CN102590087B - For the formation method of the sample that biological or chemical is analyzed - Google Patents

Abstract

One carries out biological or chemical method for measuring, described method comprises: (a) provides the fluidic hardware with sample area and the reacted constituent storage unit with multiple different reacted constituent for carrying out one or more mensuration, and described reacted constituent comprises sample generating component and sample analysis composition; B () makes sample generating component flow into according to predetermined scheme, to generate sample in described sample area; C () optionally controls the reaction conditions of described sample area, so that generate described sample; D () makes sample analysis composition flow into described sample area; E () detects the light signal sent from described sample area, described light signal indicates the dependent event between described sample analysis composition and described sample; Wherein (b)-(e) carries out in an automatic fashion.

Description

For the formation method of the sample that biological or chemical is analyzed

Technical field

Embodiments of the present invention relate generally to biological or chemical analytical approach and pick-up unit.

Background of invention

Various mensuration schemes for biological or chemical research relate to carries out a large amount of controlled reactions.In some cases, described controlled reaction carries out on a support surface.Then can observe and analyze described required reaction, to help attribute or the feature of determining chemicals involved in described required reaction.Such as, in some versions, under controlled conditions, the chemical composition part comprising discernible mark (such as fluorescence labeling) optionally can be bonded to another chemical composition part.By mark described in radiation excitation and detect from described mark light launch can observe these chemical reactions.Also can by other means (as chemiluminescence) provide as described in light launch.

The example of this kind of scheme comprises DNA sequencing.In synthesis limit, limit order-checking (SBS) scheme, on the surface of flow channel, form clonal expansion submanifold by bridge-type PCR.After the described clonal expansion submanifold of generation, can by described amplicon " linearization ", to produce single stranded DNA (sstDNA).A series of reagent is flowed into flow cell, to complete order-checking circulation.Each order-checking cycles through the fluorescently-labeled mononucleotide (such as A, T, G, C) with uniqueness and extends described sstDNA.Each nucleotide has the reversible terminator only allowing a single base of the interior generation of circulation to mix.After nucleotide is added to described sstDNA bunch, imaging in four passages (that is, each fluorescence labeling one).After imaging, described fluorescence labeling and described terminator are by from described sstDNA chemical cracking, and ever-increasing DNA chain prepares to be used for another circulation.Reagent conveying and the optical detection of several circulation can be repeated, to determine the sequence of described clonal expansion.

But the system being configured to perform these schemes may be limited in one's ability and may not meet cost benefit.Therefore, general system, the method and apparatus needing the improvement that can perform mensuration scheme (SBS scheme as described above) to have mode that is cost-benefit, that improve more easily or otherwise or can be used in described mensuration solution processes.

Summary of the invention

According to an embodiment, be provided for the fluidic hardware analyzing sample.Described fluidic hardware comprises the flow cell with entrance and exit and the flow channel extended betwixt.Described flow cell is configured to hold relevant sample.Described fluidic hardware also comprises shell, and described shell has the spatial accommodation being configured to hold described flow cell.Described spatial accommodation by allow described flow cell floating relative to described shell size making and shaping.Described fluidic hardware also comprises the packing ring being coupled to described shell.Described packing ring has entrance and exit passage and comprises compressible material.Described packing ring is arranged relative to described spatial accommodation, so as the described entrance and exit of described flow cell respectively with the described entrance and exit passage rough alignment of described packing ring.

In another embodiment, provide a kind of be configured to hold and be convenient to arrange dismountable chuck (cartridge) that flow cell carries out imaging.Described chuck comprises dismountable shell, and described dismountable shell has the spatial accommodation being configured to be remained on substantially by described flow cell in object plane.Described shell comprises a pair towards rightabout shell surface.Described spatial accommodation extends along at least one of described shell surface, so that described flow cell is exposed to the outside of described shell by least one of described shell surface.Described chuck also comprises the lid component that is coupled to described shell and comprises packing ring.Described packing ring has entrance and exit passage and comprises compressible material.Described gasket arrangements is for when described flow cell is by described outer casing supporting, and described packing ring is installed in the top of the expose portion of described flow cell.

In still another embodiment, provide a kind of setting for the method for the fluidic hardware of sample analysis.The stayed surface that described method is included in imaging system arranges dismountable fluidic hardware.Described device comprises spatial accommodation, is positioned at flow cell and the packing ring of described spatial accommodation.Described flow cell extends along the object plane in described spatial accommodation and can float relative to the described packing ring of described object plane inside.Described method to be also included in described spatial accommodation but on described stayed surface mobile described flow cell so that the entrance and exit passage rough alignment of the entrance and exit of described flow cell and described packing ring.

In another embodiment, provide a kind of setting for the method for the fluidic hardware of sample analysis.Described method comprises provides tool chlamydate fluidic hardware, and described shell comprises spatial accommodation and is positioned at relocatable flow cell of described spatial accommodation.Described shell has the groove closely adjacent with described spatial accommodation.Described method also comprises described fluidic hardware to be arranged on and has in the supporting construction of alignment members.Described alignment members is inserted into by corresponding groove.Described method also comprises the described flow cell in mobile described spatial accommodation.When described flow cell moves in described spatial accommodation, described alignment members engages the edge of described flow cell.

In another embodiment, provide a kind of fluidic hardware support, described fluidic hardware support is configured to relative to orthogonal X, Y and Z axis localizing sample region.Described device support comprises the supporting construction being configured to hold fluidic hardware.Described supporting construction comprise towards along described Z-direction basal plane and be configured to described device to be fixed thereon.Described device support also comprise multiple along XY plane separately direction reference field and comprise actuator and be operationally coupled to the alignment components of moveable registration arm of described actuator.Described registration arm has end of engagement.Described registration arm moves, to move described end of engagement towards with away from described reference field by described actuator between retraction and offset position.When described registration arm is in described offset position, described registration arm is configured to support described device against described reference field.

In another embodiment, provide a kind of fluidic hardware support, described fluidic hardware support comprises the supporting construction in the loading region had for holding fluidic hardware.Described supporting construction comprises the basal plane that partly limits described loading region and is configured to described device to arrange thereon.Described device support comprises the cap assemblies that is coupled to described supporting construction and is configured to be removably installed in the top of described device.Described cap assemblies comprises lid shell, and described lid shell has shell leg and the bridging part being connected described shell leg.Described shell leg extends with common direction and has therebetween viewing space.Described viewing is spatially located on described loading region.

In another embodiment, provide a kind of for the method relative to orthogonal X, Y and Z axis localizing sample region.Described method comprises provides alignment components, and described alignment components comprises the moveable registration arm with end of engagement.Described registration arm is moveable between retraction and offset position.Described method to be also included on the basal plane in the direction along described Z axis and to arrange fluidic hardware between multiple reference field towards the respective direction along XY plane.Described device has sample area.Described method also comprises mobile described registration arm to described offset position.Described device is pressed on described reference field by described registration arm, so that described device is maintained at fixed position.

In still another embodiment, provide a kind of optical module, described optical module comprises base plate, and described base plate has supporting surface and the parts spatial accommodation along described supporting surface.Described parts spatial accommodation is limited by reference field at least in part.Described optical module also comprises optics, and described optics has and is configured to reflected light or transmission light through optical surface wherein.Described optical module also comprises installing device, and described installing device has component holder and is operationally coupled to the biasing element of described retainer.Described retainer keeps described optics, so that the space segment of described optical surface is towards described reference field, the path sections of described optical surface extends beyond described supporting surface and enters light path.Described biasing element provides the aligning power of described optical surface against described reference field.In a particular embodiment, described parts spatial accommodation is the parts chamber extending into described base plate from the described supporting surface degree of depth of described base plate.Described optical surface and reference field can have the predetermined profile being configured to arrange by predetermined direction described optical surface.

In another embodiment, a kind of method of assembling optical system (opticaltrain) is provided.Described method comprises the base plate providing and have supporting surface and the parts spatial accommodation along described supporting surface.Described parts spatial accommodation is limited by reference field at least in part.Described method also comprises optics is inserted into described parts spatial accommodation.Described optics also comprises having and is configured to reflected light or transmission light through optical surface wherein.Described optical surface have towards described reference field space segment and extend beyond the path sections that described supporting surface enters light path.Described method also comprises to be provided the aligning power of described optical surface against described reference field.In a particular embodiment, described parts spatial accommodation is the parts chamber extending into described base plate from the described supporting surface degree of depth of described base plate.Described optical surface and reference field can have the predetermined profile being configured to arrange by predetermined direction described optical surface.

In another embodiment, provide a kind of optical imaging system, the subject support that described optical imaging system comprises fixing and mobile object and the detecting device detected from the light signal of the object of detector surface.Described imaging system also comprises and is configured to guide described light signal to the optical system in described detector surface.Described optical system has the object plane of closing on described subject support and the plane of delineation closing on described detector surface.Described optical system is included in rotatable mirror between image space and focal position.Described imaging system also comprises image analysis module, and when described mirror is in described focal position, described image analysis module is configured to analyze the test pattern detected in described detector surface.Described test pattern has best focal power (degree-of-focus) at the focal position of described test pattern.Indicate described object relative to the position of described object plane at the described focal position of described test pattern.Described subject support is configured to move described object based on described focal position towards described object plane.

In another embodiment, a kind of method of focus of control both optical imaging system is provided.Described method comprise provide be configured to guide light signal to the optical system in detector surface.Described optical system has the object plane of closing on object and the plane of delineation closing on described detector surface.Described optical system is included in rotatable mirror between image space and focal position.Described method also comprises the described mirror of rotation to described focal position, and when described mirror is at described focal position, obtains the test pattern of described object.Described test pattern has best focal power at the focal position of described test pattern.Described focal position indicates described object relative to the position of described object plane.Described method also comprises and moves described object based on described focal position towards described object plane.

In another embodiment, provide a kind of optical imaging system, described optical imaging system comprises the sample holder being configured to support flow cell.Described flow cell comprises the flow channel with sample area.Described imaging system also comprises and is coupled to described flow cell and is configured to guide reagent by described flow channel to the flow system of described sample area.Described imaging system also comprises and is configured to optical system on directs excitation light to described sample area and the first and second light sources.Described first and second light sources have the fixed position relative to described optical system.Described first and second light sources are provided for the first and second light signals exciting biomolecule respectively.Described imaging system also comprises the system controller being coupled to described first and second light sources and described flow system communicatedly.Described controller is configured to start described flow system to make described reagent flow into described sample area and to be configured to, after predetermined generated time section, activate described first and second light sources.Described light source can be, such as, and laser or semiconductor light sources (SLS) (as laser diode or light emitting diode (LED))

In another embodiment, a kind of method of carrying out biologicall test is provided.Described method comprises flow channel reagent being flow through have sample area.Described sample area comprises the biomolecule be configured to described reagent generation chemical reaction.Described method also comprises by sample area described in the first and second light source irradiation.Described first and second light sources provide the first and second light signals respectively.When being irradiated by described first or secondary light source, described biomolecule provides the light of instruction association reaction to launch.Described method also comprises the described light detected from described sample area and launches.Described light source can be, such as, and laser or semiconductor light sources (SLS) (as laser diode or light emitting diode (LED)).

In another embodiment, provide a kind of flow cell, described flow cell comprises and having towards reverse direction limit the mounting surface of a thickness and the ground floor of outside surface betwixt.Described flow cell also comprises and having towards reverse direction limit the channel surface of a thickness and the second layer of outside surface betwixt.The described second layer has the groove part extended along described channel surface.The described channel surface of the described second layer is fixed on described mounting surface.Described flow cell also comprises the flow channel limited by the described groove part of described channel surface and the planar section of described mounting surface.Described flow channel comprises imaging moiety.The described thickness of the described second layer is substantial uniform and be configured to transmitting optical signal through wherein along described imaging moiety.The described thickness of described ground floor is substantial uniform and be configured to allow uniform thermal power to be transmitted through wherein along described imaging moiety.

In another embodiment, provide a kind of light source module, described light source module comprises the module frame with optical channel and is fixed to described module frame and directed to guide light signal along the light source of light path by described optical channel.Described light source module also comprises and is fixed to described module frame and the optics of the fixed position had relative to described light source and predetermined direction.Described optics is placed in described optical channel, so that described optics is in described light path.

In another embodiment, provide a kind of exciting light module, described exciting light module comprises module frame and is fixed to first and second semiconductor light sources (SLS) of described module frame.Described first and second SLS have fixed position relative to each other.Described first and second SLS are configured to provide different optical excitation signals.Described exciting light module also comprises and is fixed to described module frame and has relative to the described fixed position of the first and second SLS and the optics of predetermined direction.Described optics allows the optical signal transmission from a described SLS pass wherein and reflect the light signal from described 2nd SLS.The light signal of described reflection and transmission is directed leaves described module frame along a common path.

In one embodiment, one is provided to carry out biological or chemical method for measuring.Described method is included between the fluidic hardware with sample area and the reacted constituent storage unit with multiple different reacted constituent for carrying out one or more mensuration and sets up fluid connection.Described reacted constituent comprises sample generating component and sample analysis composition.The described sample area that described method is also included in described fluidic hardware generates sample.Described generating run comprises makes different sample generating components flow into described sample area and control the reaction conditions of described sample area, to generate described sample.Described method also comprises the described sample analyzing described sample area.Described analysis operation comprises makes at least one sample analysis composition flow into described sample area.Described at least one sample analysis composition and described sample react, to provide the optically detectable signal of instruction dependent event.Described generation and analysis operation are carried out in an automatic fashion by described Analytical system.

In another embodiment, provide a kind of Analytical system, described Analytical system comprise be configured to supporting fluidic hardware also set up the fluidic hardware support be connected with the fluid of described fluidic hardware.Described Analytical system also comprises the fluidic networks being configured to described fluidic hardware fluid is connected to reacted constituent storage unit.Described Analytical system also comprises and is configured to optionally fluid be flowed into jet control system by described fluidic hardware from described storage unit.In addition, described Analytical system comprises the system controller with jet vectoring module.Described jet vectoring module be configured to jet control system (a) described in order with make different sample generating components from described storage unit flow into described sample area and the reaction conditions controlling described sample area to generate sample; And (b) flows into described sample area to make at least one sample analysis composition from described storage unit.Described at least one sample analysis composition is configured to react with described sample, to provide the optically detectable signal of instruction dependent event.Described Analytical system also comprises and is configured to detect the imaging system from optically detectable signal described in described sample.Described system controller is configured to automatically generate described sample by optionally controlling described fluidic hardware support, described jet control system and described imaging system and analyze described sample.

In another embodiment, one is provided to carry out biological or chemical method for measuring.Described method comprises: (a) provides the fluidic hardware with sample area and the reacted constituent storage unit with multiple different reacted constituent for carrying out one or more detections, and described reacted constituent comprises sample generating component and sample analysis composition; B () makes sample generating component flow into according to predetermined scheme, to generate sample in described sample area; C () optionally controls the reaction conditions of described sample area, so that generate described sample; D () makes sample analysis composition flow into described sample area; And (e) detects the light signal sent from described sample area, described light signal indicates the dependent event between described sample analysis composition and described sample; Wherein (b)-(e) carries out in an automatic fashion.

Accompanying drawing explanation

Fig. 1 is the block diagram of the Analytical system for carrying out biological or chemical mensuration according to an embodiment formation.

Fig. 2 is the side view being configured to the workstation carrying out biological or chemical mensuration according to an embodiment.

Fig. 3 is the front view of the described workstation of Fig. 2.

Fig. 4 is the figure of the fluidic networks according to an embodiment formation.

Fig. 5 is the skeleton view of the flow cell according to an embodiment formation.

Fig. 6 is the cross-sectional view of the described flow cell shown in Fig. 5 got along the line 6-6 in Fig. 5.

Fig. 7 is the planimetric map of the described flow cell of Fig. 5.

Fig. 8 is the enlarged drawing of the bending section of flow channel.

Fig. 9 is the skeleton view of the fluidic hardware according to an embodiment formation.

Figure 10 is another skeleton view of the described fluidic hardware of Fig. 9.

Figure 11 is the cross-sectional view of the described fluidic hardware of the Fig. 9 got along the line 11-11 in Fig. 9.

Figure 12 is the skeleton view of the fluidic hardware formed according to another embodiment.

Figure 13 is the skeleton view of the described fluidic hardware of Figure 12.

Figure 14 is the planimetric map of the fluidic hardware according to an embodiment formation.

Figure 15 is the side perspective view of the described fluidic hardware of Figure 14.

Figure 16 is the decomposed figure of the device support according to an embodiment formation.

Figure 17 is the skeleton view of the assembling support of Figure 16.

Figure 18 is the skeleton view of the supporting construction of the described support that can be used for Figure 16.

Figure 19 is the plan view from above of the described support of Figure 16.

Figure 20 be there is cap assemblies at aperture position Figure 16 described in the skeleton view of support.

Figure 21 is the amplification planimetric map of the described support of Figure 16.

Figure 22 is the skeleton view of the cap assemblies of the described support that can be used for Figure 16.

Figure 23 is the cross-sectional view of the described cap assemblies got along the line 23-23 shown in Figure 22.

Figure 24 is the skeleton view of the flow system that can use together with the described support of Figure 16.

Figure 25 is a kind of according to the block diagram of an embodiment setting for the method for the fluidic hardware of sample analysis.

Figure 26 illustrates a kind ofly to arrange block diagram for the method for the fluidic hardware of sample analysis according to embodiment.

Figure 27 be illustrate a kind of according to the block diagram of an embodiment for the method in localizing sample region.

Figure 28 is the skeleton view of the fluid storage system according to an embodiment formation.

Figure 29 is the cross-sectional side view of the described fluid storage system of Figure 28.

Figure 30 is the skeleton view pipetting assembly that can use together with the described fluid storage system of Figure 28.

Figure 31 is the skeleton view of the reacted constituent pallet according to an embodiment formation.

Figure 32 is the plan view from above of the described pallet shown in Figure 31.

Figure 33 is the side view of the described pallet shown in Figure 31.

Figure 34 is the front view of the described pallet shown in Figure 31.

Figure 35 is the cross-sectional side view in the composition hole that can use together with the described pallet of Figure 31.

Figure 36 is the bottom perspective view in the described composition hole of Figure 35.

Figure 37 is the skeleton view in the composition hole that can use together with the described pallet of Figure 31.

Figure 38 is the figure of the optical imaging system according to an embodiment.

Figure 39 is the skeleton view of the mobile control system according to an embodiment.

Figure 40 is the skeleton view of the parts that can use together with the described mobile control system of Figure 39.

Figure 41 is the skeleton view of the optical backplane of the described imaging system that can be used for Figure 38.

Figure 42 is the planimetric map of the described base plate of Figure 41.

Figure 43 is the skeleton view that can be used for the optics of the described imaging system of Figure 38 formed according to an embodiment.

Figure 44 is the broken-open perspective view of the described optics of Figure 43.

Figure 45 is the front view of the described optics of Figure 43.

Figure 46 is the side view of the described optics of Figure 43 in install operation process.

Figure 47 is the block diagram that a kind of method according to an embodiment assembling optical system is described.

Figure 48 is the skeleton view of the light source module according to an embodiment formation.

