include all modifications and variations that fall within the spirit and scope of the following appended claims.

Claims (62)

A substrate,
A sample well formed in the substrate and configured to receive a volume of fluid sample;
A closure configured to be selectively placed on the sample well;
In a state where it is supported on the substrate and is not deformed, the fluid is retained therein, and when an external compressive force is applied, it ruptures and discharges at least a portion of the fluid from the fluid chamber. A deformable fluid chamber configured and in fluid communication with the sample well through a channel formed in the substrate;
The sample well is formed from the channel, which is formed in the substrate, passes the sample well and fluid through a channel formed in the substrate, communicates with a first peripheral wall and a first floor defining the well, and a mixing well. A mixing well comprising: a fluid inlet snorkel extending to one side of the first peripheral wall extending to and stopping below an upper end of the first peripheral wall;
A drive mixing device disposed in the mixing well and configured and arranged to mix the contents of the mixing well;
A fluid sample processing cartridge comprising:   The fluid sample processing cartridge of claim 1, wherein the fluid inlet snorkel extends on an outer surface of the first peripheral wall and stops at an opening formed in the first peripheral wall.   The sample well includes a second peripheral wall and a second floor defining a well; a fluid inlet snorkel that extends over one surface of the second peripheral wall and stops under an upper end of the second peripheral wall; A fluid sample processing cartridge according to claim 1 or 2, comprising:   The mixing well further comprises an exit port with one or more openings formed in the second floor, the floor being tapered downwardly toward the exit port. The fluid sample processing cartridge according to claim 1.   The drive mixing device meshes with a first impeller disposed rotatably in the mixing well so as to be driven by a meshing gear of a device into which the fluid sample processing cartridge is inserted, and the meshing gear is A fluid sample processing cartridge according to any one of claims 1 to 4, further comprising a gear configured to rotate the first impeller when engaged. A lysis chamber comprising a plurality of lysis beads, formed on the substrate and disposed along the channel connecting the mixing well and the sample well, wherein the fluid flowing from the sample well to the mixing well comprises: Flowing through the dissolution chamber;
6. The bead mixer of any of claims 1-5, further comprising a bead mixer disposed at least partially within the lysis chamber and configured and arranged to agitate the lysis beads and fluid flowing through the lysis chamber. The fluid sample processing cartridge according to claim 1. A first optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the sample well;
A second optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the mixing well;
The fluid sample processing cartridge of claim 6, further comprising: The bead mixer is
A motor mounted in the substrate;
A second impeller disposed in the dissolution chamber and mounted on the output shaft of the motor;
A fluid sample processing cartridge according to claim 6 or 7.   The lysis chamber includes a fluid inlet and a fluid outlet, further comprising a mesh filter disposed on each of the fluid inlet and the fluid outlet and configured to retain the lysis beads in the lysis chamber; The fluid sample processing cartridge according to any one of claims 6 to 8. A pressure port formed in the substrate and configured to couple the substrate to an external fluid pressure source;
A channel formed in the substrate connecting the pressure port to the sample well;
The fluid sample processing cartridge according to claim 1, further comprising: A waste chamber formed in the substrate and passing the mixing well and fluid through a channel formed in the substrate;
A fluid exit port formed in the substrate and through the mixing well and fluid through a channel formed in the substrate;
A first externally operable control valve disposed in the substrate and configured and arranged to selectively flow fluid to or from the waste chamber from the mixing well; and
A second externally operable control valve disposed within the substrate and configured to selectively flow fluid or stop fluid from the mixing well to the fluid ejection port;
The fluid sample processing cartridge according to claim 1, further comprising:   The fluid sample processing cartridge of claim 11, further comprising a capture chamber disposed along a channel connecting the mixing well and the waste chamber. Closed when the pressure in the mixing well is not greater than a threshold pressure, and no fluid flows from the mixing well, the pressure in the mixing well rises above the threshold pressure A passive valve assembly configured and arranged to open and flow fluid from the mixing well when