Figure 49 is the side view of the described light source module of Figure 48.

Figure 50 is the planimetric map of the described light source module of Figure 48.

Figure 51 is the planimetric map of the image focusing system according to an embodiment.

Figure 52 is the skeleton view of the rotatable mirror assembly of the described image focusing system that can be used for Figure 51.

Figure 53 is the schematic diagram being positioned at the rotatable mirror of image space of the described image focusing system that can be used for Figure 51.

Figure 54 and Figure 55 illustrates the sample image of the described image focusing system acquisition by Figure 51.

Figure 56 is the schematic diagram of the described rotatable mirror of the Figure 53 being positioned at focal position.

Figure 57 and 58 illustrates the test pattern of the described image focusing system acquisition by Figure 51.

Figure 59 is the block diagram of a kind of method that focus for control both optical imaging system is described.

Figure 60 illustrates that one is used for carrying out biological or chemical and analyzes method for measuring.

Figure 61 illustrates that one is used for carrying out biological or chemical and analyzes method for measuring.

Detailed Description Of The Invention

Embodiment as herein described comprises various system, method, assembly and device in order to detect reaction required in the sample analyzed for biological or chemical.In some embodiments, described required reaction provides the light signal detected by optical module.Described light signal can be launch from the light marked or can be the transmitted light by described sample reflection or refraction.Such as, embodiment can be used for performing or be convenient to perform the order-checking scheme that wherein sstDNA checks order in flow cell.In a particular embodiment, embodiment as herein described also can perform amplification scheme, to generate the associated sample for checking order.

As used herein, " required reaction " comprises stimulating the change of at least one making the chemistry of the material of response, electricity, physics and optical property or quality.Such as, described required reaction can be chemical transformation, chemical change or chemical action.In a particular embodiment, described required reaction is detected by imaging system.Described imaging system can comprise optical module light signal being guided to sensor (as CCD or CMOS).But in the mode that other are implemented, described imaging system can light signal described in direct-detection.Such as, flow cell can be installed on cmos sensor.But described required reaction also can be the change in electric property.Such as, described required reaction can be the change of ion concentration in solution.

Exemplary reaction includes but not limited to chemical reaction (as reduction, oxidation, addition, elimination, rearrangement, esterification, amidation, etherificate, cyclisation or replacement); Combination, wherein the first chemicals is bonded to the second chemicals; Dissociation reaction, wherein two or more chemicals is separated from each other; Fluorescence; Luminous; Chemiluminescence; And biological respinse (as nucleic acid replication, nucleic acid amplification, nucleic acid hybridization, nucleic acid connection, phosphorylation, enzymatic catalysis, receptors bind or ligand binding).Described required reaction also can be such as, be detected as addition or the elimination of the proton of the change of the pH value of surrounding medium or environment.

Described stimulation can be following at least one: physics, optics, electricity, magnetics with chemistry.Such as, described stimulation can be the exciting light of fluorophore in excited species.Described stimulation also can be the change of surrounding environment, as the concentration change of some biomolecule (as enzyme or ion) in solution.Described stimulation also can be the electric current being applied to solution in pre-qualified volume.In addition, described stimulation is by rocking, vibrating or reaction chamber residing for mobile described material provides, to produce power (such as centripetal force).As used herein, work done in the manner of a certain author extensive interpretation that phrase " makes response to stimulation " and comprise to stimulate make more directly reaction (such as, when after the incident exciting light of absorption, fluorophore sends the energy of specific wavelength) and because described stimulation startup finally causes the sequence of events of described response, more indirectly reaction is made (such as to stimulation, in Manganic pyrophosphate complex initiation, introduce alkali, finally cause chemiluminescence).Described stimulation can be (such as, incident on fluorophore exciting light) or progressive (such as, the temperature variation of surrounding environment) immediately.

As used herein, phrase " activity of the reaction needed for instruction " and variant thereof comprise available so that determine any event, attribute, quality or the feature detected whether required reaction occurs.The described activity detected can be the light signal generated in fluorescence or chemiluminescence.The described activity detected also can be that the electric property in pre-qualified volume or along the solution of predefined area changes.The described activity detected can be the change of temperature.

Various embodiment comprises provides reacted constituent to sample.As used herein, " reacted constituent " or " reactant " comprises the material of any reaction that can be used for needed for acquisition.Such as, reacted constituent comprises reagent, enzyme, sample, other biological molecule and buffer solution.Described reacted constituent is delivered to reaction site (region such as, residing for sample) in solution or usually at reaction site internal fixtion.Described reacted constituent directly or indirectly can react to relevant material.

In a particular embodiment, by reaction required described in optical module optical detection.Described optical module can comprise the optical system of the optics cooperatively interacted described light signal to be guided to image device (such as CCD, CMOS or photomultiplier).But, in substituting embodiment, the activity detector that described sample area can be set to be close to the described required reaction of detection and not use optical system.Described activity detector can have the ability to detect foregone conclusion part, attribute, quality or feature in pre-qualified volume or region.Such as, activity detector can have the ability to catch the image in described pre-qualified volume or region.In the solution that activity detector can have the ability to detect pre-qualified volume or along the ion concentration of predefined area.Exemplary activity detector comprises charge-coupled image sensor (CCD) (such as CCD camera); Photomultiplier (PMT); Characterization of molecules equipment or detecting device (as those use together with nano-pore as described in characterization of molecules equipment or detecting device); Microcircuit devices (full content of described patent is incorporated herein by reference for such as U.S. Patent number 7,595,883 those microcircuit devices described); And there is the CMOS sensor of field effect transistor (FET) (comprising chemical sensitive field effect transistor (chemFET), ion-sensitive field effect transistor (ISFET) and/or mos field effect transistor (MOSFET)).

As used herein, term " optics " comprises the various element affecting lightray propagation.Such as, described optics can have at least one function following: reboot, filtration, shaping, amplification or concentrate described light signal.May comprise from the light signal of described sample upstream and the light signal from described sample downstream by affected described light signal.In fluorescence detecting system, upstream components comprises those directs excitation radiation, and towards the parts of described sample, components downstream comprises the parts of those directs excitation radiation away from described sample.Optics can be, such as, and reverberator, dichroic mirror, beam splitter, collimating apparatus, lens, wave filter, chock, prism, mirror, detecting device etc.Optics also comprises bandpass filter, wedge and the optical device similar with those devices as herein described.

As used herein, term " light signal " comprises the electromagnetic energy that can be detected.The light that described term comprises from the biological or chemical material of mark is launched and also comprises the transmitted light being reflected by optical base-substrate or reflect.Light signal includes the exciting radiation be incident upon on described sample and the light provided by described sample is launched, and described light signal can have one or more spectral patterns.Such as, at imaging session, the mark of a more than type can be excited.In this case, dissimilar mark can by common excitation light source excites, or in the different time or at one time by different excitation light source excites.The mark of every type can send the light signal that its spectral pattern is different from other marks.Such as, described spectral pattern can have different emission spectrum.Described light is launched can be filtered, to detect the light signal from other emission spectrum respectively.

As used herein, term " different " launches (comprising emission spectrum or other emission characteristics) use for light, and described term broadly can be interpreted as comprising recognizable or differentiable light and launch.Such as, described photoemissive emission spectrum can have wavelength coverage overlapping at least partly, as long as a kind of emission spectrum is not exclusively overlapping with other emission spectrum at least partially.Different emission spectrum also can have same or similar wavelength coverage, but has different recognizable intensity.Different characteristic based on the exciting light producing described light signal can distinguish different light signals.Such as, in FRET (fluorescence resonance energy transfer) (FRET) imaging, described light is launched can be identical, but the described photoemissive origin cause of formation (such as optical excitation signal) may be different.More specifically, the first excitation wavelength can be used for the right donor fluorophore of excited donor-acceptor, so as FRET cause the transmitting from described acceptor and described acceptor excite the transmitting also will directly caused from described acceptor.On this point, distinguishing of described light signal based on to the observation transmitted, and can combine the confirmation to the described emission wavelength in order to produce described transmitting.Different light is launched can have other nonoverlapping features, as launched anisotropy or fluorescence lifetime.In addition, when described light is launched filtered, the wavelength coverage of described emission spectrum can be reduced.

Described optics can have fixing position in optical module, can be maybe optionally moveable.As used herein, term " optionally " uses together with " movement " and similar term, and described phrase refers to that the position of described optics can be changed in a desired manner.At least one of the position of described optics and direction can be changed.Such as, in a particular embodiment, rotatable mirror is optionally moved, so that focusing optical imaging system.

Different element as herein described and parts can be removably coupled.As used herein, when two or more elements or parts " are removably coupled " (or " removably installing " and other similar terms), described element can not destroyed the parts of described coupling by being easily separated.Such as, when element can easily separated from one another and without OVEREXERTION, do not use instrument (namely using hand) or do not spend a large amount of time in the separation of described parts, described element can be easily separable.For example, in some embodiments, optical device can be removably mounted to optical backplane.In addition, flow cell and fluidic hardware can be removably mounted to device support.

Imaging session comprises the time period be imaged at least partly of wherein said sample.A kind of sample can experience or stand multiple imaging session.Such as, a kind of sample can stand two different imaging session, and wherein each imaging session is attempted to detect the light signal from one or more not isolabelings.As a concrete example, the first at least part of scanning along nucleic acid samples can detect the mark relevant to nucleotide A and C and scan the mark that can detect and close with nucleotide G and T-phase along at least part of second of described sample.In order-checking embodiment, the independent stage can be there is in the alone cycle of order-checking scheme.Each circulation can comprise one or more imaging session.In other embodiments, the different sample of scanning can be comprised in different imaging session sensed light signal.Different samples can be identical type (such as two micro-array chips) or different types (such as flow cell and micro-array chip).

During imaging session, observe the light signal that described sample provides.Various types of imaging can use together with embodiment as herein described.Such as, embodiment as herein described can utilize " step-scan " method, and each several part coverlet of sample area alone becomes picture in the process.Embodiment also can be configured to execution and fall to penetrating at least one of fluorescence imaging and total internal reflection fluorescent (TIRF) imaging.In other embodiments, sample imager is postpone integrated (TDI) system sweep time.In addition, described imaging session can comprise " line scanning " one or more samples, so that the whole described sample of the line focus domain scanning of light.The description of some line scanning methods sees, such as, U.S. Patent number 7,329,860 and U.S. Patent Publication number 2009/0272914, wherein each entire subject matter is all incorporated to herein with way of reference.Imaging session also can comprise the point focusing district moving light with grating mode and cross over described sample.In substituting embodiment, imaging session can comprise when detecting unglazed photograph and the light transmitting of generation based on the emitting performance (radioactivity in such as described sample or chemiluminescence composition) of the mark in described sample completely.In substituting embodiment, flow cell can be installed on the imager (as CCD or CMOS) of the described required reaction of detection.

As used herein, term " sample " or " associated sample " comprise various relevant material or the material of experience imaging session, in described imaging session, observe the light signal from described material or material.In a particular embodiment, sample can comprise relevant biological or chemical material and alternatively, supports optical substrate or the supporting construction of described biological or chemical material.On this point, sample can comprise or can not comprise optical substrate or supporting construction.As used herein, term " biological or chemical material " can comprise the various biological or chemical material be suitable for optical system imaging as herein described or inspection.Such as, biological or chemical material comprises biomolecule, as nucleosides, nucleic acid, polynucleotide, oligonucleotides, protein, enzyme, polypeptide, antibody, antigen, part, acceptor, polysaccharide, carbohydrates, polyphosphate, nano-pore, organelle, lipid layer, cell, tissue, biosome and bioactive compound (analog or mimetic as mentioned kind).Other chemical substances comprise the mark that may be used for identifying, other marks that its example comprises fluorescence labeling and is hereafter described in further detail.

Dissimilar sample can comprise the different optical substrate or supporting construction that affect incident light in a different manner.In a particular embodiment, sample to be detected can be connected to one or more surfaces of substrate or supporting construction.Such as, flow cell can comprise one or more flow channel.In flow cell, described flow channel can be separated by the top layer of described flow cell and bottom and surrounding environment.Therefore, light signal projection to be detected also can be conveyed through multiple material layers with different refractivity from the inside of described supporting construction.Such as, when detecting from the light signal of the inner bottom surface of flow channel and when detection is from light signal above described flow channel, wish described light signal to be detected can propagate through there is a kind of refractive index fluid, there is one or more layer of different refractive indexes and the surrounding environment by having a kind of different refractivity by described flow cell.

As used herein, " fluidic hardware " a kind ofly comprises one or more flow channel of fluid that guides in a predetermined manner to carry out the device of required reaction.Described fluidic hardware is configured to be coupled to the fluidic networks of Analytical system by jet.For example, fluidic hardware can comprise flow cell or chip lab (lab-on-chip) equipment.Usually, flow cell along surface bearing sample with by outside imaging system images.Chip lab equipment can support sample and perform extra function, as utilized reaction required as described in integrated detector detection.Fluidic hardware can also comprise the parts that other are operably coupled to described flow channel alternatively, as shell or imager.In a particular embodiment, described passage can have the channel surface being provided with sample, and described fluidic hardware can comprise the transparent material allowing the imaging after required reaction occurs of described sample.

In a particular embodiment, described fluid device has the passage of microfluidic dimensions.In such passage, the bounding force flowed through between the surface tension of liquid wherein and the surface of cohesion and described liquid and described passage at least has materially affect to the flowing of described liquid.Such as, the cross-sectional area (intercepting perpendicular to flow direction) of microfluidic channel can be about 10 μm ²or it is less.

In substituting embodiment, optical imaging system as herein described can be used for scanning the sample with microarray.A microarray can comprise a different set of probe molecule being connected to one or more substrate, so that described different probe molecule can be distinguished from each other according to relative position.An array can comprise different probe molecules, or the probe molecule of different group, is wherein eachly positioned at addressed location different in substrate.Alternatively, a microarray can comprise independent optical substrate (as pearl), the probe molecule that each carrying one is different, or a different set of probe molecule, described probe molecule according to the position of described substrate on the surface being connected with described optical substrate or can identify according to described substrate position in a liquid.The exemplary array that wherein independent substrate is arranged on surface includes but not limited to, from inc. comprising the array of pearl in the BeadChip array of (SanDiego, CA) or other holes, is 6,266 as those are described in the patent No., 459,6,355,431,6,770,441,6,859,570 and 7, and the United States Patent (USP) of 622,294; And PCT publication No. is the array in the patent of WO00/63437, each of described patent is incorporated herein by reference.Other arrays from the teeth outwards with particle comprise those arrays of setting forth in US2005/0227252, WO05/033681 and WO04/024328 (wherein each be incorporated herein by reference).

Any one of various microarray known in the art can be used.Typical microarray comprises site (being also sometimes referred to as functional part), and each have one group of probe.Probe groups on each site has the probe of single variety usually, is homogeneity, but in some embodiments, each in described group can be heterogeneous.The site of array or functional part normally discrete, be spaced.Independent site can be continuous print, or they can have interval each other.Spacing between the size of described probe site and/or described site can be different, so that array can be high density, Midst density or comparatively low-density.The feature of high density arrays is that interval, site is less than about 15 μm.About 15 to 30 μm, the interval, site of Midst density array, and the interval, site of low-density array is greater than 30 μm.The site that interval is less than 100 μm, 50 μm, 10 μm, 5 μm, 1 μm or 0.5 μm can be had for array of the present invention.The resolution that the device of embodiment of the present invention or method can be used to be enough to the site distinguishing above-mentioned density or density range makes array image-forming.

The further example of operable commercially available microarray comprises, such as microarray or other are according to sometimes referred to as VLSIPS ^tMthe microarray of the method synthesis of (ultra-large immobilized polymer synthesis) technology, e.g., the such as patent No. is 5,324,633; 5,744,305; 5,451,683; 5,482,867; 5,491,074; 5,624,711; 5,795,716; 5,831,070; 5,856,101; 5,858,659; 5,874,219; 5,968,740; 5,974,164; 5,981,185; 5,981,956; 6,025,601; 6,033,860; 6,090,555; 6,136,269; 6,022,963; 6,083,697; 6,291,183; 6,309,831; 6,416,949; Described in the United States Patent (USP) of 6,428,752 and 6,482,591 (wherein each be incorporated herein by reference).Spotted microarrays also can be used in method according to the embodiment of the present invention.Exemplary spotted microarrays is the CodeLink from AmershamBiosciences ^tMarray.Another useful microarray is that a kind of ink jet printing method that uses is (from the SurePrint of AgilentTechnologies ^tMtechnology) microarray that manufactures.

The described system and method for setting forth herein can be used for detecting the existence of concrete target molecule in the sample of described microarray contact.This is passable, such as, determine based on the combination of target analyte to the concrete probe of described microarray of mark or the amendment of the target spot dependence due to concrete probe, to mix, to remove or to change the mark of described probe location.Any one in several detection can be used for utilizing microarray (e.g., such as publication No. be 2003/0108867,2003/0108900,2003/0170684, described in the U.S. Patent application of 2003/0207295 or 2005/0181394 (wherein each be incorporated herein by reference)) identify or characterize target spot.

In addition, optical system as herein described can be illustrated as the various parts described in the PCT application PCT/US07/07991 being entitled as " SystemandDevicesforSequencebySynthesisAnalysis " and assembly that comprise as submitted on March 30th, 2007 and/or comprise the international publication number being entitled as " FluorescenceExcitationandDetectionSystemandMethod " as submitted on September 26th, 2008 for the various parts described in the international application of WO2009/042862 and assembly the entire subject matter entirety of two applications (as described in be incorporated herein by reference).In a particular embodiment, optical system can comprise as U.S. Patent number 7, and 329,860 and the various parts described in WO2009/137435 (its entire subject matter entirety is incorporated herein by reference) and assembly.The optical system application number that also can comprise as submitted on Dec 15th, 2009 is the various parts described in United States Patent (USP) (its entire subject matter entirety is incorporated herein by reference) and the assembly of 12/638,770.

In a particular embodiment, method as herein described and optical system can be used for nucleic acid sequencing.Such as, order-checking (SBS) scheme in limit synthesis limit is particularly applicable.In SBS, multiple fluorescently-labeled modified ribonucleotide is used to upper multiple DNA amplification existed in surface (such as limiting the surface of passage in flow cell at least in part) to optical substrate bunch (may in 1,000,000 bunch) order-checking.Described flow cell can comprise the nucleic acid samples for checking order, and wherein said flow cell is placed in suitable flow cell support.The described sample for checking order can present the form of single nucleic acid molecule, described single nucleic acid molecule is separated from each other, so that as bunch or the nucleic acid molecules group that can split separately, increase of other characteristic formps, or be connected to the pearl of one or more nucleic acid molecules.Therefore, can check order on such as those arrays of setting forth above.Nucleic acid can be prepared as it and comprise Oligonucleolide primers in the position adjacent with unknown target sequence.Start a SBS order-checking circulation, the nucleotide of one or more not isolabeling and archaeal dna polymerase etc. flow into/flow through described flow cell by fluid flowing subsystem (not shown).The nucleotide of single type can once be added, or the nucleotide used in sequencing procedure can be specifically designed as has reversible terminated attribute, thus allow the sequencing reaction that each circulation occurs under the existence of the labeled nucleotide (as A, C, T, G) of several types simultaneously.Described nucleotide can comprise detectable mark part, as fluorophore.When described four kinds of nucleotide mix, described polymerase can select correct alkali to mix, and each sequence is extended by single base.Uncorporated nucleotide can be washed off by making washing lotion flow through described flow cell.One or more laser instruments can excite nucleic acid and bring out fluorescence.The fluorescence that described nucleic acid sends is based on the fluorophore of mixed alkali, and different fluorophores can send the utilizing emitted light of different wave length.Deblocking reagent can be added to described flow cell, to remove reversible terminator group from the DNA chain being extended and detecting.Then, described deblocking reagent can be washed off by making washing lotion flow through described flow cell.Then described flow cell is ready for use on the order-checking circulation of the introducing further starting from labeled nucleotide as above.Jet and detecting step can be repeated quickly and easily as many times as required, to complete sequencing procedures.The people such as the description of exemplary sequence measurement sees, such as Bentley, Nature456:53-59 (2008), WO04/018497; US7,057,026; WO91/06678; WO07/123744; US7,329,492; US7,211,414; US7,315,019; US7,405,281 and US2008/0108082, to be wherein eachly incorporated herein by reference.

In some embodiments, nucleic acid can order-checking before or period be attached to surface and increase.Such as, bridge amplification can be used to increase, to form nucleic acid bunch from the teeth outwards.The description of useful bridge amplification method sees, such as, U.S. Patent number 5,641,658, U.S. Patent Publication No. 2002/0055100, U.S. Patent number 7,115,400, U.S. Patent Publication No. 2004/0096853, U.S. Patent Publication No. 2004/0002090, U.S. Patent Publication No. 2007/0128624 and U.S. Patent Publication No. 2008/0009420.The method of another useful nucleic acid on surface of increasing is rolling circle amplification (RCA), such as, as people such as Lizardi, described in Nat.Genet.19:225-232 (1998) and US2007/0099208A1 (wherein each be incorporated herein by reference).On pearl, emulsion-based PCR also can use, such as, as people such as Dressman, described in Proc.Natl.Acad.Sci.USA100:8817-8822 (2003), WO05/010145 or U.S. Patent Publication No. 2005/0130173 or 2005/0064460 (wherein each full content is incorporated herein by reference).

Other sequencing technologies being applicable to the purposes of the described method and system of setting forth herein are Manganic pyrophosphate complex initiation, nano-pore order-checking and connection method order-checking.Useful especially exemplary pyrosequencing techniques and sample are described in US6,210, and 891, US6,258,568, US6,274,320 and Ronaghi, GenomeResearch11:3-11 (2001) (wherein each be incorporated herein by reference).Also useful exemplary nano hole technology and sample are described in the people such as Deamer, Acc.Chem.Res.35:817-825 (2002); The people such as Li, Nat.Mater.2:611-615 (2003); The people such as Soni, the people such as ClinChem.53:1996-2001 (2007), Healy, the people such as Nanomed.2:459-481 (2007) and Cockroft, J.am.Chem.Soc.130:818-820; And US7,001,792 (wherein each be incorporated herein by reference).Particularly, these methods utilize the agent delivery step repeated.Instrument in this paper or method may be configured with storage, valve, jet line and other fluidic component and the control system for those parts, to introduce reagent and according to required scheme (those schemes proposed in such as list of references as above) sensed light signal.Any one in various sample can be used to these systems, the substrate such as with the pearl generated by emulsion-based PCR, the substrate with zero mode waveguide, the substrate with integrated CMOS detecting device, the substrate in double-layer of lipoid with biological nano hole, the solid substrate with synthesis of nano hole and other substrates known in the art.These samples are described in various sequencing technologies backgrounds in list of references mentioned above and are described in US2005/0042648, US2005/0079510, US2005/0130173 and WO05/010145 (wherein each be incorporated herein by reference) further.

Can the exemplary indicia of detected according to different embodiments (such as, when being present in above supporting construction or time inner) include but not limited to chromophore, illuminophore, fluorophore, optical encoding nano particle, with the mark of the particle of diffraction grating coding, electrochemiluminescence (as Ru (bpy) ³²⁺) or the part that can be detected based on optical characteristics.Can comprise by useful fluorophore, such as fluoresce lanthanide compound (comprising the compound of those europiums and terbium), fluorescein, rhodamine, tetramethylrhodamine, eosin, erythrosine, cumarin, methylcoumarin, pyrene, malachite green, Cy3, Cy5, talan, fluorescein (LuciferYellow), CascadeBlue ^tM, glimmering and other fluorophores known in the art of TexasRed, alexa dyestuff, phycoerythrin, fluorine boron, as those are at Haugland, MolecularProbesHandbook, (Eugene, OR) 6thEdition; TheSynthegencatalog (Houston, TX.), Lakowicz, PrinciplesofFluorescenceSpectroscopy, the fluorophore described in 2ndEd., PlenumPressNewYork (1999) or WO98/59066 (wherein each be incorporated herein by reference).In some embodiments, a pair mark can be can be excited by the first excitation wavelength and another can be able to be excited by the second excitation wavelength mark.

Although embodiment is the example of the detection about the sample comprising the biological or chemical material supported by optical substrate, be appreciated that other samples can be imaged by embodiment as herein described.Other exemplary samples include but not limited to biological sample (as cell or tissue), electronic chip (as in computer processor use those) etc.The example of some application comprises microscope, satellite scanner, high resolving power duplicating, fluoroscopic image collection, foranalysis of nucleic acids and order-checking, DNA sequencing, the order-checking of synthesis limit, limit, Microarray image, the imaging of holographic encoding particulate etc.

Fig. 1 is the block diagram of the Analytical system 100 for carrying out biological or chemical analysis according to an embodiment formation.In some embodiments, described Analytical system 100 is the workstations that can be similar to bench device or desktop computer.Such as, at least great majority can be co-located in the shell 117 of described Analytical system 100 for the system and parts of carrying out required reaction.In other embodiments, described Analytical system 100 comprises from one or more parts of the long-range setting of described Analytical system 100, assembly or system (such as remote data base).Described Analytical system 100 can comprise interact with each other with the various parts performing one or more preordering methods for biological or chemical analysis or the scheme of mensuration, assembly and system (or subsystem).

Such as, described Analytical system 100 comprises system controller 102, and described system controller 102 can be communicated with system (or subsystem) with the described various parts of described Analytical system 100, assembly.As shown in the figure, described Analytical system 100 has optical module 104, excitaton source assembly 106, detector module 108 and supports one or more fluidic hardware support 110 it with the fluidic hardware 112 of sample.Described fluidic hardware can be flow cell, flow cell 200 as mentioned below, or described fluidic hardware 112 can be fluidic hardware 300 hereinafter described.

In some embodiments, described optical module 104 is configured to the incident light of guiding from described excitaton source assembly 106 on described fluidic hardware 112.Described excitaton source assembly 106 can comprise the excitation source that one or more are configured to excite the mark relevant to described sample.Described excitaton source assembly 106 also can be configured to the incident light provided by described sample reflection and/or refraction.As shown in the figure, described sample can provide and comprise the light signal that light launches 116 and/or transmitted light 118.Described device support 110 and described optical module 104 can move relative to each other.In some embodiments, described device support 110 comprises the electric machine assembly 132 moving described fluidic hardware 112 relative to described optical module 104.In other embodiments, described optical module 104 can be moved to described device support 110 in addition or alternatively.Described optical module 104 also can be configured to guide described light transmitting 116 and/or transmitted light 118 to described detector module 108.Described detector module 108 can comprise one or more imaging detectors.Described imaging detector can be, just as an example, and CCD or CMOS camera, or photomultiplier.

Still as shown in the figure, described Analytical system 100 can comprise the jet control system 134 of the fluid flowing controlling whole fluidic networks 135 (solid line represents).Described jet control system 134 can be in, such as, sends reacted constituent (such as reagent) or other fluids during order-checking scheme to described fluidic hardware 112.Described Analytical system 100 also can comprise be configured to deposit described Analytical system 100 can fluid fluid storage system 136 and regulate the temperature control system 138 of temperature of described fluid.Described temperature control system 138 usually also can utilize, and such as radiating module, heating radiator and fan blower regulate the temperature of described Analytical system 100.

Still as shown in the figure, described Analytical system 100 can comprise the user interface 140 with user interaction.Such as, described user interface 140 can comprise display or ask the display 142 from the information of user and the user input device 144 receiving user's input.In some embodiments, described display 142 and described user input device 144 are identical equipment (as touch-screen).As will be hereafter described in a more detailed discussion, described Analytical system 100 can be communicated with all parts to perform required reaction.Described Analytical system 100 also can be configured to analysis detecting data, to provide information needed to user.

Described jet control system 134 is configured to guide and regulate one or more fluids by described fluidic networks 135.Described jet control system 134 can comprise, and such as, optionally can be used to the pump and valve that control fluid flowing.Described fluidic networks 135 can be communicated with described fluid storage system 136 fluid with described fluidic hardware 112.Such as, selected fluid can be drawn from described fluid storage system 136 and is directed to described fluidic hardware 112 in a controlled manner, or described fluid can be drawn from described fluidic hardware 112 and is drawn towards, such as, litter receptacle in described fluid storage system 136.Though do not show, described jet control system 134 also can comprise the flow velocity of described fluid or the flow sensor of pressure in the described fluidic networks of detection.Described sensor can be communicated with described system controller 102.

Described temperature control system 138 is configured to the fluid temperature (F.T.) of the zones of different regulating described fluidic networks 135, described fluid storage system 136 and/or described fluidic hardware 112.Such as, described temperature control system 138 can comprise and to engage with described fluidic hardware 112 and to control the thermo cycler 113 of the temperature of the fluid flowed along described fluidic hardware 112.Though do not show, described temperature control system 138 can comprise the sensor of the temperature of test fluid or miscellaneous part.Described sensor can be communicated with described system controller 102.

Described fluid storage system 136 is communicated with described fluidic hardware 112 fluid and can stores the various reacted constituent or reactant that are used for carrying out required reaction herein.Described fluid storage system 136 can store for cleaning or clean described fluidic networks 135 or described fluidic hardware 112 and also for diluting the fluid of described reactant.Such as, described fluid storage system 136 can comprise the various storage for storing reagent, enzyme, other biological molecule, buffer solution, water and non-polar solution etc.In addition, described fluid storage system 136 also can comprise the litter receptacle for receiving waste product.

Described device support 110 is configured to such as, with mechanical, electrically and in fluid application the one or more described fluidic hardware 112 of at least one engagement.Described device support 110 can support described fluidic hardware 112, so that fluid flows through described fluidic hardware 112 and/or described fluidic hardware 112 imaging in a direction expected.

Described system controller 102 can comprise any based on processor or the system based on microprocessor, comprise use microcontroller, reduced instruction set computers (RISC), special IC (ASIC), field programmable gate array (FPGA), logical circuit and any other can perform the circuit of function as herein described or the system of processor.Example is above only exemplary, and is therefore not necessarily intended to the definition and/or the implication that limit described term system controller.In an exemplary embodiment, described system controller 102 performs the instruction group being stored in one or more storage unit, storer or module, to realize detecting the acquisition of data and at least one of analysis.Memory element can be the form of the physical memory element in information source or described Analytical system 100.

Described instruction group can comprise the order that the described Analytical system of various instruction 100 performs concrete operations (Method and Process of various embodiment as described herein).Described instruction group can be the form of software program.As used herein, term " software " and " firmware " can exchange, and comprising any computer program that storage performs by computing machine in memory, described storer comprises RAM storer, ROM storer, eprom memory, eeprom memory, non-volatile ram (NVRAM) storer.Above type of memory is only exemplary, because of instead of to the restriction of type that can be used for the storer storing computer program.

Described software can be various forms, as system software or application software.In addition, described software can be the set of independent program, or compared with the program module of large program inside or a part for program module.Described software also can comprise the modularization programming of the form of object based programming.Obtain and detect after data, automatically process, respond user's input processing or respond the request (such as by the remote request of communication link) that another processor makes by described Analytical system 100 and process described detection data.

Described system controller 102 is connected to miscellaneous part or the subsystem of described Analytical system 100 by communication link (dotted line represents).Described system controller 102 also can be communicably connected to remote system or server.Described communication link can be hardwired or wireless.The user that described system controller 102 can receive from described user interface 140 inputs or order.Described user input device 144 can comprise keyboard, mouse, touch panel and/or speech recognition system etc.Alternatively or additionally, described user input device 144 also can be described display 142.

Fig. 1 also illustrates the block diagram of described system controller 102.In one embodiment, described system controller 102 comprises one or more processor that can communicate with each other or module.Described system controller 102 is conceptually illustrated by the set as module, but the combination in any of specialized hardware plate, DSP, processor etc. can be utilized to realize.Alternatively, described system controller 102 can utilize the ready-made PC with uniprocessor or multiprocessor to realize, and feature operation is distributed between described processor.As a further alternative, module hereinafter described can utilize mixed configuration to realize, and wherein certain module function utilizes specialized hardware to perform, and remaining functions of modules utilizes ready-made PC etc. to perform.The software module that described module also can be used as in processing unit realizes.

Described system controller 102 can comprise the multiple module 151-158 be communicated with system control module 150.Described system control module 150 can be communicated with described user interface 140.Directly be communicated with described system control module 150 although described module 151-158 is shown, described module 151-158 also can each other, with described user interface 140 or be directly communicated with other system.In addition, described module 151-158 can be communicated with described system control module 150 by other modules.

Described multiple module 151-158 comprises the system module 151-153 be communicated with described subsystem.Described fluid control module 151 can be communicated with described jet control system 134, to control valve and the flow sensor of described fluidic networks 135, thus controls one or more fluids and flows through described fluidic networks 135.When fluid step-down or when must change when described litter receptacle, described fluid storage module 152 can notify user.Described fluid storage module 152 can also be communicated with described temperature control modules 153, so that described fluid can be stored at desirable temperature.

Described multiple module 151-158 also can comprise and receives and analyze the image analysis module 158 of the detection data (such as view data) from described detector module 108.Detection data after process can be stored and maybe can be sent to described user interface 140 to show required information to user for subsequent analysis.Scheme module 155-157 is communicated with described system control module 150, to control the running of described subsystem when carrying out predetermined mensuration scheme.Described scheme module 155-157 can comprise and is used to indicate described Analytical system 100 and performs the instruction group of concrete operations according to predetermined scheme.

Described scheme module 155 can be configured to be emitted in the order generating sample in described fluidic hardware 112.Such as, described scheme module 155 can indicate described fluid storage system 136 and described temperature control system 138 to generate sample in sample area.In a concrete embodiment, described scheme module 155 can send the order performing bridge-type PCR, and wherein clonal expansion submanifold is formed at the partial zones in flow cell passage (or path).

Described scheme module 156 can be synthesis limit, limit order-checking (SBS) module being configured to the order sending synthesis limit, various execution limit sequencing procedure.In some embodiments, described SBS module 156 also can process detection data.After generating described amplicon by bridge-type PCR, described SBS module 156 can provide the instruction carrying out described amplicon linearization or sex change to produce sstDNA and to add sequencing primer, so that described sequencing primer can hybridize the universal sequence becoming and be positioned at domain of dependence side.Each order-checking is cycled through sstDNA described in Single base extension and is completed by modification archaeal dna polymerase and four kinds of mixture of ribonucleotides (it is sent and can be subject to the instruction of described SBS module 156).Dissimilar nucleotide has unique fluorescence labeling, and each nucleotide has the reversible terminator only allowing a single base of the interior generation of circulation to mix.After single base is added into described sstDNA, described SBS module 156 can send the instruction of cleaning step, to remove uncorporated nucleotide by making washing lotion flow through described flow cell.Described SBS module 156 can indicate described excitaton source assembly and detector module to perform imaging session to detect fluorescence (i.e. each fluorescence labeling one) in one of described four passages further.After imaging, described SBS module 156 can indicate sends deblocking reagent, with from fluorescence labeling and described terminator described in described sstDNA chemical cleavage.Described SBS module 156 can send the instruction of cleaning step, to remove the product of described deblocking reagent and described deblocking reagent.Can carry out subsequently another kind of like order-checking circulation.In this order-checking scheme, described SBS module 156 can indicate described jet control system 134 to guide reagent and enzyme solutions to flow through described fluidic hardware 112.

In some embodiments, described SBS module 157 can be configured to the order sending various execution Manganic pyrophosphate complex initiation protocol step.When concrete nucleotide is incorporated into nascent strand, Manganic pyrophosphate complex initiation detects the release (Ronaghi of inorganic pyrophosphate (PPi), M. people is waited. (1996) " Real-timeDNAsequencingusingdetectionofpyrophosphaterelea se. " AnalyticalBiochemistry242 (1), 84-9; Ronaghi, M. (2001) " PyrosequencingshedslightonDNAsequencing. " GenomeRes.11 (1), 3-11; The people such as Ronaghi, M.. (1998) " Asequencingmethodbasedonreal-timepyrophosphate. " Science281 (5375), 363; U.S. Patent number 6,210,891; U.S. Patent number 6,258,568 and U.S. Patent number 6,274,320, its full content disclosed is incorporated herein by reference).In Manganic pyrophosphate complex initiation, the PPi of release can detect by being converted into ATP immediately by adenosine triphosphate adenosine monophosphate (ATP) sulfurylase, and the photon that the ATP generated produces horizontally through luciferase detects.In this case, described fluidic hardware 112 can comprise millions of holes, and wherein each hole has the single trapping pearl with clonal expansion sstDNA thereon.Each hole also can comprise other less pearls (such as can carry immobilised enzymes (such as ATP sulfurylase and luciferase) or be convenient to catch pearl maintenance pearl in the hole by described).Described SBS module 157 can be configured to give an order to described fluid control module 151, to carry out circulation (such as, the 1st circulation: the A of the fluid carrying single type nucleotide continuously; 2nd circulation: G; 3rd circulation: C; 4th circulation: T; 5th circulation: A; 6th circulation: G; 7th circulation: C; 8th circulation: T; Etc.).When nucleotide is incorporated in DNA, pyrophosphoric acid is released, thus causes chain reaction, wherein produces the burst of light.The described burst of light can be detected by the sample detector of described detector module.Detection data can be communicated to described system control module 150, described image analysis module 158 and/or described SBS module 157 and process.Described detection data can store for later analysis or be undertaken analyzing by described system controller 102 and image can be sent to described user interface 140.

In some embodiments, user can provide user to input by described user interface 140, to select the mensuration scheme will run by described Analytical system 100.In other embodiments, described Analytical system 100 automatically can detect the type of the fluidic hardware 112 being inserted into described device support 110 and confirm mensuration scheme to be run with user.Alternatively, described Analytical system 100 can provide a limited number of mensuration scheme run with the fluidic hardware 112 determining type.User can select required mensuration scheme, then described Analytical system 100 can according to preprogrammed instruction perform the mensuration scheme selected.