A pressure port formed in the substrate and through the passive valve assembly and pressure by a pressure conduit formed in the substrate, wherein the pressure in the mixing well is reduced when the pressure port is closed. The threshold pressure that can open the assembly can be reached, allowing fluid to flow from the mixing well, and when the pressure port is open, the pressure in the mixing cannot reach the threshold pressure; A pressure port for closing the passive valve assembly; and
The fluid sample processing cartridge according to claim 1, further comprising:   An external compressive force associated with the deformable fluid chamber, comprising beads held in the lance blister by a connectable and breakable septum connected to the associated deformable fluid chamber; 14. A fluid sample processing cartridge according to any one of the preceding claims, further comprising a lance blister configured to rupture upon application of the pressure, thereby pushing the beads through the breakable septum. .   The fluid sample processing cartridge according to claim 1, further comprising an external flash that wraps at least a part of the cartridge from the outside.   A plurality of deformable fluid chambers, each of which is selected from the group consisting of a lysis buffer, a wash buffer, an oil, a rehydration buffer, a target capture bead, and a binding buffer 1 The fluid sample processing cartridge according to any one of claims 1 to 15, comprising the above substances. A first fluid exit port formed in the substrate and through the mixing well and fluid through a channel formed in the substrate;
A second fluid exit port formed in the substrate;
At least two deformable fluid chambers, wherein one of the two deformable fluid chambers passes the mixing well and fluid through a channel formed in the substrate. The other of the fluid chambers passes fluid through the second fluid ejection port through a channel formed in the substrate different from the channel leading the first fluid ejection port to the mixing well. Two deformable fluid chambers;
The fluid sample processing cartridge according to claim 1, further comprising:   The deformable fluid chamber through the mixing well and fluid includes a lysis buffer, a wash buffer, a target capture bead, or a binding buffer, and the deformable fluid through the second fluid ejection and fluid. The fluid sample processing cartridge of claim 17, wherein the chamber comprises oil or a rehydration buffer. i) a substrate;
ii) a sample well formed in the substrate and configured to receive a volume of fluid;
iii) a closure configured to be selectively placed on the sample well;
iv) is supported on the substrate and retains the fluid therein when undeformed, ruptures upon application of an external compressive force, and discharges at least a portion of the fluid from the first fluid chamber A first deformable fluid chamber that is in fluid communication with the sample well through a channel formed in the substrate;
v) a mixing well formed in the substrate and passing the sample well and fluid through a channel formed in the substrate;
vi) a drive mixing device disposed in the mixing well and configured and arranged to mix the contents of the mixing well;
vii) a first fluid exit port formed in the substrate and through the mixing well and fluid through a channel formed in the substrate;
A) a sample preparation module;
B) a reaction module attached to the sample preparation module and configured to receive fluid from the sample preparation module via the fluid exit port formed in the sample preparation module;
1) an upper surface;
2) at least partially surrounding the periphery of the upper surface, in contact with the surface of the sample preparation module so as to seal fluid, and forming an intervening space between the upper surface and the surface of the sample preparation module The rising wall,
3) a sample chamber that is fluidly coupled to the first fluid exit port of the sample preparation module;
4) a reagent chamber;
5) a detection chamber;
I) an upper plate;
A fluid processing panel coupled to a lower surface of the upper plate and defining a reaction / processing space between the fluid processing panel and the upper plate, wherein the reaction / processing space includes the sample chamber, the reaction chamber, And ii) a fluid treatment panel that is open to or open to the detection chamber;
B) a reaction module;
A fluid sample processing cartridge comprising:   The sample chamber of the reaction module includes an inlet port where a fluid sample enters the sample chamber, with a gap between the first fluid exit port of the sample preparation module and the inlet port of the sample chamber. 20. The fluid sample processing cartridge of claim 19, wherein the gap is open to an intervening space.   20. A fluid sample processing cartridge according to claim 19, wherein the first fluid exit port of the sample preparation module comprises an outlet channel formed through a frustoconical nipple.   The fluid sample processing cartridge according to any one of claims 19 to 21, wherein the reaction module further comprises an electronic sensor array disposed in each detection chamber.   The upper plate of the reaction module further includes one or more bubble traps, each bubble trap having a bubble trapping hood that is open to the reaction / processing space, and a vent opening that is open to the intervening space. The fluid sample processing cartridge according to any one of claims 19 to 22, comprising a portion. The sample preparation module includes:
In a state where the fluid is supported on the substrate and not deformed, the fluid is held inside, and when an external compressive force is applied, the fluid is ruptured, and at least a part of the fluid is discharged from the fluid chamber. A second deformable fluid chamber,
A second fluid exit port that is supported by the substrate and through the channel formed in the substrate and through which the second deformable fluid chamber passes fluid; and
The reaction and processing space is fluidly coupled to the second fluid exit port of the sample preparation module;
24. A fluid sample processing cartridge according to any one of claims 19 to 23. The mixing well is
The surrounding walls and floor defining the well;
A fluid inlet snorkel extending to one side of the peripheral wall extending from the channel leading to the mixing well to the sample well and stopping under the upper end of the peripheral wall;
25. A fluid sample processing cartridge according to any one of claims 19 to 24.   26. The fluid sample processing cartridge of claim 25, wherein the fluid inlet snorkel extends to an outer surface of the peripheral wall and stops at an opening formed in the peripheral wall.   26. The mixing well further comprises an exit port comprising one or more openings formed in the floor of the mix well, the floor being tapered downward toward the exit port. The fluid sample processing cartridge according to claim 26. The drive mixing device is engaged with a first impeller rotatably arranged in the mixing well and a meshing gear of a device into which the fluid sample processing cartridge is inserted, and meshed by the meshing gear. A gear configured to rotate the first impeller;
26. A fluid sample processing cartridge according to any one of claims 19 to 25. The sample preparation module includes:
A lysis chamber comprising a plurality of lysis beads, formed on the substrate and disposed along the channel connecting the mixing well and the sample well, wherein the fluid flowing from the sample well to the mixing well is A dissolution chamber flowing through the dissolution chamber;
A bead mixer disposed and disposed at least partially within the lysis chamber and configured to agitate the lysis beads and fluid flowing through the lysis chamber;
The fluid sample processing cartridge according to any one of claims 19 to 28, further comprising: A first optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the sample well;
A second optical interface comprising an enlarged portion of the channel connecting the lysis chamber to the mixing well;
30. The fluid sample processing cartridge of claim 29, further comprising: The bead mixer is
A motor mounted in the substrate;
A second impeller disposed in the dissolution chamber and mounted on the output shaft of the motor;
31. A fluid sample processing cartridge according to claim 29 or claim 30, comprising:   The lysis chamber includes a fluid inlet and a fluid outlet, and is disposed on each of the fluid inlet and the fluid outlet, and is configured to retain the lysis beads in the lysis chamber; and The fluid sample processing cartridge according to any one of claims 29 to 31, further comprising: The sample preparation module includes:
A pressure port formed in the substrate and configured to couple the substrate to an external fluid pressure source;
A channel formed in the substrate connecting the pressure port to the sample well;
The fluid sample processing cartridge according to claim 19, further comprising: The sample preparation module includes:
A waste chamber formed in the substrate and through the mixing well and fluid through a channel formed in the substrate;
A first externally operable control valve disposed in the substrate and configured and arranged to selectively flow fluid from the mixing well to the waste chamber or to stop fluid flow;
A second externally operable control valve disposed within the substrate and configured and arranged to selectively flow fluid from the mixing well to the exit port or to stop fluid flow;
34. The fluid sample processing cartridge according to any one of claims 19 to 33, further comprising:   35. The fluid sample processing cartridge of claim 34, wherein the sample preparation module further comprises a capture chamber disposed along a channel connecting the mixing well and the waste chamber. The sample preparation module includes:
If placed in the substrate and the pressure in the mixing well is not greater than a threshold pressure, it is closed to prevent fluid flow from the mixing well, and the pressure in the mixing well exceeds the threshold pressure. A passive valve assembly configured and arranged to open and flow fluid from the mixing well when rising;