Fig. 2 and 3 illustrates the workstation 160 that the biological and chemical being arranged to sample according to an embodiment is analyzed.As shown in the figure, described workstation 160 is relative to orthogonal X, Y and Z axis location.In the embodiment shown, terrestrial gravitation g is parallel to the extension of described Z axis.Described workstation 160 can comprise workstation shell 162 (or workstation shell), and it is shown in the cut-open view of Fig. 2 and Fig. 3.Described shell 162 is configured to the various elements holding described workstation 160.Such as, described workstation 160 can comprise the like about described Analytical system 100 (Fig. 1) as described above.As shown in the figure, described workstation 160 has optical table 164, and described optical table 164 has multiple optics being mounted to it.Described optics can optical module (as according to Figure 38 wait describe as described in optical module 602) a part.Described optical table 164 can have the fixed position relative to described shell 162.

Described workstation 160 also can comprise the sample stage 166 being coupled to described optical table 164 movably.Described sample stage 166 can have the sliding platform 168 supporting and have the fluidic hardware of relevant sample thereon.In the embodiment shown, described fluidic hardware is the fluidic hardware 300 hereafter described in more detail.Described platform 168 be configured to relative to described optical table 164 and, more specifically, slide relative to the imaging len of described optical module 602.For this reason, described platform 168 can along described X-axis Bidirectional slide, so that described fluidic hardware 300 can be placed on described sample stage 166 and so that described imaging len can slide the sample imaging made wherein above described fluidic hardware 300.In other embodiments, described platform 168 can be that static and described sample stage 166 can along described X-axis Bidirectional slide, to arrange described fluidic hardware 300 relative to the imaging len of described optical module 602.Therefore, described platform and sample stage can be relative to each other moveable due to described sample stage, platform or both movements.

Still as shown in the figure, described workstation 160 can comprise user interface 172, computing system 174 (Fig. 2) and fluid storage unit 176 and 178 (Fig. 4).Described user interface 172 can be configured to show information to user and also receive the touch-screen of user's input.Such as, described touch-screen can receive the order performing predetermined mensuration scheme or receive from the inquiry of user.Described computing system 174 can comprise processor and module, as according to Fig. 1 describe as described in system controller 102 and as described in module 151-158.Described fluid storage unit 176 and 178 can be a part for larger fluid storage system.Described fluid storage unit 176 can be used for the refuse of the described mensuration generation of collection execution and described fluid storage unit 178 can comprise damping fluid.

Fig. 4 is the figure of the fluidic networks 552 that can be used for described workstation 160 (Fig. 2).As used herein, fluid can be liquid, gel, gas or its potpourri.In addition, fluid can be the potpourri of two or more liquid.Described fluidic networks 552 can comprise multiple fluidic component (such as, fluid line, pump, flow cell or other fluidic hardwares, manifold, storage) being configured to one or more fluids and flowing through.As shown in the figure, described fluidic networks 552 comprises multiple fluidic component 553-561 (solid line represents) interconnected by fluid line.In the embodiment shown, described fluidic networks 552 comprises buffer solution container 553, reagent tray 554, multi-ported valve 555, bypass valve 556, flow sensor 557, flow cell 558, another flow sensor 559, pump 560 and litter receptacle 561.The flow direction of fluid is by representing along the arrow of described fluid line.Except described fluidic component 553-561, described fluidic networks also can comprise other fluidic component.

Described reagent tray 554 can be similar with the reacted constituent pallet hereafter described in more detail (or reacted constituent storage unit) 1020.Described pallet 1020 can comprise the various container (such as bottle or pipe) comprised for performing the reacted constituent measured with embodiment as herein described.The operation of described multi-ported valve 555 can be controlled by Analytical system, as described in Analytical system 100, flow into described flow cell 558 optionally to make different fluids (comprising its potpourri).Described flow cell 558 can be described flow cell 200 or described fluidic hardware 300 (both are hereafter being described in more detail) or other suitable fluidic hardwares.

Fig. 5-60 (being hereafter described in more detail) illustrates the various elements (such as parts, equipment, assembly, system etc.) and method that can use together with described workstation 160.These elements can be fitted to each other with Imaged samples, analyze described detection data and user to described workstation 160 provides information.But element below and method also can independently use, in other devices use or use together with other devices.Such as, described flow cell 200 and described fluidic hardware 300 can be used for other Analytical system.Described optical module 602 (and element) can be used for checking sundry item, as microcircuit.In addition, described device support 400 can be used to support other fluidic hardwares, as chip lab equipment.Analytical system with these equipment can comprise or not comprise optical module, to detect described required reaction.

Fig. 5-7 illustrates the flow cell 200 formed according to an embodiment.As illustrated in figs. 5-7, described flow cell 200 is relative to described X, Y and Z axis location.Described flow cell 200 is configured to keep relevant sample 205 in flow channel 206.Described sample 205 is shown in SBS solution processes can multiple DNA bunch of imaging, but other samples can be used in substituting embodiment.Although only illustrate single U-shaped flow channel 206, substituting embodiment can comprise the flow cell of the many flow channels had with difform path.Described flow cell 200 can be communicated with fluidic system (not shown) fluid, and described fluidic system is configured to the described sample 205 in delivery of agents to described flow channel 206.In some embodiments, after required reaction occurs, described sample 205 can provide detectable feature (such as by fluorescence or chemiluminescence).Such as, described flow cell 200 can have light signal from the one or more sample area wherein sent or part (region namely residing for described sample 205 or part).In some embodiments, described flow cell 200 also can be used for generating described sample 205, measures to perform biological or chemical.Such as, before described SBS scheme performs, described flow cell 200 can be used for generating described DNA bunch.

As illustrated in figs. 5-7, described flow cell 200 can comprise and is fixed together and limits ground floor 202 and the second layer 204 of described flow channel 206 betwixt.Described ground floor 202 has mounting surface 208 and outer surface or outside surface 210 (Fig. 5 and 6).Described mounting surface and outer surface 208 and 210 be in the opposite direction and betwixt thickness limited T along described Z axis face ₁(Fig. 5 and 6).Described thickness T ₁uniform substantially along XY plane, but can be different in substituting embodiment.The described second layer 204 has channel surface 212 (Fig. 6) and outer surface or outside surface 214.Described channel surface and outer surface 212 and 214 be in the opposite direction and betwixt thickness limited T along described Z axis face ₂(Fig. 6).

Still as shown in Figure 5, described ground floor 202 has the size or length L measured along described X-axis ₁with another size measured along described Y-axis or width W ₁.In some embodiments, described flow cell 200 is named as microdevice.Microdevice may be difficult to be kept by the hand of individual or move.Such as, the described length L of described flow cell 200 ₁can be about 100mm or about 50mm or less.In a particular embodiment, described length L ₁about 30mm or less.In some embodiments, described width W ₁can be about 35mm or about 25mm or less, more specifically, described width W ₁can be about 15mm or less.In addition, the merging shown in Fig. 7 or overall height H _t(such as thickness T ₁and T ₂summation) can be about 10mm or about 5mm or less.More specifically, described height H _tcan be about 2mm or about 1.5mm or less.

Described flow cell 200 comprises edge 231-234 linearly in the embodiment shown.Edge 231 and 233 is with described width W ₁spaced apart and extend the described length L of described flow cell 200 ₁.Edge 232 and 234 is with described length L ₁spaced apart and along described width W ₁extend.Still as shown in the figure, the described second layer 204 can have the size or length L measured along described X-axis ₂with another size measured along described Y-axis or width W ₂.In the embodiment shown, described edge 231-234 limits the periphery of described flow cell 200 and extends along the common pond plane being parallel to the extension of described XY plane.Still as shown in the figure, the described second layer 204 can have as described in the edge 241-244 (as shown in Figure 5) of the similar location of edge 231-234.

In the embodiment shown, described width W ₁be greater than described width W substantially ₂, and the described second layer 204 is only placed in a part for described mounting surface 208.On this point, described mounting surface 208 comprises retained part 208A and 208B of exposure in the relative both sides of the described second layer 204.Described width W ₂extend between described retained part 208A and 208B.Described flow cell 200 also can have along described Z axis towards face, rightabout pond 256 and 258.In the embodiment shown, face, described pond 256 comprises described retained part 208A and 208B and described outside surface 214, and face, described pond 258 comprises described outside surface 210.Still as shown in the figure, described flow cell 200 can be extending longitudinally between the first and second relative pond ends 246 and 248.In the embodiment shown, on described first pond end 246, coplanar and described edge 234 is coplanar each other substantially on described the second relative pond end 248 with 244 each other substantially with 242 at described edge 232.

As shown in Figure 6, the described second layer 204 has the groove part 216 that at least one extends along described channel surface 212.In the embodiment shown, described channel surface 212 is etched with forms described trench portions 216, but described trench portions 216 can pass through other techniques (as channel surface 212 as described in cutting) formation.For assembling described flow cell 200, the described channel surface 212 of the described second layer 204 is mounted and is fixed in the described mounting surface 208 of described ground floor 202.Such as, described channel surface and mounting surface 212 and 208 can use and prevent the bonding agent of the leakage from described flow cell 200 (such as photoactivation resin) to be bonded together.In other embodiments, described channel surface and mounting surface 212 and 208 are bonded together by other bonding agents or mechanical interlocked and/or be fixed together.Therefore, described ground floor 202 is configured to cover the described groove part 216 of the described second layer 204 to form described flow channel 206.In the embodiment shown, described trench portions 216 can be extend described length L substantially towards described first end ₂, bending, then extend described length L substantially towards described second end ₂single continuous print groove.Therefore, described flow channel 206 can be U-shaped substantially.

Fig. 5-7 shows described sample 205 and only arranges along described mounting surface 208.But in other embodiments, described sample 205 can be placed on the surface of the described flow channel 206 of any restriction.For example, described sample 205 also can be placed on the matching surface 212 of the described trench portions 216 partly limiting described flow channel 206.

In the embodiment shown, described flow channel 206 can comprise multiple channel section 250-252.Different channel sections can have the different size relative to adjacent upstream or downstream passage section.In the embodiment shown, described flow channel 206 can comprise channel section 250 (it also can be called as imaging session 250).Described channel section 250 can have the sample area be configured to by the imaging of imaging system (not shown).Described flow channel 206 also can have channel section 251 and 252 (it also can be called as non-imaged section 251 and 252).As shown in the figure, described channel section 250 and 252 extends parallel to each other by described flow cell 200.The described channel section 251 and 252 of described flow channel 206 can be pressed certain size relative to described channel section 250 and to make and shaping, to control to flow through the flowing of fluid wherein and gas.

Such as, Fig. 7 also illustrates described channel section 250-252 xsect C separately ₁-C ₃, it passes through perpendicular to flow direction F ₁intercept.In some embodiments, described xsect C ₁-C ₃can be different size (namely different sectional areas), to control the flowing of fluid by described flow channel 206.Such as, described xsect C ₁size be greater than described xsect C ₂and C ₃.More specifically, the described channel section 250-252 of described flow channel 206 can have height H equal substantially ₁, described height H ₁measure between the described groove part 216 (Fig. 6) and described mounting surface 208 of described channel surface 212.But the described channel section 250-252 of described flow channel 206 can have different width W respectively ₃-W ₅.Described width W ₃be greater than described width W ₄and W ₅.Described channel section 251 connects the bending of described channel section 250 and 252 or bend loss with can forming jet.Described xsect C ₃than described xsect C ₁and C ₂less.Such as, described width W ₅be less than described width W ₃and W ₄.

Fig. 8 is the enlarged drawing of the part of described bending section 251 and described channel section 250 and 252.As mentioned above, described channel section 250 and 252 can extend parallel to each other.In described flow channel 206, perhaps need the Uniform Flow by described sample area.Such as, described fluid can comprise stream part F ₂-F ₄.The size of described channel section 250-252 can be configured, so that described stream part F ₂-F ₄in whole described sample area, there is flow velocity equal substantially.In this kind of embodiment, different section or the part (Fig. 5) of described sample 205 can have the time of equivalent substantially to react with the reacted constituent in described fluid.

For this reason, the described bending section 251 of described flow channel 206 connects discrete profile of described channel section 250 and 252 with can having jet.Such as, as shown in Figure 8, described bending section 251 can comprise conical section 270, center section 276 and downstream part 278.As shown in the figure, described conical section 270 has the width W that size reduces gradually _5A.More specifically, described bending section 251 can comprise the sidewall 272 and 274 extended internally toward each other with angle equal substantially.Described center section 276 bends from described conical section 270 to described downstream part 278.The width W that the size that has described center section 276 reduces gradually, then size starts increase _5B.Described downstream part 278 has substantial uniform width W everywhere _5Cand extend to described channel section 252 along path linear substantially from described center section 276.In other words, described sidewall 272 and 274 can extend parallel to each other through described downstream part 278.

Turn back to Fig. 7, described flow cell 200 includes an inlet and an outlet, and is respectively 222 and 224.Described entrance and exit 222 and 224 is only had and is formed by the described second layer 204.But in substituting embodiment, described entrance and exit 222 and 224 only has by described ground floor 202 or by described both layers 202 and 204 formation.Described flow channel 206 and described entrance and exit 222 are communicated with 224 fluids and extend wherein.In a particular embodiment, described entrance and exit 222 and 224 abuts one another (or adjoining with described edge 234 and 244) at the described pond end 248 of described flow cell 200.Such as, the interval 282 being separated described entrance and exit 222 and 224 can approximate described width W ₃.More specifically, described interval 282 can be about 3mm, about 2mm about or less.In addition, described channel section 250 and 252 can be separated by interval 280.Described interval 280 can be less than the described width W of described channel section 250 ₃or more specifically, be less than the described width W of described channel section 252 ₄.Therefore, the path of described flow channel 206 can be substantially U-shaped and, in the embodiment shown, there is the discrete profile along described bending section 251.

In substituting embodiment, described flow channel 206 can have different paths, so that described entrance and exit 222 and 224 has different positions in described flow cell 200.Such as, described flow channel can form the unipath of the described outlet extending to described flow cell opposite end from the described entrance of described flow cell one end.

About Fig. 6, in some embodiments, the described thickness T of the described second layer 204 ₂(Fig. 6) be substantial uniform along described imaging moiety 250.Along the described uniform thickness T of described imaging moiety 250 ₂transmitting optical signal can be configured to pass wherein.In addition, the described thickness T of described ground floor 202 ₁be substantial uniform along described imaging moiety 250 and be configured to be transferred through and wherein enter described flow channel 206 with allowing uniform thermal power.

Fig. 9-11 illustrates the fluidic hardware 300 formed according to an embodiment.In order to illustrative object, described fluidic hardware 300 is relative to orthogonal X, the Y shown in Fig. 9 and 10 and Z axis location.Fig. 9 and 10 is skeleton views of described fluidic hardware 300.As shown in Figures 9 and 10, described fluidic hardware 300 comprises chuck (or flow cell carrier) 302 and described flow cell 200.Described chuck 302 is configured to hold described flow cell 200 and is convenient to locate the described flow cell 200 for imaging session.

In some embodiments, described fluidic hardware 300 and described chuck 302 can be dismountable, so that described chuck 302 can be removed from imaging system (not shown) by individual or machine and not damaged described fluidic hardware 300 or chuck 302.Such as, described chuck 302 can be configured to repeatedly be inserted and moves to described imaging system and do not destroy described chuck 302 or make described chuck 302 be not suitable for its expection object.In some embodiments, described fluidic hardware 300 and described chuck 302 can make by certain size and shaping with operated by individual.In addition, described fluidic hardware 300 and described chuck 302 can make by certain size and shaping to be carried by automated system.

As shown in Figures 9 and 10, described chuck 302 can comprise shell or carrier frame 304 and be coupled to the lid component 306 of described shell 304.Described shell 304 has a shell or has along described Z axis towards reverse direction the height H extended betwixt ₂(as shown in figure 11) carrier side 303 and 305.As shown in Figure 9, described shell 304 comprises bridge members 324 at the loading end 316 of described fluidic hardware 300 and comprises substructure member 326 in the receiving end 318 that connects of described fluidic hardware 300.Described shell 304 is also included in a pair isolated leg prolongation 328 and 330 extended between described bridge members and substructure member 324 and 326.Described bridge members 324 extends and connects described leg prolongation 328 and 330 between described leg prolongation 328 and 330.Described bridge members 324 can comprise out the recess 321 (as shown in Figure 10) to the outside of described fluidic hardware 300.As shown in Figure 9, described leg prolongation 328 and 330 can have multiple clamping components 371-374 being configured to clamp the face, described pond 256 of described flow cell 200.

Still as shown in Figure 9, described fluidic hardware 300 can have the device window 315 passing completely through described chuck 302 along described Z axis.In the embodiment shown, described device window 315 is become with 330 frames by described bridge members 324, described lid component 306 and described leg prolongation 328 substantially.Described device window 315 comprises the recess 320 and 322 of spatial accommodation 308 and the described spatial accommodation 308 of multiple next-door neighbour.Described spatial accommodation 308 is configured to hold described flow cell 200.When described flow cell 200 is placed in described spatial accommodation 308, described flow cell 200 is exposed to the outside of described fluidic hardware 300, so as described flow cell 200 can in sight or along described shell surface 303 also have described shell surface 305 be directly engaged.Such as, face, described pond 258 (also as shown in figure 11) is along described Z axis towards the contrary direction relative to face, described pond 256.Face, described pond 256 can be seen by described imaging system or directly be engaged by another parts along described shell surface 303.Equally, face, described pond 258 can be seen by described imaging system or directly be engaged by another parts along described shell surface 305.

About Fig. 9 and 10, described lid component 306 can comprise the lid 340 and packing ring 342 that intercouple.Described packing ring 342 comprises the entrance and exit passage 346 and 344 (as shown in Figure 9) adjoined each other.In the embodiment shown, described lid 340 and described packing ring 342 are co-molded to integral structure.After formation, described lid 340 and described packing ring 342 can have different compressible character.Such as, in a particular embodiment, described packing ring 342 can comprise the material having more compressible character than the material of described lid 340.But, in substituting embodiment, described lid 340 and described packing ring 342 can be coupling in together with (as mechanically or use bonding agent) independent parts.In other embodiments, described lid 340 and described packing ring 342 can be different parts or the regions of single continuous print structure.

Described lid component 306 can be coupled to described shell 304 movably.Such as, described lid component 306 can be rotatably coupled to the described substructure member 326 of described shell 304.In such embodiment, described packing ring 342 is can around the turning axle R between installation site (as shown in Figure 9) and disengaged position (as shown in Figure 10) ₁rotate.Be coupled in the embodiment of described shell 304 movably at other wherein said lid components 306, described lid component 306 can be dismountable from described shell 304.Such as, when being connected to described shell 304, described dismountable lid component can in the installation site being similar to installation site as shown in Figure 9.When departing from described shell 304, described dismountable lid component can be completely removed at disengaged position.

Still as shown in Figure 10, described shell 304 can limit the chuck chamber 338 (Figure 10) that can enter when described lid component 306 is at described disengaged position.In some embodiments, identify that transmitter 336 can be placed in described chuck chamber 338.Described identification transmitter 336 is configured to the information of reception and registration about described flow cell 200 to reader.Such as, described identification transmitter 336 can be RFID label tag.When described flow cell 200 is inserted into described imaging system, the information that described identification transmitter 336 provides is passable, such as, the described sample in described flow cell 200, a large amount of described flow cells or sample, date of manufacture and/or pending mensuration scheme is identified.Described identification transmitter 336 also can pass on other information.