A pressure port formed in the substrate and through which pressure is passed through the passive valve assembly by a pressure conduit formed in the substrate, wherein when the pressure port is closed, the pressure in the mixing well is reduced by the passive valve assembly. When the pressure port is opened, the pressure in the mixing well cannot reach the threshold pressure, and when the pressure port is opened, the threshold pressure can be reached. A pressure port where the passive valve assembly is closed; and
36. The fluid sample processing cartridge according to any one of claims 19 to 35, further comprising:   The sample preparation module is associated with the deformable fluid chamber and is connected to the associated deformable fluid chamber or a bead held in a lance blister by a connectable and breakable septum 37. A lance blister further comprising a lance blister configured to rupture when applied with an external compressive force, thereby pushing the bead through the breakable septum. 2. The fluid sample processing cartridge according to item 1.   38. The fluid sample processing cartridge according to any one of claims 19 to 37, further comprising an external flash that wraps at least a portion of the cartridge from the outside.   The sample preparation module further comprises a plurality of deformable fluid chambers, each of the fluid chambers comprising a lysis buffer, a wash buffer, an oil, a rehydration buffer, a target capture bead, and a binding buffer. 39. A fluid sample processing cartridge according to any one of claims 19 to 38, comprising a material selected from the group. A deformation that is supported on a flat substrate and is not deformed, retains the fluid therein, and ruptures and discharges at least a portion of the fluid from the fluid chamber when an external compressive force is applied. An apparatus configured to process a fluid sample processing cartridge comprising a possible fluid chamber comprising:
A cartridge carriage assembly configured to receive and hold a fluid sample processing cartridge inserted into the device;
A first position adjacent to the cartridge carriage assembly and not in operative contact with the cartridge carried in the cartridge carriage assembly; and a second position in operative contact with the cartridge carried in the cartridge carriage assembly. A heating and control assembly configured for movement relative to the cartridge carriage assembly between
With respect to the cartridge, independent of the heating and control assembly, between a first position that does not substantially apply a magnetic force to the cartridge and a second position that applies a magnetic force to a corresponding discrete portion of the cartridge. One or more movable magnet assemblies each mounted for movement;
Configured for powered movement, the powered movement of the cam block assembly between the first position of the heating and control assembly and the second position of the heating and control assembly , Operatively coupled to the heating and control assembly for converting the movement of the heating and control assembly relative to the cartridge carriage assembly, and the powered movement of the cam block assembly is coupled to the first of the magnet assembly. A cam block operably coupled to the one or more movable magnet assemblies for converting the movement of each magnet assembly relative to the cartridge carriage assembly between a position of 1 and the second position of the magnet assembly. Assembly,
e) A deformable chamber configured to selectively apply an external compressive force to the deformable fluid chamber to rupture the deformable chamber and to discharge at least a portion of the fluid from the fluid chamber. A compression assembly;
A device comprising: The heating and control assembly includes:
One or more heater assemblies configured to apply a thermal gradient to a corresponding discrete portion of the cartridge when the heating and control assembly is in the second position;
A connector board including one or more electrical connector elements configured to enable an electrical connection between the device and the cartridge when the heating and control assembly is in the second position;
41. The apparatus of claim 40, comprising: The deformable chamber compression assembly comprises:
A cam follower plate configured for movement supplied with power in a first direction substantially parallel to the plane of the substrate;
The cartridge is configured to apply a force to compress the chamber relative to the substrate by movement in a second direction having a component substantially perpendicular to the plane of the substrate relative to the deformable chamber. A compression mechanism,