Figure 11 is the cross-sectional view of the described fluidic hardware 300 looked along described Y-axis.In some embodiments, described spatial accommodation 308 relative to described flow cell 200 press certain size make and shaping so that described flow cell 200 is maintained in described space, but at least some configuration can be floated wherein.As used herein, term " floats " and similar term comprises parts and is allowed at least in a direction (such as along described X, Y or Z axis) mobile limited distance.Such as, described flow cell 200 can have along the ability of described XY plane at described spatial accommodation 308 internal shift.Described flow cell 200 also can have the ability along the direction movement of described Z axis in described spatial accommodation 308.In addition, described flow cell 200 also can have the ability rotated a little in described spatial accommodation 308.In a particular embodiment, described shell 304 allows described flow cell 200 about any one of described X, Y and Z axis in described spatial accommodation 308 internal shift, movement with rotate a little.

In some embodiments, described spatial accommodation 308 also can be characterized as being such space: when described fluidic hardware 300 holds described flow cell 200, and described fluidic hardware 300 allows described flow cell 200 to move freely wherein.Therefore, the size of described spatial accommodation 308 can based on the position of reference field of described fluidic hardware 300 directly can engaging described flow cell 200.Described reference field can be the surface of described shell 304 or the described lid component 306 comprising described packing ring 342.Such as, Figure 11 illustrates multiple reference field 381-387.When described flow cell 200 is housed inside in described spatial accommodation 308, the reference field 383 of the reference field 381 and 382 that described clamping components 371 and 372 is respective and described packing ring 342 can limit the movement that described flow cell 200 exceeds predeterminated level.The described reference field 384 of described packing ring 342 and the reference field 385 of described bridge members 324 can limit the movement of described flow cell 200 along described XY plane.In addition, the reference field 386 and 387 of described bridge members 324 and described lid component 306 also can limit the movement of described flow cell 200 along described Z axis.But described reference field 381-387 is exemplary and described fluidic hardware 300 can have the reference field of the movement of other the described hydrogymnasium 200 of restriction.

Assemble described fluidic hardware 300, described flow cell 200 can be written into described spatial accommodation 308.Such as, described flow cell 200 can move forward towards described device window 315 along described shell surface 305.Described edge 234 (Fig. 5) can move forward between described clamping components 372 and 373 and described packing ring 342.Then, face, described pond 256 can rotate towards described clamping components 371-374, so that described clamping components 371-374 engages face, described pond 256.Then, described edge 232 (Fig. 5) towards described bridge members 324 and, more specifically, the described reference field 385 towards described bridge members 324 moves.In some embodiments, described bridge members 324 can deflect or bend to provide more for arranging the space of described pond end 246 (Fig. 5) thereon.When described flow cell 200 is written into described chuck 302, described shell 304 and described lid component 306 can clamp the periphery of described flow cell 200 effectively, so that described flow cell 200 is limited with only mobile in described spatial accommodation 308 inside.

In substituting embodiment, described pond end 246, first by described bridge members 324, then inserts location by described packing ring 342.In other implementations, described flow cell 200 can close on described shell surface 303.Described clamping components 371-374 can have the taper or the chamfered surface that allow described flow cell 200 to snap in the position in described spatial accommodation 308.

Before described flow cell 200 is loaded into, loading is rear or during being loaded into, described lid component 306 can be moved into described disengaged position, so that described identification transmitter 336 (Figure 10) can be placed in described chuck chamber 338 (Figure 10).When described packing ring 342 is in described installation site, described entrance and exit passage 346 and 344 can have precalculated position relative to described shell 304 and described spatial accommodation 308 and direction.Described packing ring 342 can be installed in the top of described flow cell 200 along the expose portion of described flow cell 200 (i.e. face, described pond 256).Described entrance and exit passage 346 and 344 usually can be aimed at described entrance and exit 224 and 222 (Fig. 5).

But, be to be noted that shown fluidic hardware 300 is a concrete embodiment, and in substituting embodiment, described fluidic hardware 300 can have different configurations.Such as, in substituting embodiment, described flow cell 200 can not along each outside being exposed to described fluidic hardware 300 of described shell surface 303 and 305.On the contrary, described flow cell 200 only can be exposed to described outside along one (such as described shell surface 303) of described shell surface.In addition, in substituting embodiment, described lid component 306 can not be rotatably coupled to described shell 304.Such as, described lid component 306 can be intactly dismountable.

Figure 12-15 illustrate according to substituting embodiment formed also can be used for Analytical system (as described in Analytical system 100 (Fig. 1) and as described in workstation 160 (Fig. 2)) fluidic hardware 900 and 920.Described fluidic hardware 900 and 920 can comprise the functional part similar with described fluidic hardware 300.Such as, as shown in the figure, in figs. 12 and 13, described fluidic hardware 900 can comprise chuck (or flow cell carrier) 902 and described flow cell 200.Described chuck 902 is configured to keep described flow cell 200 and is convenient to directed described flow cell 200 for imaging session.Described chuck 902 comprises shell 904 and is removably mounted to the lid component 906 of described shell 904.Described lid component 906 is in the installation site in Figure 12 and the disengaged position in Figure 13.

Still as shown in Figures 12 and 13, described fluidic hardware 900 can comprise the seal 910 of described inlet and outlet 222 and 224 (Figure 13) covering described flow cell 200.In some embodiments, described seal 910 is configured to be convenient to be retained in by fluid in described flow channel 206, so that the described sample in described flow channel 206 205 (Fig. 5) remains in fluid environment.But in some embodiments, described seal 910 can be configured to prevent unwanted material from entering described flow channel 206.As shown in Figures 12 and 13, described seal 910 is the monolithic adhesive tapes extended between described pond end 246 and 248 (Figure 13).Sponson 912 can extend away from described pond end 246.In substituting embodiment, described seal 910 can a slice adhesive tape (each each a slice adhesive tape of such as described inlet and outlet 222 and 224) or described seal 910 can be not just that other can cover the element of described inlet and outlet 222 and 224.Such as, described seal 910 can comprise stopper.

In some embodiments, when described fluidic hardware 900 is not installed to Analytical system, described seal 910 covers described inlet and outlet 222 and 224.Such as, when described fluidic hardware 900 is stored or transports or when in sample described flow cell 200 growth or generate time, described seal 910 can be used.In this case, described seal 910 can be fixed in described flow cell 200 and described shell 904, as shown in figure 13.More specifically, described seal 910 can be coupled to face, described pond 256 and extends along face, described pond 256 and cover described inlet and outlet 222 and 224.Described seal 910 also can be coupled to the substructure member 914 of described shell 904.Then, described lid component 906 can be moved to described installation site (as shown in figure 12), so that described seal 910 is sandwiched in described inlet and exports between 222 and 224 and described lid component 906.Described lid component 906 can be convenient to prevent described seal 910 from unexpectedly being removed.In substituting embodiment, described seal 910 can cover inlet and the exit passageway 916 and 918 of described lid component 906.

Figure 14 and Figure 15 illustrates described fluidic hardware 920, and described fluidic hardware 920 also can have the functional part similar with described fluidic hardware 300 and 900.As shown in the figure, described fluidic hardware 920 comprises chuck (or flow cell carrier) 922 and described flow cell 200.Described chuck 922 comprises shell 924 and is removably mounted to the lid component 925 of described shell 924.Show described lid component 925 and be only in installation site in Figure 14 and 15.Described shell 924 and described lid component 925 can with described shell 204 and 904 mentioned above and described lid component 206 and 906 similar.

But described shell 924 also can comprise Fin shape outshot 926 and 928.Described Fin shape outshot 926 and 928 press certain size make and shaping with by individual or robot device clamp, such as, when described fluidic hardware 920 is inserted into device support or removes from device support (not shown).In some embodiments, if described fluidic hardware 920 is not rationally placed, described Fin shape outshot 926 and 928 can stop described cap assemblies (not shown) to move to make-position.Described Fin shape outshot 926 and 928 can comprise the feeling function parts 927 and 929 being configured to be gripped by individual.In the embodiment shown, described Fin shape outshot 926 and 928 is placed in the receiving end 930 of described fluidic hardware 920.Described lid component 925 can extend between described Fin shape outshot.But described Fin shape outshot 926 and 928 can have other positions along described chuck 902.

Figure 16-24 shows the various functional parts of the fluidic hardware support 400 formed according to an embodiment.Figure 16 is the decomposed figure of described support 400.During assembling, described support 400 can be used to support described fluidic hardware 300 (Fig. 9) and described flow cell 200 (Fig. 5) by required orientation during imaging session.In addition, described support 400 can provide interface between described fluidic hardware 300 and described imaging system (not shown), and wherein said support 400 can be configured to guide fluid by described flow cell 200 and the heat energy providing or remove from described flow cell 200.Although described support 400 is shown as keeping described fluidic hardware 300, described support 400 can be configured to keep other jet apparatus, as lab on A Chip equipment or the flow cell not having chuck.

As shown in figure 16, described support 400 can comprise dismountable cap assemblies 404 and supporting construction 402.In some embodiments, described support 400 also can comprise plate structure 406 and moveable platform 408.Described plate structure 406 is operationally coupled to described cap assemblies 404 and comprises through opening 410 wherein.Equally, described platform 408 comprises through opening 412 wherein.The radiating module (or thermal cycler) 416 that described supporting construction 402 can comprise heating radiator 414 and be installed on described heating radiator 414.Described radiating module 416 comprises base part 418 and base 420.When assembling described support 400, described supporting construction 402, described platform 408 and described plate structure 406 are mutually stacking.On this point, described opening 412 makes by the size holding described base part 418 and shaping and described opening 410 makes by the size holding described base 420 and shaping.Upon assembly, described cap assemblies 404 can be operably coupled to described plate structure 406 and described supporting construction 402.

Figure 17 illustrates assembled support 400.In the embodiment shown, panel 424 is placed in the top of described plate structure 406 (Figure 16).As shown in FIG. 16 and 17, described cap assemblies 404 comprises the cover housing 435 being coupled to described plate structure 406.Described cover housing 435 can U-shaped in general, has a pair isolated shell leg 436 and 438 extended towards common direction.Described shell leg 436 and 438 can be rotatably coupled to described plate structure 406 at binding site 437 and 439 place.Described cover housing 435 also can be included in and to extend between described shell leg 436 and 438 and to connect the bridging portion 440 of described shell leg 436 and 438.So, described cap assemblies 404 can be configured to provide viewing space 442 (Figure 17).Described viewing space 442 can make by certain size and shaping to allow imaging len (not shown) with direction Dx (Figure 17) movement along described flow cell 200 and above described flow cell 200.

In the embodiment shown, described cap assemblies 404 is that relatively described plate structure 406 or supporting construction 402 are moveable between open position (as shown in figure 16) and make-position (as shown in figure 17).In described open position, the loading region 422 (as shown in figure 18) allowing to enter described support 400 is drawn back or retracted to described cap assemblies 404, so that described fluidic hardware 300 can be removed or be inserted into described loading region 422 by from described loading region 422.In described make-position, described cap assemblies 404 is installed to the top of described fluidic hardware 300.In a particular embodiment, described cap assemblies 404 is set up in described make-position and to be connected with the fluid of described fluidic hardware 300 and to be pressed in described supporting construction 402 by described flow cell 200.

As shown in figure 16, in some embodiments, described support 400 comprises coupling mechanism 450, so that keep described cap assemblies 404 in described make-position.Such as, described coupling mechanism 450 can comprise the element 452 (it comprises the button 453 being coupled to a pair lock opening 456 and 458) that operator controls.Described coupling mechanism 450 also comprises from a pair outstanding lock termination 454 and 455 of the matching surface 460 of described cover housing 435.Described cover housing 435 can be biased into described open position by spring element 464 and 466.When described cap assemblies 404 by individual or machine shift-in described in make-position time, described lock termination 454 and 455 be respectively inserted into described lock opening 456 and 458 and clamp described operator control element 452.Will by open position described in described cap assemblies 404 shift-in, described individual or machine by, such as, inwardly press described button 453 and start described button 453.Because described cover housing 435 is biased by described spring element 464 and 466, described cover housing 435 rotates away from described panel 424 (Figure 17) around described binding site 437 and 439.

In substituting embodiment, described coupling mechanism 450 can comprise other elements, so that described cap assemblies 404 is remained on described make-position.Such as, described lock termination 454 and 455 is replaceable is magnetic element or forms the element of interference engagement with opening.

Figure 18 is the skeleton view of the radiating module 416 of described supporting construction 402 and the isolated of described heating radiator 414.Described radiating module 416 can be configured to the temperature controlling described flow cell 200 in predetermined time section.Such as, described radiating module 416 can be configured to the temperature improving described flow cell 200, so that the DNA in described sample can sex change.In addition, described radiating module 416 can be configured to remove heat energy, thus reduces the temperature of described flow cell 200.As shown in the figure, described base 420 comprise by certain size make and shaping with the basal plane 430 engaged with described flow cell 200 (Fig. 5).Described basal plane 430 is towards the direction along described Z axis.Described base 420 also can comprise multiple alignment members 431-433 arranged around described basal plane 430.In the embodiment shown, described alignment members 431-433 has the fixed position relative to described basal plane 430.Described alignment members 431-433 has and is configured to accordingly engage described flow cell 200 and is convenient to arrange the reference field of described flow cell 200 for imaging.Such as, the described reference field of described alignment members 431-433 can towards the respective direction along described XY plane, and on this point, it can be configured to limit the movement of described flow cell 200 along described XY plane.Described supporting construction 402 can comprise described loading region 422 at least partially.Described loading region 422 partly can be limited by the described reference field of described basal plane 430 and described alignment members 431-433.

Figure 19 and 20 illustrates the alignment components 470 that can use together with described support 400 according to an embodiment.Figure 19 is the planimetric map of described support 400, and wherein said cover housing 435 is shown with cut-open view to illustrate described alignment components 470.Figure 20 is the skeleton view of described support 400, and wherein said cap assemblies 404 is in described open position.(in Figure 19 and 20 liang figure, described panel 424 (Figure 17) is deleted, only way for instructions.)

In Figure 19 and 20, described fluidic hardware 300 is written in described loading region 422.When described fluidic hardware 300 is written into, described flow cell 200 is placed to the upper and described alignment members 432,433 and 431 of described basal plane 430 (Figure 18) and is moved forwardly described recess 320,322 and 321 (Fig. 9 and 10) by described chuck 302.More specifically, along described shell surface 305 described device window 315 (Fig. 9) can by be greater than described basal plane 430 periphery size make and shaping.Thus, described chuck 302 or shell 304 can be allowed to fall described basal plane 430 around, but described flow cell 200 is prevented from falling described basal plane 430 side.So, the face, described pond 258 of described flow cell 200 can be crushed on described basal plane 430, so that described radiating module 416 can control the temperature of described flow cell 200.When described flow cell 200 is installed on described basal plane 430, the described reference field 381-383 (Figure 11) of described chuck 302 is crushed on face, described pond 256 (Figure 11).At this moment, the pond plane of the described flow cell 200 extended along described sample 205 can aim at the object plane of described imaging system substantially.

In the embodiment shown, when described fluidic hardware 300 is written into described loading region 422, the identification reader of described Analytical system can detect the information from described identification transmitter 336 (Figure 10).Such as, described support 400 can comprise the identification reader (not shown) of closing on described identification transmitter 336, being positioned at described plate structure 406.Identify that reading can occur in before described cap assemblies 404 is installed on described fluidic hardware 300.

With reference to Figure 19 and 20, described alignment components 470 comprises common cooperation with the described flow cell 200 of orientation and location with the various elements of imaging.Such as, described alignment components 470 comprises moveable registration arm 472 and is operably coupled to the actuator 474 of described registration arm 472.As shown in the figure, described actuator 474 comprises bar 476 and is coupled to the pin element 478 of described cover housing 435.In the embodiment shown, described bar 476 is around turning axle R ₂(Figure 19) rotatable.Described bar 476 can be L shape, has the first prolongation 480 being configured to engage described pin element 478 and the second prolongation 482 being configured to engage described registration arm 472.Described registration arm 472 is also around turning axle R ₃(Figure 19) rotatable and comprise the finger piece 484 with end of engagement 486.Described alignment components 470 also comprises the biasing element 490 (such as volute spring) engaging described finger piece 484.Described end of engagement 486 is configured to the described chuck 302 engaging described fluidic hardware 300.In substituting embodiment, described end of engagement 486 can be configured to directly engage described flow cell 200.

Described alignment components 470 is in engagement and arranges and be in fig. 20 and draw back layout in Figure 19.When drawing back layout described in described alignment components 470 is in, described registration arm 472 is in retracted position and when described alignment components 470 is in described engagement layout, described registration arm 472 is in offset position.Will aim at described flow cell 200 in described loading region 422, described alignment components 470 changes to described engagement layout from described layout of drawing back.Such as, when described cover housing 435 moves to described open position (as shown in figure 20), described pin element 478 engages described first prolongation 480 of described bar 476, makes described bar 476 by counterclockwise around described axle R ₂rotate (as shown in figure 19).Described cover housing 435 is maintained at described open position by described spring element 464 and 466 (Figure 16).When described bar 476 rotates, described second prolongation 482 is around described axle R ₂rotate and engage described registration arm 472.Described registration arm 472 is in the direction of the clock around described axle R ₃rotate (as shown in figure 19).When described registration arm 472 rotates, described registration arm 472 is moved to described retracted position.When moving on to described retracted position, described end of engagement 486 moves away from the described reference field of described alignment members 431-433.

Make described alignment components 470 be altered to described engagement layout from described layout of drawing back, described cover housing 435 can rotate towards described fluidic hardware 300 and be installed to the top of described flow cell 200.When described cover housing 435 moves towards described fluidic hardware 300, described pin element 478 rotates away from described first prolongation 480 of described bar 476.When described second prolongation 482 moves away from described registration arm 472, the potential energy being stored in described biasing element 490 can cause described registration arm 472 by counterclockwise rotating, and described end of engagement 486 is pressed on described chuck 302.On this point, described registration arm 472 is moved to described offset position.When being moved to described offset position, described end of engagement 486 moves towards the described reference field of described alignment members 431-433.

Figure 21 is the enlarged drawing of the fluidic hardware 300 described in described loading region 422 when the described end of engagement 486 of described registration arm 472 is pressed on described chuck 302.Described end of engagement 486 can be configured to move in described XY plane between described retracted position and described offset position.When described end of engagement 486 to move towards described offset position and is pressed on described chuck 302, described end of engagement 486 provides the power F for described chuck 302 _xY.Described chuck 302 can along described XY plane displacement and/or the described reference field described flow cell 200 being pressed in described alignment members 431-433.Described power F _xYthere is X component and Y-component.Described flow cell 200 can be pressed in described alignment members 431 by described X component, and described flow cell 200 can be pressed in described alignment members 432 and 433 by described Y-component.On this point, described alignment members 431 can stop described flow cell 200 along the movement of described X-direction, and described alignment members 432 and 433 can stop described flow cell 200 along the movement of described Y direction.