The cam follower plate converts the movement of the cam follower plate in the first direction into movement of the compression mechanism in the second direction, thereby applying the external compression force to the chamber. 42. An apparatus as claimed in claim 40 or claim 41, wherein the apparatus is operably coupled to the apparatus.   And an air port connected to the air pump, the air port coupling the air pump to a pressure port of the fluid sample processing cartridge when the cartridge is inserted into the device. 43. Apparatus according to any one of claims 40 to 42 configured.   44. The apparatus according to any one of claims 40 to 43, further comprising a photodetector configured to detect fluid flow through a portion of the fluid sample processing cartridge.   The fluid sample processing cartridge includes a drive mixing device that includes a drive gear, the device includes a drive gear that is powered, and a driving gear does not engage the drive gear of the drive mixing device. And a driving motor assembly that is operatively engaged with the drive gear and movable between a second position for operating the drive mixing device, the cam block assembly comprising the cam block assembly To convert the powered movement of the block assembly into movement of the mixing motor assembly between the first position of the mixing motor assembly and the second position of the mixing motor assembly. 45. Apparatus according to any one of claims 40 to 44, operatively coupled to a mixing motor assembly. With fans,
A cooling duct configured to flow air from the fan to a portion of the heater assembly;
46. The apparatus of any one of claims 41 to 45, further comprising a heater cooling assembly comprising: The cartridge carriage assembly includes
A cartridge holder configured to hold a cartridge to be inserted;
A cartridge latch biased to a cartridge latch position, configured to latch a cartridge inserted into the cartridge holder, and to hold the cartridge within the cartridge holder;
A cartridge ejection mechanism configured to automatically push the cartridge at least partially out of the cartridge holder when the cartridge latch is released from a cartridge latch position;
47. Apparatus according to any one of claims 40 to 46, comprising:   The heating and control assembly includes a support plate on which the one or more heater assemblies and the connector board are supported. The support plate prevents horizontal movement of the support plate, but is perpendicular to the support plate. 48. Any one of claims 41 to 47, mounted in a constrained configuration that permits directional movement to allow the heating and control assembly to move between its first and second positions. The device described in 1.   One of the heater assemblies of the heating and control assembly comprises a resistance heating element attached to the connector board and a heat spreader including a thermally conductive material that is thermally coupled to the resistance heating element. Item 49. The apparatus according to any one of Items 41 to 48. One of the heater assemblies of the heating and control assembly is:
A thermoelectric element;
A heat spreader comprising a thermally conductive material thermally coupled to the thermoelectric element;
A heat sink including a panel in thermal contact with the thermoelectric element and the plurality of heat dissipating rods;
50. Apparatus according to any one of claims 41 to 49, comprising:   51. The apparatus according to any one of claims 41 to 50, wherein the electrical connector element of the connector board of the heating and control assembly comprises a plurality of connector pin arrays, each connector pin array comprising a plurality of pogo pins. . One of the movable magnet assemblies is:
A magnet holder mounted on a spindle and adapted to rotate about the spindle between the first position and the second position of the magnet assembly;
A magnet supported by the magnet holder;
An actuator bracket extending from the magnet holder;
A torsion spring configured to bias the magnet holder to a rotational position corresponding to the first position of the magnet assembly;
52. The apparatus according to any one of claims 40 to 51, comprising: One of the movable magnet assemblies is:
A magnet holder frame mounted on a spindle and rotatable about the spindle between the first position and the second position of the magnet assembly;
A magnet array disposed within the magnet holder frame;
A converging magnet arranged in an opening formed in the magnet holder frame and configured to converge the magnetic force of the magnet array;
An actuator bracket extending from the magnet holder frame;
A torsion spring configured to bias the magnet holder frame to a rotational position corresponding to the first position of the magnet assembly;
53. Apparatus according to any one of claims 40 to 52, comprising:   The cam block assembly is operably coupled to each movable magnet assembly by a magnet actuator that is partially coupled to the cam block assembly and is movable by a powered movement of the cam block assembly. The magnet actuator can engage with the actuator bracket of each magnet assembly such that the magnet actuator moves with the cam block assembly to cause a corresponding rotation of the magnet assembly from the first position to the second position. 54. Apparatus according to claim 52 or claim 53, comprising a tab configured as such. The cam block assembly includes:
A cam frame,
A cam block motor coupled to the cam frame and configured to enable powered movement of the cam frame;