Be altered to before described engagement arranges in described alignment components 470, the described inlet of described lid component 306 and exit passageway 346 and 344 can respectively with the described inlet of described flow cell 200 with export 224 and 222 (Fig. 7) rough alignment.After described alignment components 470 is altered to described engagement layout, described inlet and exit passageway 346 and 344 and described inlet and export 224 and 222 effectively (or operationally) and aim at, so that fluid can flow through wherein effectively.

Therefore, described cap assemblies 404 can be operably coupled to described alignment components 470, so that a step or action can make described alignment components 470 engage described fluidic hardware 300.More specifically, when above described cap assemblies 404 is installed to the described device being in described make-position, described actuator 474 moves described registration arm 472 to described offset position.At described offset position, described registration arm 472 supports the described reference field of described flow cell 200 against the described alignment members 431-433 be in a fixed position along described XY plane.When described cap assemblies 404 is in described make-position, described viewing space 442 (Figure 17) can be placed in the top of described flow cell 200, so that imaging len can move along described flow cell 200 with flow channel described in imaging 206.Along with described cap assemblies 404 moves to described open position, described registration arm 472 is moved to described retracted position by described actuator 474.But in the embodiment shown, when described registration arm 472 is retracted, described flow cell 200 remains on original position.Therefore, described flow cell 200 can be relocatable relative to various element.Such as, when described lid component 306 is in described installation site, described flow cell 200 can be relative to described lid component 306 and described packing ring 342 relocatable.Described flow cell 200 also can be relative to described cap assemblies 404 and described basal plane 430 relocatable.

In some embodiments, described alignment components 470 and described cap assemblies 404 can by predetermined sequential operation.Such as, in a particular embodiment, before described cap assemblies 404 arrives described make-position, described registration arm 472 is configured to support described flow cell 200 against the described alignment members 431-433 being in described fixed position.When described cap assemblies 404 arrives described make-position, described cap assemblies 404 can be convenient to described flow cell 200 to be pressed on described basal plane 430 and also by described inlet and exit passageway 346 and 344 be pressed in described inlet and outlet 224 and 222.Usually, at described basal plane 430 after flow cell 200 described in z dimension inner position, described alignment components 470 can be configured to flow cell 200 described in x and y dimension inner position.In addition, alignment components can be configured to first then flow cell 200 described in z dimension inner position in x and y dimension.Therefore, the aligning in x, y and z dimension can respond single step that user takes or action one after the other and occur in sequence by various.

In substituting embodiment, described alignment components 470 can not be operably coupled to described cap assemblies 404 as described above.On the contrary, described alignment components 470 and described cap assemblies 404 can operate independently of one another.On this point, perhaps individual needs to perform multiple step to aim at described flow cell 200 and the jet ground described flow cell 200 that is coupled.Such as, described alignment components 470 can be started separately by individual, thus mobile described registration arm 472 is to aim at described flow cell 200.After described flow cell 200 is aligned, described individual can reduce described cap assemblies 404 again on described flow cell 200.In addition, described alignment components 470 can comprise the extra and/or miscellaneous part except those parts above-mentioned.

Figure 22 is the isolated skeleton view of the described cap assemblies 404 being in described make-position.Figure 22 illustrates the size in described viewing space 442.As shown in the figure, described cover housing 435 can have top surface 492.Described viewing space 442 can have the degree of depth D recorded to described fluidic hardware 300 or described flow cell 200 from described top surface 492 _p.Described viewing space 442 also can have the width W recorded along described Y-axis ₆with the length L recorded along described X-axis ₆.The length, width and height in described viewing space 442 can make by certain size, so that imaging len (not shown) can through wherein moving above described flow cell 200.More specifically, imaging len can enter described viewing space 442 by inspection hole 443 and direction along described X-axis is moved above described flow cell 200.

Figure 23 is the cross-sectional view of the described cap assemblies 404 got along the line 23-23 in Figure 22.In the embodiment shown, described cap assemblies 404 can comprise multiple compression arm 494 and 496.Described compression arm 494 and 496 is configured to the force of compression F of the described shell surface 303 provided respectively for described fluidic hardware 300 _c1and F _c2.In the embodiment shown, described compression arm 494 and 496 is pressed onto on described chuck 302.But in substituting embodiment, described compression arm 494 and 496 can be pressed onto on described flow cell 200.

Described force of compression F _c1and F _c2press the described shell 304 of described fluidic hardware 300, thus the face, described pond 256 (Fig. 9) of described flow cell 200 is pressed onto on described radiating module 416.On this point, described flow cell 200 can keep close contact with described basal plane 430, in order to transferring heat energy betwixt.In the embodiment shown, described compression arm 494 and 496 operates independently of one another.Such as, each of described compression arm 494 and 496 is operably coupled to respective Compress Spring 495 and 497.

As shown in figure 23, described compression arm 494 and 496 extends towards described viewing space 442 and described loading region 422.When described cap assemblies 404 moves on to described make-position, the engageable described shell surface 303 of described compression arm 494 and 496.Because described compression arm 494 and 496 is pressed onto on described shell surface 303, the resistance from described shell surface 303 can make described compression arm 494 and 496 around axle R ₄and R ₅rotate.Each rotation of resisting respective compression arm in described Compress Spring 495 and 497, thus the corresponding force of compression F for described shell surface 303 is provided _c.Therefore, described compression arm 494 and 496 relative to each other independent bias.

Figure 24 is the isolated skeleton view of the stream sending component 500 of described cap assemblies 404 (Figure 16).Described stream sending component 500 comprises manifold body 502 and upstream and downstream flow line 504 and 506.As shown in figure 16, described manifold body 502 can extend between described shell leg 436 and 438.Turn back to Figure 24, described flow line 504 and 506 respectively body port 508 and 510 by mechanically and jet be coupled to described manifold body 502.Described flow line 504 and 506 also comprises the pipe end 514 and 516 of described inlet and the exit passageway 346 and 344 being configured to be inserted into described packing ring 342.

As shown in figure 24, described stream sending component 500 relative to described packing ring 342 in installation site.In described installation site, described pipe end 514 and 516 is inserted into described entrance and exit passage 346 and 344 respectively, so that fluid can flow through described flow cell 200.In addition, in described installation site, described packing ring 342 (Fig. 9) can be pressed on described flow cell 200 by described stream sending component 500, is sealed effectively so that fluid connects.For this reason, described stream sending component 500 can comprise bias spring 520 and 522.Described bias spring 520 and 522 is configured to be pressed onto the inside of described cover housing 435 (Figure 16) and provides the power F for described packing ring 342 _c3.Described coupling mechanism 450 (Figure 16) can so that keep the sealing to described packing ring 342.

Therefore, described cap assemblies 404 can be pressed onto on the described shell 304 of described fluidic hardware 300 three independent compression point.More specifically, when being engaged by described pipe end 514 and 516, described packing ring 342 can form the first compression point P ₁(as shown in figure 24), and described compression arm 494 and 496 can second and the 3rd compression point P ₂and P ₃(as shown in figure 23) described fluidic hardware 300 is contacted.As shown in figs. 22-24, described three compression point P ₁-P ₃distribute around described flow cell 200.In addition, described cap assemblies 404 is at described compression point P ₁-P ₃described force of compression F is provided independently _c1-F _c3.On this point, described cap assemblies 404 can be configured to provide the substantial uniform force of compression for described fluidic hardware 300, so that described flow cell 200 is pressed onto on described basal plane 430 equably and jet connection is sealed with anti-leak.

Figure 25 is a kind of block diagram of localization method 530 of the fluidic hardware for sample analysis.Described method 530 is included on basal plane and arranges dismountable fluidic hardware (see 532).Described fluidic hardware can be similar to fluidic hardware 300 mentioned above.Such as, described fluidic hardware can comprise spatial accommodation, be positioned at flow cell and the packing ring of described spatial accommodation.Described flow cell can extend along the object plane in described spatial accommodation and can be relocatable relative to the described packing ring in described object plane.Described method 530 to be also included in described spatial accommodation but on described basal plane mobile described flow cell so that the entrance and exit passage rough alignment (see 534) of the entrance and exit of described flow cell and described packing ring.Mobile operation 534 can comprise startup localizer arm to be pressed onto in described alignment members by described flow cell.

Figure 26 is a kind of block diagram of localization method 540 of the fluidic hardware for sample analysis.Described fluidic hardware can be similar to fluidic hardware 300 mentioned above.Described method 540 comprises providing and has device outer case, comprises spatial accommodation and be positioned at the fluidic hardware (see 542) of relocatable flow cell of described spatial accommodation.Described device outer case can comprise the recess being close to described spatial accommodation.Described method also comprises described fluidic hardware to be arranged on and has (see 544) in the supporting construction of alignment members.Described alignment members is inserted into by corresponding recess.In addition, described method 540 can be included in mobile described flow cell (see 546) in described spatial accommodation.When described flow cell moves in described spatial accommodation, described alignment members can engage the edge of described flow cell.Mobile operation 546 can comprise startup localizer arm to be pressed onto in described alignment members by described flow cell.

Figure 27 illustrates a kind of block diagram for the method 550 relative to orthogonal X, Y and Z axis localizing sample region.Described method 550 comprises provides alignment components (see 552).Described alignment components can be similar to alignment components 470 mentioned above.More specifically, described alignment components can comprise the moveable registration arm with end of engagement.Described registration arm can be moveable between retraction and offset position.Described method 550 also comprises and is arranged on the basal plane in the direction along described Z axis by fluidic hardware and is arranged between multiple reference field towards the respective direction along XY plane (see 554).In addition, described method 550 also comprises mobile described registration arm to described offset position (see 556).Described device can be pressed onto on described reference field by described registration arm, so that described device is maintained at fixing position.

Figure 28-37 illustrates the various functional parts of fluid storage system 1000 (Figure 28).Described storage system 1000 is configured to store the various fluid that can use between predetermined test period and the temperature regulating described fluid.Described storage system 1000 uses by described workstation 160 (Fig. 2) and is closed by described shell 162 (Fig. 3).As shown in figure 28, described storage system 1000 comprises shell 1002, and described shell 1002 has the base shell (or first shell) 1004 and top casing (or second housing) 1006 that are coupled and limit system chamber 1008 betwixt.Described shell 1002 also can comprise system door 1010, and described system door 1010 is configured to open and provides the approach entering described system chamber 1008.Still as shown in the figure, described storage system 1000 can comprise the temperature-controlling module 1012 at the rear portion being coupled to described shell 1002 and the elevator drives motor 1014 be arranged in described top casing 1006.

Figure 29 is the cross-sectional side view of described storage system 1000 and illustrates in greater detail described system chamber 1008.Described storage system 1000 also can comprise reacted constituent pallet (or reacted constituent storage unit) 1020 and pipette assembly 1022 with the fluid comprising elevating mechanism 1024.Described pallet 1020 is configured to support multiple pipeline for depositing fluid or container.Described elevating mechanism 1024 comprises described drive motor 1014 and is configured to pipette along fluid described in described Z axis way moving the parts of assembly 1022.In Figure 29, described pallet 1020 is positioned at fluid and pipettes position, such as, so that the fluid that described pallet 1020 keeps can be pipetted and be transported to, fluidic hardware, in order to perform required reaction or the described flow channel in order to rinse described fluidic hardware.

Still as shown in the figure, described temperature-controlling module 1012 can protrude into described system chamber 1008.Described temperature-controlling module 1012 is configured to control or regulate the temperature in described system chamber 1008.In the embodiment shown, described temperature-controlling module 1012 comprises thermoelectric cooling (TEC) assembly.

Figure 30 pipettes the skeleton view of assembly 1022 described in being.As shown in the figure, pipette assembly 1022 described in and can comprise a pair relative guide rail 1032 and 1034.Described relative guide rail 1032 and 1034 is configured to receive and guides described pallet 1020 to pipette position to described fluid, as shown in figure 29.Described guide rail 1032 and 1034 can comprise along described guide rail 1032 and 1034 outstanding functional part extending longitudinally or ridge 1035.Described guide rail 1032 and 1034 is configured to be fixed in described base shell 1004 (Figure 28).The described assembly 1022 that pipettes also comprises the brace summer (or column) 1036 and 1038 extended along described Z-direction.The described guide plate 1040 pipetting assembly can with the distance D raised _zbe coupled to described brace summer 1036 and 1038 and stretch out along described XY plane thus.In the embodiment shown, described guide plate 1040 is fixed in described brace summer 1036 and 1038.

Described elevating mechanism 1024 comprises structural supports 1041 and 1042, the leading screw 1044 extended between described structural supports 1041 and 1042 and comprise the objective table assembly 1046 of delivery platform 1048.Described structural supports 1041 and 1042 is fixed in the two ends of described brace summer 1036 and 1038 and is configured in operational process, support described elevating mechanism 1024.The screw thread of described leading screw 1044 is operably coupled to described objective table assembly 1046, and when rotating with the described leading screw 1044 of box lunch, described objective table assembly 1046 moves (being represented by double-head arrow) along described Z axis by rectilinear direction.

Described delivery platform 1048 is configured to supporting suction pipe 1050 array.Described suction pipe 1050 can be communicated with the systems pumps (not shown) fluid being configured to guide fluid to flow through described suction pipe 1050.As shown in the figure, described suction pipe 1050 comprises the distal portions 1052 in the composition hole 1060 (as shown in figure 31) being configured to be inserted into described pallet 1020.Described distal portions 1052 extends through the respective openings 1053 of described guide plate 1040.

Described elevating mechanism 1024 is configured to drawing back level and transferring mobile described suction pipe 1050 between level.Described transfer level time (shown in Figure 50 and 51), the described distal portions 1052 of described suction pipe 1050 is inserted into described composition hole 1060 with from wherein pipetting fluid.Described draw back level time, described distal portions 1052 is completely removed described pallet 1020, so that described pallet 1020 can be moved out of described system chamber 1008 (Figure 28), and does not damage described suction pipe 1050 or described pallet 1020.More specifically, when described drive motor 1014 rotates described leading screw 1044, described objective table assembly 1046 moves along described Z axis with the direction determined by the sense of rotation of described leading screw 1044.Therefore, described transmission platform 1048 moves along described Z axis, supports described suction pipe 1050 simultaneously.If described delivery platform 1048 advances towards described guide plate 1040, described distal portions 1052 slides towards described pallet 1020 through the respective openings 1053 of described guide plate 1040.Described guide plate 1040 is configured to prevent distal portions 1052 from becoming before it is inserted into described composition hole 1060 and no longer aims at described composition hole 1060.When described objective table assembly 1046 is moved apart described guide plate 1040 by described elevating mechanism 1024, distance (Δ Z) between described delivery platform 1048 and described guide plate 1040 becomes large, until described distal portions 1052 is drawn back from the described composition hole 1060 of described pallet 1020.

Figure 30 illustrates the attachment feature for running described elevating mechanism 1024.Such as, described objective table assembly 1046 also can comprise guide pin 1058 (still as shown in figure 29), and described guide pin 1058 is fixed on described delivery platform 1048 and extends from described delivery platform 1048 with the direction being parallel to described suction pipe 1050.Described guide pin 1058 also extends through the corresponding opening 1053 of described guide plate 1040.In the embodiment shown, the distance that described guide pin 1058 extends than described suction pipe 1050 is larger, so that before described suction pipe 1050 is inserted into described composition hole 1060, described guide pin 1058 arrives described pallet 1020.Therefore, if described pallet 1020 is not aimed at relative to described suction pipe 1050, described guide pin 1058 can engage described pallet 1020 and adjust the position of described pallet 1020, so that described composition hole 1060 is correct before described suction pipe 1050 is inserted into wherein aim at corresponding suction pipe 1050.

Except mentioned above, described in pipette assembly 1022 and can comprise position transducer 1062 and alignment sensor (not shown).Described position transducer 1062 is configured to the label 1063 (as shown in figure 34) receiving described pallet 1020, with determine described pallet 1020 to be present in described system chamber 1008 (Figure 28) and at least rough alignment in order to receive described suction pipe 1050.Described alignment sensor can detect the label 1064 of described objective table assembly 1046, to determine the level of described objective table assembly 1046.If described label 1064 does not also reach threshold level along described Z axis, described alignment sensor can be linked up with described workstation 160 (or other Analytical system), to notify that pallet 1020 described in user does not prepare to remove.Described workstation 160 also can prevent user from opening described system door 1010.

In addition, when the described distal portions 1052 of described suction pipe 1050 is inserted into described composition hole 1060 at first, described suction pipe 1050 can pierce through the protective foil covering described composition hole 1060.In some cases, described paper tinsel can clamp described suction pipe 1050.When described suction pipe 1050 is drawn back from corresponding composition hole 1060 subsequently, the clamping of described protective foil can lift to power described pallet 1020 together.But in the embodiment shown, described ridge 1035 is configured to holding tray base 1070 (Figure 31) and prevents described basetray 1070 to be lifted along the direction of described Z axis.Such as, described ridge 1035 can clamp the lip 1071 of described basetray 1070.

Figure 31-34 illustrates the different views of described pallet 1020.Described pallet 1020 is configured to support multiple composition hole 1060.Described composition hole 1060 can comprise various reacted constituent; such as, but be not limited only to one or more samples, polymerase, primer, denaturant, the linearization potpourri for linearization DNA, the enzyme being applicable to concrete mensuration (as bunch amplification or a SBS), nucleotide, cleavage mixture, oxidation protection agent and other reagent.In some embodiments, described pallet 1020 can hold the required all fluids of the predetermined mensuration of execution.In a particular embodiment, described pallet 1020 can be contained in flow cell and generate sample (such as DNA bunch) and perform the required all reacted constituents of sample analysis (such as SBS).Described mensuration can be performed when not removing or change any described composition hole 1060.

Described composition hole 1060 comprises quadratic component hole 1060A (as shown in Figure 35-36) and tubulose composition hole 1060B (as shown in figure 37).Described pallet 1020 comprises basetray 1070 and is coupled to the pallet cover 1072 of described basetray 1070.As shown in figs. 31 and 32, described pallet cover 1072 comprises handle 1074, described handle 1074 press certain size make and shaping, with the user's handling by described pallet 1020.Described pallet cover 1072 also can comprise and grips recess 1076, and described gripping recess 1076 makes by the size of the one or more fingers holding described user and shaping.

As shown in figs. 31 and 32, described pallet cover 1072 can comprise the pore 1080 that multiple with corresponding composition hole 1060 is aimed at.Described pore 1080 is plastic to guide described suction pipe 1050 (exemplary suction pipe 1050 as shown in figure 31) to enter corresponding composition hole 1060.As shown in figure 32, described pallet cover 1072 also comprises pin hole 1082, and described pin hole 1082 makes by the size holding described guide pin 1058 and shaping.Described guide pin 1058 be configured to when described guide pin 1058 with non-aligned mode near and enter described pin hole 1082 time, the slight adjustment of the position to described pallet 1020 is provided.Still as shown in the figure, described pallet 1020 can comprise the identification label 1084 on the surface along described pallet cover 1072.Described identification label 1084 is configured to be read device and detects, to provide the information of the fluid held about described composition hole 1060 to described user.

As seen in figs. 33 and 34, described pore 1080 is limited by the edge 1086 that the surface 1073 from described pallet cover 1072 is outstanding at least in part.Described edge 1086, from described surperficial 1073 outstanding little segment distances, mixes accidentally to prevent the fluid accidentally deposited on described pallet cover 1072.Equally, described identification label 1084 can be attached to the bossing 1088 of described pallet cover 1072.Described bossing 1088 also can protect described identification label 1084 accidentally to touch fluid.

Figure 35 shows the cross-sectional side view of described composition hole 1060A, and Figure 36 shows the bottom perspective view of described composition hole 1060A.As shown in the figure, the described composition hole 1060A storage 1090 (Figure 35) that comprises relative first end and the second end 1091 and 1092 and extend betwixt.Described storage 1090 has degree of depth D _r(Figure 35), described degree of depth D _rextend to described first end 1091 along with described storage 1090 from described second end 1092 and increase.Described composition hole 1060A is configured to the darker part described suction pipe 1050 being contained in described storage 1090.As shown in figure 36, described composition hole 1060A comprises multiple outshot 1094 being configured to lean against on the surface of described basetray 1070 along outside surface.

Figure 37 is the skeleton view of described composition hole 1060B.As shown in the figure, described composition hole 1060B also can comprise the outshot 1096 of multiple outside surface around described composition hole 1060B.Described composition hole 1060B longitudinally axle 1097 extends and has along with described composition hole 1060B extends lengthwise into bottom 1098 and tapered profile.Described bottom 1098 can have surface flat substantially.

Figure 61 illustrates that one is used for carrying out biological or chemical and analyzes method for measuring 960.In some embodiments, described mensuration can comprise sample generation scheme and sample analysis scheme.Such as, described sample generates scheme and can comprise and generate DNA bunch and described sample analysis scheme by bridge amplification and can comprise the synthesis limit, limit order-checking (SBS) utilizing described DNA bunch and analyze.Sample generate and sample analysis operation can common Analytical system (as described in Analytical system 100 or as described in workstation 160) in carry out and as described between operation without the need to user intervention.Such as, fluidic hardware can be loaded into described Analytical system by user.Described Analytical system automatically can generate the sample for analyzing and carry out performing the step of described analysis.

About Figure 61, described method 960 is included between the fluidic hardware with sample area and the reacted constituent storage unit containing multiple differential responses composition and sets up fluid connection (see 962).Described reacted constituent can be configured to measure for carrying out one or more.Described fluidic hardware can be, such as, and fluidic hardware 300 mentioned above or flow cell 200.In some embodiments, described sample area comprises multiple reacted constituent (such as primer) be fixed thereon.Described storage unit can be, such as, and storage unit 1020 mentioned above.Described reacted constituent can comprise the sample generating component be configured to for generating described sample and the sample analysis composition be configured to for analyzing described sample.In a particular embodiment, described sample generating component comprises the reacted constituent for performing bridge amplification mentioned above.In addition, in a particular embodiment, described sample analysis composition comprises the reacted constituent analyzed for performing SBS mentioned above.

The described sample area that described method 960 is also included in described fluidic hardware generates sample (see 964).Generating run 962 can comprise make different sample generating components flow into described sample area and the reaction conditions controlling described sample area to generate described sample.Such as, thermal cycler can be used so that nucleic acid hybridization.But, if necessary, can isothermal method be used.In addition, the flow velocity of described fluid can be controlled, to allow hybridization or the chemical reaction needed for other.In a particular embodiment, described generating run 962 comprises and carries out multiple bridge amplification circulation, to generate DNA bunch.

The exemplary arrangement of bridge amplification can comprise following step.Flow cell is set up and is communicated with reacted constituent storage unit fluid.Described flow cell comprises the one or more surfaces that attached to primer pair.Solution contact solid carrier containing not homotactic target nucleic acid potpourri.Described target nucleic acid can have common priming site, and the described primer pair on described priming site and described flow cell surface is complementary, so that described target nucleic acid is in conjunction with the first primer of the described primer pair on described flow cell surface.Extension solution containing polymerase and nucleotide can be introduced into described flow cell, to be formed by the extension of described first primer with the first amplified production of complementary target.Described extension solution can be removed and replace with denaturing soln.Described denaturing soln can comprise chemical denaturant, as NaOH and/or formamide.Consequent Denaturing discharges the former chain of described target nucleic acid, and it then can by removing described denaturing soln and removing from described flow cell with the replacement of described extension solution.Under described extension solution exists, described first amplified production being adhered to described carrier then can with the second primer hybridization of described primer pair adhering to described flow cell surface, and comprise can extending to form by described second primer with the second amplified production of the nucleotide sequence of the adhesion of described first amplified production complementation.The discrete location that described denaturing soln and the conveying repeatedly extending solution can be used on described flow cell surface forms target nucleic acid bunch.Although such scheme utilizes chemical modification exemplarily, understanding also can be carried out thermal denaturation to similar primer and target nucleic acid.The further describing of amplification method that can be used for producing immobilized nucleic acids molecular cluster is provided in, such as U.S. Patent number 7,115,400; US publication 2005/0100900, WO00/18957 or WO98/44151 (wherein each be incorporated herein by reference).

Described method 960 also comprises the described sample (see 966) analyzing described sample area.Usually, described analysis operation 966 can comprise any detectable feature detecting described sample area.In a particular embodiment, described analysis operation 966 comprises and makes at least one sample analysis composition flow into described sample area.Described sample analysis composition can react with described sample, to provide the optically detectable signal of instruction dependent event (or required reaction).Such as, described sample analysis composition can be the fluorescently-labeled nucleotide used in SBS analytic process.When exciting light incides on the described sample mixing fluorescently-labeled nucleotide wherein, described nucleotide can send the light signal of instruction nucleotide type (A, G, C or T), and described imaging system can detect described light signal.

A kind of useful especially SBS scheme utilizes the modified ribonucleotide having removable 3 ' end and close, such as, as described in WO04/018497, U.S. 2007/0166705A1 and U.S. 7,057,026 (wherein each be incorporated herein by reference).SBS reagent can be transported to by iterative cycles the flow cell that attached to target nucleic acid, such as, due to the bridge amplification scheme set forth above.Described nucleic acid bunch can use linearization solution to be converted to the form of strand.Described linearization solution can comprise, such as, and the restriction enzyme of a chain of energy cracking each bunch.Concerning restriction enzyme or nickase, other cleavage method can be selected as one, comprise especially chemical cracking (such as with periodate cleavage diol bond), by with restriction endonuclease (such as NEB, Ipswich, MA, the item number that USA provides is ' USER ' of M5505S) cracking, by being exposed to the abasic site cracking that heat or alkali carry out, mix the cracking of the ribonucleotide of the amplified production be made up of deoxyribonucleotide in addition, the photodestruction of peptide linker or cracking.After linearization step, in sequencing primer hybridization under the condition of target nucleic acid to be checked order, described sequencing primer can be delivered to described flow cell.

Then described flow cell can contact under certain condition with containing holding closed modified ribonucleotide and fluorescently-labeled SBS to extend reagent with removable 3 ', is extended by the primer of hybridization to each target nucleic acid to be added by mononucleotide.A mononucleotide is only had to be added into each primer, because once described modified ribonucleotide has been incorporated into the ever-increasing polynucleotide chain with the template area complementation of checking order, just do not have the free 3 '-OH group that further sequence can be guided to extend, therefore polymerase can not add more nucleotide.Described SBS extends reagent and can remove and replace the scanning reagent to contain the composition protecting described sample under radiation excitation.Description for the Exemplary compositions scanning reagent sees the U.S. and announces US2008/0280773A1 and United States serial 13/018,255 (wherein each be incorporated herein by reference).Then, under the existence of scanning reagent, the nucleic acid that fluoroscopic examination extends.Once fluorescence be detected, described 3 ' end is closed and the deblocking reagent being applicable to blocking groups used can be used to remove.The description that can be used for the exemplary deblocking reagent of respective blocking groups sees WO04018497, US2007/0166705A1 and US7057026 (wherein each be incorporated herein by reference).Described deblocking reagent can be washed off, and allow target nucleic acid hybridize to the extension primer containing 3 ' OH group, described 3 ' OH group can add more nucleotide now.Therefore, can repeat to add extend reagent, the circulation that closed reagent conciliate by scanning reagent and alternatively described step one or more between cleaning, until the sequence needed for obtaining.When described modified ribonucleotide each has different mark (alkali that its known correspondence is concrete) adhering to it, eachly can recycle single reagent supplying step that extends and carry out above-mentioned circulation.Described different mark be convenient to distinguish each mix step during the alkali that adds.Alternatively, each circulation can comprise independent extension reagent supplying step, succeeded by independent scanning reagent supplying step and detection, in this case, the two or more of described nucleotide can have identical mark, and can be distinguished based on known sequence of batching products.

Continue the example of flow cell amplifying nucleic acid bunch, described nucleic acid can be further processed with by being called that the method for end pairing order-checking obtains the second reading from the other end.The description of end pairing sequence measurement sees PCT and announces in WO07010252, PCT application sequence number PCTGB2007/003798 and U.S. Patent Application Publication US2009/0088327 (wherein each be incorporated to herein with quoting method).In one embodiment, a series of step can be performed as follows: as above set forth generate bunch, as above set forth linearization, still as above set forth hybridization first sequencing primer and carry out the extension of iterative cycles, scanning and deblocking, still as above set forth by synthesize on flow cell surface as described in complementary copy " inversion " as described in target nucleic acid, linearization synthesize chain again, hybridize the first sequencing primer and carry out the extension of iterative cycles, scanning and deblocking.Described inversion step can as above set forth and carried out for single-cycle bridge amplification by delivery of therapeutic agents.

Although described analysis operation about concrete SBS scheme as above exemplarily, can carry out understanding other schemes being used for checking order other kinds analysis of molecules any one as required.In view of enlightenment in this paper and the general knowledge about specific analytical method, for the suitable modifications adapting to the described apparatus and method that different analyses is done will be apparent.

In some embodiments, described method 960 is configured to carry out under minimum user intervention.Described generating run and analysis operation 964 and 966 can be carried out in an automatic fashion by Analytical system.Such as, in some cases, user only can be loaded into described fluidic hardware and described storage unit, and starts described Analytical system to execute a method described 960.In some embodiments, during described generating run and analysis operation 964 and 966, described storage unit and described fluidic hardware keep fluid to be communicated with from described generating run and during whole described analysis operation, until described sample is fully analyzed.In other words, described fluidic hardware and described storage unit can from before described sample generates until all keep fluid to be communicated with after described sample analysis.In some embodiments, described fluidic hardware is constantly supported by described device support from described generating run and during whole described analysis operation, until described sample is fully analyzed.During during this period of time, described device support and imaging len can relative to each other movements automatically.When described device support and described imaging len relative to each other move automatically, described storage unit and described fluidic hardware can keep fluid to be communicated with.In some embodiments, described Analytical system is included in workstation shell and described generating run and analysis operation 964 and 966 carry out independently in workstation shell.

Figure 38 is the schematic diagram of the optical imaging system 600 according to an embodiment formation.Described imaging system 600 comprises optical module 602, light source (or exciting light) module or assembly 604, the flow cell 606 with sample area 608 and imaging detector 610 and 612.Described light source module 604 comprises the first and second excitation sources 614 and 616 being configured to irradiate described sample area 608 with different excitation spectrums.In a particular embodiment, described first and second excitation sources 614 and 616 comprise the first and second semiconductor light sources (SLS).SLS can comprise light emitting diode (LED) or laser diode.But, in other implementations, can other light sources be used, as laser or arc lamp.Described first and second SLS can have the fixed position relative to described optical module 602.

As shown in the figure, described optical module 602 can comprise multiple optics.Such as, described optical module 602 can comprise lens 623-625, utilizing emitted light filtrator 631, exciting light filtrator 635 and mirror 644 and 642.Described optical module 602 can have extra for guiding described exciting light and/or described radiative optics (as lens, utilizing emitted light or exciting light filtrator, mirror etc.), and its Reference numeral is summarised as 621.Described multiple optics is configured to for following at least one: (a) guides described exciting light to collect the utilizing emitted light from described sample area 608 towards the described sample area 608 of described flow cell 606 or (b).Still as shown in the figure, described imaging system 600 also can comprise the flow system 652 be communicated with described flow cell 606 fluid and the system controller 654 being communicatively coupled to described first and second excitation sources 614 and 616 and described flow system 652.Described controller 654 is configured to start described flow system 652 to make reagent flow into described sample area 608 and after a predetermined time period, to activate described first and second SLS.

Such as, Figure 60 illustrates that one is used for carrying out biological or chemical and analyzes method for measuring 900.In a particular embodiment, described mensuration can comprise synthesis limit, limit order-checking (SBS) scheme.Described method 900 comprises the flow channel (see 902) making reagent flow through flow cell.Described flow cell can have sample area, and described sample area comprises the sample having and be configured to the biomolecule of described reagent generation chemical reaction.Described method 900 also comprises irradiates described sample area (see 904) by the first and second semiconductor light sources (SLS).Described first and second SLS provide the first and second excitation spectrums respectively.When by described first or the 2nd SLS irradiation, the described biomolecule of described sample can provide the light of instruction association reaction to launch.In addition, described method 900 comprises described light transmitting (see 906) detected from described sample area.Alternatively, described method 900 can comprise and moves described flow cell relative to imaging len and repeat described irradiation and detect operation 904 and 906 (see 908).Figure 60 step that is shown and above-illustrated can repeat, to carry out the repeatedly circulation of sequence measurement.

Figure 39 with Figure 40 illustrates the several functions parts of mobile control system 700 that formed according to an embodiment, that can use together with described imaging system 600.Described mobile control system 700 comprises optical backplane 702 and is coupled to the sample stage 708 of described optical backplane 702 movably.As shown in the figure, described sample stage 702 has supporting surface 704 and bottom surface 705.Described supporting surface and bottom surface 704 and 705 along described Z axis in the face of contrary direction.Described base plate 702 is configured to the major part of the described optics of described optical module 602 (Figure 38) to be supported on described supporting surface 704.Described base plate 702 and described sample stage 708 can be intercoupled movably by intermediate supports 715 and panel 722, so that described sample holder 650 can rotate around described X and Y-axis substantially, be shifted and slide along described X-axis along described Y-axis.

Figure 40 is the isolated skeleton view of the described intermediate supports 715 of described sample stage 708 (Figure 39), electric machine assembly 724 and moveable platform 726.Described electric machine assembly 724 is operatively coupled to described platform 726 and is configured to make described platform 726 along described X-axis Bidirectional slide.As shown in the figure, described intermediate supports 715 comprises tail end 728 and imaging end 730.Described intermediate supports 715 can comprise closes on described imaging end 730 and the pin 746 and 748 given prominence to away from each other along described Y-axis.Close on described imaging end 730, described intermediate supports 715 can comprise lens opening 750, and described lens opening 750 is by allowing described imaging len 623 (Figure 38) size extended through wherein to make and shaping.In the embodiment shown, described pin 746 and 748 has the common line 755 extended through wherein, and described line 755 also extends through described lens opening 750.

Turn back to Figure 39, described platform 726 is coupled to described bottom surface 705 by described intermediate supports 715.Therefore, the weight of described sample stage 708 can be supported by described base plate 702.In addition, described mobile control system 700 can comprise multiple alignment feature 733,735,737 and 739 being configured to locate described sample holder 650.In the embodiment shown, described alignment feature 733,735,737 and 739 is micrometer.Described alignment feature 733 is operably coupled to the described tail end 728 of described intermediate supports 715.When described alignment feature 733 is activated, described tail end 728 can move along the direction of described Z axis.Therefore, described intermediate supports 715 around described pin 746 and 748 (Figure 40), or more specifically, can rotate around described line 755.When described alignment feature 735 and 737 is activated, described sample holder 650 can be shifted along described Y-axis in accordance with the instructions.When described alignment feature 739 is activated, described sample holder 650 can around the turning axle R being parallel to the extension of described X-axis ₇rotate.

Figure 41-42 illustrates skeleton view and the planimetric map of the described optical backplane 702 that can use together with described imaging system 600 (Figure 38) respectively.In some embodiments of described imaging system 600, the one or more of described optics (Figure 38) can have fixing position in described optical module 602, so that described fixing (or static) optics can not move in the operational process of described imaging system 600.Such as, described base plate 702 is configured to the miscellaneous part supporting multiple optics and described imaging system 600.As shown in the figure, described base plate 702 constitutes the structure of the integration substantially had towards the supporting surface (or surface) 704 along described Z-direction.In the embodiment shown, described supporting surface 704 is not continuous and derivable, but can have and be set to the various platform 716-718 of optical module 602 described in predetermined deployment arrangements, pit (or spatial accommodation) 719-721 and parts spatial accommodation 711-714.As shown in figure 42, each of described spatial accommodation 711-714 has respective reference field 781-784.In some embodiments, described reference field 781-784 can be convenient to located by corresponding optics and remain on desired position.

Figure 43 and Figure 44 illustrates front perspective view and the rear portion broken-open perspective view of optical device 732 respectively.As shown in figure 43, described optical device 732 is located relative to orthogonal axle 791-793.Described axle 791 can as above set forth and extend along terrestrial gravitation direction and/or Z axis as described in being parallel to.In a particular embodiment, described optical device 732 is configured to be arranged in the described parts spatial accommodation 713 (Figure 43) of described base plate 702 (part for described base plate 702 is only shown at Figure 43 and Figure 44).

Described parts spatial accommodation 713 has one or more surfaces that restriction wherein can hold the reachable tree region of optics.These one or more surfaces can comprise reference field as described below.In the embodiment shown, described parts spatial accommodation 713 is parts chambeies of the described base plate 701 in described base plate 702 internal depth extension.But described base plate 702 otherwise can form described parts spatial accommodation.Such as, can form the similar fashion in chamber with described base plate 702, described base plate 702 also can have the platform of one or more projection, and the platform of described projection comprises around and limits the surface of described parts spatial accommodation.Therefore, described base plate 702 is plastic partly or ad hoc to provide described parts spatial accommodation.Described base plate 702 can comprise described reference field.In substituting embodiment, sidewall can be installed on described base plate 702 and to be configured to limit described area of space.In addition, other optical device being mounted to described base plate 702 can limit described parts spatial accommodation.As used herein, when element " restriction " parts spatial accommodation, it can be maybe partly limit described parts spatial accommodation that described element ad hoc can limit described parts spatial accommodation.

Described optical device 732 can be removably mounted to the described base plate 702 in described parts spatial accommodation 713, but can be configured to keep fixing position in the operation of described imaging system.But in substituting embodiment, described optical device 732 is not arranged on the inside of described parts spatial accommodation 713, but other places can be arranged on, such as, on the platform of described supporting surface 704.In the embodiment shown, described optical device 732 comprise installing device 734 and be configured to reflect and/or transmission ray through optics 736 wherein.Described installing device 734 is configured to be convenient to support described optics 736 by required orientation and described optics 736 is mounted to described base plate 702.Described installing device 734 comprises component holder 738 and is operably coupled to the biasing element 740 of described retainer 738.