First and second cam rails attached to the cam frame, each of the cam rails having two cam slots, the cam block assembly extending from the heating and control assembly to the cam slot. A cam follower is operably coupled to the heating and control assembly such that movement of the cam frame and the cam rail relative to the heating and control assembly results in a corresponding relative movement between the cam follower and the cam slot. A first segment of each of the cam slots corresponding to the first position of the control assembly and a second of each of the cam slots corresponding to the second position of the heating and control assembly. Causing the cam follower to move between the first and second segments; And a second cam rail,
55. Apparatus according to any one of claims 40 to 54, comprising: The cam frame is
A first elongate circular member extending along one surface of the heating and control assembly;
A second longitudinal circle extending along opposing surfaces of the heating and control assembly;
A cross member extending between the first and second longitudinal members, each cam rail being attached to one of the first and second longitudinal members. With circular material,
56. The apparatus of claim 55, comprising: The compression mechanism of the deformable chamber compression assembly comprises:
A cam arm having a cam surface and mounted so as to pivot about one end of the cam arm;
A compression pad disposed at an opposite end of the cam arm, the cam arm having a first position where the compression pad does not contact the associated deformable chamber, and the deformable chamber to which the compression pad is associated; A compression pad that pivots between a second position that applies a compression force to and at least partially ruptures the chamber;
57. The apparatus according to any one of claims 42 to 56, comprising:   The deformable chamber compression assembly further comprises a cam arm plate, the cam arm of the compression mechanism pivoting into a slot formed in the cam arm plate for pivoting of the cam arm relative to the cam arm plate. Operatively mounted, the cam surface of the cam arm protrudes out of the slot over the surface of the cam arm plate, and the cam follower plate moves the cam arm from a first position to a second position. 58. The cam follower element of the cam follower plate that operatively couples to the cam surface of the compression mechanism during movement of the cam follower plate relative to the cam arm plate to pivot to the compression mechanism. The device described in 1.   The cam follower plate includes a cam groove that receives the cam surface of the cam arm protruding above the surface of the cam arm plate, and the cam follower element moves the cam follower plate relative to the cam arm plate, 59. The apparatus of claim 58, comprising a follower ridge disposed in the cam groove that contacts the cam surface when pivoting the cam arm from a first position to a second position.   Each comprises a plurality of compression mechanisms comprising cam arms that are pivotally mounted in slots formed in the cam arm plate and cam arm surface, the cam follower plate comprising a plurality of cam grooves, each cam groove comprising: The cam follower plate is associated with at least one of the compression mechanisms, and each cam groove pivots the cam arm of the associated compression mechanism from a first position to a second position. 60. The apparatus of claim 59, comprising a follower ridge disposed in the cam groove that contacts the cam surface of the associated compression mechanism when moving relative to the cam.   The sample processing cartridge includes a plurality of deformable fluid chambers, the deformable chamber compression assembly comprises a plurality of compression mechanisms, each compression mechanism being associated with one of the deformable fluid chambers, The cam follower plate converts the movement of the cam follower plate in the first direction into a respective movement of the compression mechanism in the second direction, thereby each of the associated chambers in a particular sequence. 61. The apparatus according to any one of claims 42 to 60, wherein the apparatus is operatively coupled to the compression mechanism to apply an external compression force to the compression mechanism. The fluid sample processing cartridge allows fluid to flow through the valve when not actuated externally, and prevents fluid from flowing through the valve when actuated externally, An externally operable control valve configured to selectively control fluid flow, wherein the apparatus is associated with the externally operable control valve of the sample processing cartridge and is substantially perpendicular to the plane of the substrate. Further comprising a valve actuator compression mechanism configured to actuate the associated externally actuatable control valve by movement in a second direction having a component, wherein the cam follower plate includes the cam follower in the first direction. Converting the movement of the plate into the movement of the valve actuator compression mechanism in the second direction; , To actuate the associated external actuatable control valve, wherein operably coupled to the valve actuator compression mechanism, apparatus according to any one of claims 40 to 61.