In the embodiment shown, described optics 736 comprises transmitting optical signal through the light filter being wherein filtered into predetermined spectrum simultaneously.But, in substituting embodiment, other optics can be used, such as lens or mirror.As shown in the figure, described optics 736 can comprise face in the opposite direction and limit the thickness T of described optics 736 betwixt ₃optical surface 742 and 744.As shown in the figure, described optical surface 742 and 744 can be the plane being parallel to the continuous and derivable extended each other, makes described thickness T ₃substantial uniform.But in substituting embodiment, described optical surface 742 and 744 can have other profiles.Described optics 736 can have the edge-of-part 751-754 (Figure 43) of multiple restriction periphery or periphery.Described peripheral ring is around described optical surface 742 and 744.As shown in the figure, described periphery is rectangle substantially, but in substituting embodiment, can use other geometric configuratioies (such as circular).

Described retainer 738 is convenient to support described optics 736 by required orientation.In the embodiment shown, described retainer 738 is configured to engage described optical surface 742 and around the extension at least partially of described periphery, to keep described optics 736.Such as, described retainer 738 can comprise the frame extension part 758 that wall portion 756 and the described periphery from described wall portion 756 along described optics 736 (such as described edge-of-part 752 (Figure 43)) extend.In the embodiment shown, described frame extension part 758 can form the bracket of restriction described optics 736 movement.More specifically, described frame extension part 758 can comprise proximal arm 760 and distal arm 762.Described proximal arm 760 extends from described wall portion 756 along described edge-of-part 752 and described axle 791.Described distal arm 762 extends from described proximal arm 760 along described edge-of-part 751.Described distal arm 762 comprises and to extend towards described optics 736 and to engage outshot or the functional part 764 of described optics 736.Still as shown in the figure, described retainer 738 can comprise the clamping components 766 being placed in described frame extension part 758 opposite.Described clamping components 766 and described frame extension part 758 can coordinate to limit described optics 736 and move along described axle 793.Described retainer 738 can clamp a part for the described periphery of described optics 736.

As shown in Figure 43 and 44, described wall portion 756 is configured to engage described optical surface 742.Such as, described wall portion 756 has the matching surface 770 (Figure 43) towards described optics 736.In some embodiments, described wall portion 756 comprises the multiple locating features 771-773 (Figure 43) along described matching surface 770.Described locating features 771-773 is configured to the described optical surface 742 directly engaging described optics 736.When described locating features 771-773 directly engages described optical surface 742, described optical surface 742 (thus described optics 736) is arranged relative to described retainer 738 by required orientation.As shown in figure 43, the described reference field 783 of described parts spatial accommodation 713 also comprises multiple locating features 761-763.Described locating features 761-763 is configured to directly engage described optical surface 744.In addition, described locating features 761-763 can be arranged so that usually, each and described locating features 771-773 of described locating features 761-763 corresponding one relatively.

Still as shown in figure 44, described wall portion 756 has non-matching surface 774, and described non-matching surface 774 is towards the contrary direction relative to described matching surface 770 (Figure 43).Described wall portion 756 comprises the element outshot 776 extended away from described non-matching surface 774 and described optics 736.Described biasing element 740 is configured to be coupled to described element outshot 776.In the embodiment shown, described element outshot 776 and described biasing element 740 extend into the groove 778 of described parts spatial accommodation 713.Described groove 778 makes by the size holding described biasing element 740 and shaping.Described groove 778 has the element surface 780 engaging described biasing element 740.

Figure 45 illustrates the isolated front view of described optical device 732, and Figure 46 illustrates

How described optical device 732 can be removably mounted to described base plate 702.Described optics 736 will be removably installed, described optics 736 can be placed in the parts spatial accommodation 789 of described installing device 734, and described parts spatial accommodation 789 is generally limited by described wall portion 756 (Figure 46), described frame extension part 758 and described clamping components 766.In a particular embodiment, when described optics 736 is placed in described installing device 734, described optics 736 is freely contained in described parts spatial accommodation 789.Such as, described optics 736 can not form interference engagement with described retainer 738.On the contrary, in install operation process, described optics 736 can pass through described wall portion 756, described frame extension part 758, described clamping components 766 and, such as, staff is housed inside in described parts spatial accommodation 789.But in substituting embodiment, described optics 736 can be formed in the space that interference engagement maybe can be limited in only being limited by described retainer 738 with described retainer 738.

About Figure 46, in described install operation process, first described biasing element 740 can be compressed, so as described installing device 734 by and be inserted into described parts receive space 713.Such as, described biasing element 740 can be compressed by the finger of people, to reduce the size of described optical device 732, or described biasing element 740 can by first described biasing element 740 being pressed onto on described element surface 780, then move forward described retainer 738 and enter described parts spatial accommodation 713 to be compressed.Receive in space 713 once described optical device 732 is placed in described parts, described retainer 738 and described optics 736 can move towards described reference field 783, until described optical surface 744 directly engages described reference field 783 by the mechanical energy stored in the biasing element 740 of described compression.More specifically, described optical surface 744 directly can engage the described locating features 761-763 (Figure 43) of described reference field 783.As shown in figure 46, when installing described optics 736, due to described locating features 771-773 (Figure 43), little clearance G can be there is between described optical surface 742 and described matching surface 770 (Figure 43) ₁, and due to described locating features 761-763 (Figure 43), little clearance G can be there is between described optical surface 774 and described reference field 783 ₂.

In described installation site, described biasing element 740 provides the aligning power F be pressed onto by described optical surface 744 on described reference field 783 _a.Described optical surface and reference field 744 and 783 can be configured to arrange described optics 736 by predetermined orientation.Described aligning power F _abe enough to keep described optics 736 by predetermined orientation in the whole operation of described imaging system.In other words, described installing device 734 and described reference field 783 can stop described optics 736 to move along the direction of described axle 792.In addition, in described installation site, described outshot 764 (Figure 43) can be pressed on described edge-of-part 751 (Figure 43), to stop described optics 736 to move along the direction of described axle 791.Described frame extension part 758 and described clamping components 766 can stop or limit described optics 736 to move along the direction of described axle 793.Therefore, described parts spatial accommodation 713 and described installing device 734 can relative to each other configure, to support described optics 736 by predetermined orientation during imaging session.

As shown in figure 45, when described optics 736 is in described installation site, the space segment 798 of described optical surface 744 can in the face of and engage described reference field 783, and the path sections 799 of described optical surface 744 can extend beyond described supporting surface 704 and enters the light path that light signal gets.Still as shown in figure 46, described parts spatial accommodation 713 can extend degree of depth D from described supporting surface 704 _center described base plate 702.

Described biasing element 740 can comprise any can store mechanical can to provide described aligning power F _aelastic component.In the embodiment shown, described elastic component comprises volute spring, then promotes described optical surface 744 against described reference field 783 when described volute spring is compressed.But in substituting embodiment, described elastic component and described parts spatial accommodation can be configured, when described elastic component is stretched then optical surface described in drawing against described reference field.Such as, volute spring can have relative two ends, and wherein one end is connected to the described element surface that is arranged in the groove extended from described reference field and the other end is connected to described retainer.When described volute spring is stretched, described volute spring can provide optics described in drawing against the aligning power of described reference field.In this substituting embodiment, also rubber can be used.

In substituting embodiment, described optics 736 can be labelled to described base plate 702 in order to use bonding agent by described installing device 734.More specifically, described optics 736 can bear against described reference field 783 by described installing device 734.Bonding agent can be deposited the described clearance G between described optical surface 744 and described reference field 783 ₂.After described bonding agent solidification, described installing device 734 can be removed, and described optics 736 is still labelled to described reference field 783 by described bonding agent.

Figure 47 is the block diagram of the described method 800 that assembling optical system is described.Described method 800 comprises provides the optical backplane with parts spatial accommodation (see 802).Described base plate and described parts spatial accommodation can be similar with parts spatial accommodation 713 to base plate 702 mentioned above.Described method 800 also comprises inserts described parts spatial accommodation (see 804) by optics.Described optics can with optics 736 mentioned above similar and comprise be configured to reflect or transmission ray through optical surface wherein.Described optical surface can have towards the reference field of described parts spatial accommodation space segment and extend beyond the path sections that described supporting surface enters light path.Described method 800 also comprises provides the described optical surface of maintenance against described reference field to locate the aligning power (see 806) of described optics.Described optical surface and reference field can be configured to, when providing described aligning power, keep described optics by predetermined orientation.In some embodiments, described method 800 also can comprise and removes described optics (see 808) and alternatively, different opticses is inserted described parts spatial accommodation (see 810).Described different optics can have identical or different optical property.In other words, described different optics can be the refill-unit with identical optical performance, or described different optics can have different optical properties.

Figure 48 and 49 provides skeleton view and the side view of described light source (or exciting light module) 604 respectively.As used herein, light source module comprise one or more light sources (as laser, arc lamp, light emitting diode, laser diode) of being fixed to module frame and also comprise be fixed to as described in module frame relative to as described in one or more light sources fix and one or more opticses in precalculated position (as lens or filtrator).Described light source module can be configured to and is removably coupled in imaging system, so that user can install or change described light source module relatively rapidly.In a particular embodiment, described light source module 604 forms SLS module 604, and described SLS module 604 comprises described first and second SLS614 and 616.As shown in the figure, described SLS module 604 comprises module frame 660 and module lid 662.Multiple image-forming block can be fixed to position relative to each other fixing in described module frame 660.Such as, described first and second SLS614 and 616, described exciting light filtrator 635 and described lens 624 and 625 can be installed on described module frame 660.In addition, described SLS module 604 can comprise the first and second heating radiators 664 (Figure 48) and 666 being configured to the heat energy transmitted respectively from described first and second SLS614 and 616.

Described SLS module 604 and described module frame 660 can make by certain size and shaping, so that individual can grip described SLS module 660 with its hand and easily operate to be fit into described imaging system 600.On this point, described SLS module 604 has the weight that adult can support.

Described SLS module 604 to be configured to be placed in described module spatial accommodation 719 (Figure 41) and to be removably coupled to described base plate 702 (Figure 41).As shown in the figure, described module frame 660 has multiple, and described face comprises installed surface 670 and the field of conjugate action 671 (Figure 48).In the embodiment shown, described module frame 660 is rectangle or block shape substantially, but described module frame 660 can have other shapes in substituting embodiment.Described installed surface 670 is configured to the described base plate 702 being mounted to described module spatial accommodation 719.On this point, described module spatial accommodation 719 at least partially can be shaping to hold and to support described SLS module 604.Similar with described parts spatial accommodation 713, described module spatial accommodation 719 can be limited by one or more surface, and described one or more surface provides the reachable tree region that wherein can hold described SLS module 604.Described surface can be the surface of described base plate 702.Such as, in the embodiment shown, described module spatial accommodation 719 is pits of described base plate 702.Described installed surface 670 can have supplies described base plate 702 and or rather, the profile of described module spatial accommodation 719 substantially.Such as, described installed surface 670 can be plane substantially and comprise from wherein outstanding directing pin 672 (Figure 49), and described directing pin 672 to be configured to be inserted in described base plate 702 in the (not shown) of corresponding hole.Described directing pin 672 can be configured to be convenient to the securing member (as screw) of described module frame 660 to described base plate 702 that be removably coupled.In a particular embodiment, described directing pin 672 is inserted into described base plate 702 with non-normal angle.As shown in figure 49, described heating radiator 666 can be coupled to described module frame 660, there is side-play amount 676 from described installed surface 670 to described heating radiator 666.

Described module frame 660 can be included in channel intersection 685 first and second optical channels 682 and 684 intersected with each other.Described SLS614 and 616 can be fixed to described module frame 660 and the fixed position had relative to each other.Described SLS614 and 616 is located, so that light signal is directed substantially pass respective described optical channel 682 and 684 towards described channel intersection 685 along light path.Described light path can be directed toward described exciting light filtrator 635.In the embodiment shown, described light path is perpendicular to one another, until arrive described exciting light filtrator 635.Described exciting light filtrator 635 by locate to reflect described SLS616 generate described light signal at least partially and transmit described SLS614 generate described light signal at least partially.As shown in the figure, described SLS module 604 is left along common path by common module window 674 from each described light signal of described SLS614 and 616 is directed.Described module window 674 is extended by the described field of conjugate action 671.

Figure 50 is the planimetric map of the described SLS module 604 be installed on described base plate 702.In the embodiment shown, described SLS module 604 is configured to lean against on described base plate 702, so that described terrestrial gravitation g is convenient to described SLS module 604 to support thereon.On this point, described SLS module 604 can provide the integrated device being easy to remove from described optical module 600 or be separated.Such as, remove after the shell (not shown) of described Analytical system or reception enter described optical module, described SLS module 604 can be captured by individual and remove or change.When described SLS module 604 is positioned on described base plate 702, the described field of conjugate action 671 can engage optical device 680.Described optical device 680 can be adjacent with described module window 674, so that the described light signal that described SLS module 604 generates is transmitted by described optical device 680.

Although shown embodiment is described to use the SLS module with the first and second SLS, by other modes, exciting light can be guided on described sample.For example, described SLS module 604 can be included in module frame only a SLS and another optics (such as lens or filtrator) of the fixed position had relative to each other.Equally, plural SLS can be used.In a similar fashion, optical module can comprise only a laser instrument or two or more laser instrument.

But embodiment as herein described is not limited only to have modularization activating system, as described in SLS module 604.Such as, described imaging system 600 can use the light source not being mounted to module frame.More specifically, laser instrument can be mounted directly to other parts of described base plate or described imaging system, maybe can be mounted to framework, and described framework is installed in described imaging system conversely.

Turn back to Figure 38, described imaging system 600 can have image focusing system 840, and described image focusing system 840 comprises described object or sample holder 650, optical system 842 and described imaging detector 610.Described optical system 842 is configured to guide light signal (light such as, from the described sample area 608 of described flow cell 606 is launched) from described sample holder 650 to the detector surface 844 of described imaging detector 610.As shown in figure 38, described optical system 842 comprises described optics 623,644,631 and 642.Described optical system 842 can comprise other opticses.In shown configuration, described optical system 842 has the object or sample plane 846 that close on the setting of described sample holder 650 and the plane of delineation 848 closing on the setting of described detector surface 844.Described imaging detector 610 is configured to obtain object or sample image in described detector surface 844.

In some embodiments, described image focusing system 840 is configured to move the described plane of delineation 848 relative to described detecting device 610 and catch test pattern.More specifically, the described plane of delineation 848 can be moved, so that the described plane of delineation 848 to extend in nonparallel mode relative to described detector surface 844 and intersects with described detector surface 844.The position of described point of crossing can be determined by analyzing described test pattern.Then described position can be used to determine the focal power of described imaging system 600.In a particular embodiment, described image focusing system 840 adopts rotatable mirror, and described rotatable mirror is operably coupled to the actuator for mobile described rotatable mirror.But, described image focusing system 840 can move other opticses described light signal being guided to described detector surface 844, or described image focusing system 840 can move described detecting device 610.In either case, the described plane of delineation 848 relatively can be moved relative to described detector surface 844.Such as, described image focusing system 840 can mobile lens.

In a particular embodiment, described imaging detector 610 is configured to utilize rotatable mirror 642 to obtain test pattern, to determine the focal power of described imaging system 600.According to determined focal power, described imaging system 600 can move described sample holder 650, so that described object or sample are placed in described sample plane 846.Such as, described sample holder 650 can be configured to move the predetermined distance of described sample area 608 (as Δ z represents) with z direction.

Figure 51 is the planimetric map that several parts in described image focusing system 840 are shown.As shown in the figure, described image focusing system 840 comprises the rotatable mirror assembly 850 comprising described mirror 642, the installation component 852 be installed on it by described mirror 642 and is configured to described installation component 852 and described mirror 642 around turning axle R ₆the actuator rotated or rotating mechanism 854.Described mirror 642 is configured to the light signal 863 of reflection receivable from described sample area 608 (Figure 38) towards described imaging detector 610 and in described detector surface 844.In the embodiment shown, described mirror 642 directly reflects described light signal 863 (does not namely have the intervention optics of light signal 863 described in reboot) in described detector surface 844.But, in substituting embodiment, the optics that the described light signal 863 of other impact is propagated may be had.

In the embodiment shown, described image focusing system 840 also comprises and is configured to prevent described mirror 642 from rotating over the positive stop 860 and 862 of predetermined rotational positions.Described positive stop 860 and 862 has relative to described axle R ₆fixed position.Described installation component 852 is configured to according to whether just obtaining sample image or test pattern, around described axle R ₆rotate between described positive stop 860 and 862.Therefore, described mirror 642 can rotate between test position (or orientation) and image space (or orientation).Only for example, described mirror 642 can around described axle R ₆the rotation from about 5 ° to about 12 ° between different position of rotation.In a particular embodiment, described mirror 642 can around described axle R ₆rotate about 8 °.

Figure 52 is the skeleton view of described mirror assembly 850.As shown in the figure, described installation component 852 comprises inner frame 864 and support bracket 866.Described inner frame 864 is configured to be coupled to described mirror 642 and be also coupled to described support bracket 866.Described inner frame 864 and described support bracket 866 can each other and interact with multiple holding screw 868, to provide the inching in the orientation to described mirror 642.On this point, described installation component 852 can form universal mirror installation component.Still as shown in the figure, described installation component 852 is coupled to described rotating mechanism 854.In the embodiment shown, described rotating mechanism 854 comprises direct-driving motor.But, various alternative rotating mechanism can be used, as direct current (DC) motor, electromagnetic coil drive, linear actuators, piezoelectric motor etc.Still as in figure 52, described positive stop 860 can have relative to described rotating mechanism 854 and described axle R ₆fixed position.

As discussed above, described rotating mechanism 854 is configured to around described axle R ₆rotate or rotate described mirror 642.As in figure 52, described mirror 642 has along described axle R ₆the geometric center C extended.The described geometric center C of described mirror 642 is relative to described axle R ₆skew.In some embodiments, described rotating mechanism 854 is configured to be moved between described test position and image space by described mirror 642 in less than 500 milliseconds.In a particular embodiment, described rotating mechanism 854 is configured to being less than 250 milliseconds or be less than in 160 milliseconds and moved between described test position and image space by described mirror 642.

Figure 53 is the schematic diagram of the described mirror 642 at described image space.As shown in the figure, to be reflected by described mirror 642 from the described light signal 863 of described sample area 608 (Figure 38) and be directed toward the described detector surface 844 of described imaging detector 610.According to the configuration of described optical system 842 and the z-position of described sample holder 610, described sample area 608 can fully in focus or not fully in focus (i.e. out of focus).Figure 53 illustrates two plane of delineation 848A and 848B.Described plane of delineation 848A is overlapping with described detector surface 844 substantially, and on this point, corresponding sample image has acceptable or enough focal powers.But described plane of delineation 848B and described detector surface 844 are spaced apart.Therefore, the sample image obtained when described plane of delineation 848B and described detector surface 844 are spaced apart may not have enough focal powers.

Figure 54 and Figure 55 illustrates sample image 870 and 872 respectively.When described plane of delineation 848A is overlapping with described detector surface 844, described sample image 870 is images that described imaging detector 610 detects.When described plane of delineation 848B is not overlapping with described detector surface 844, described sample image 872 is images that described imaging detector 610 detects.(described sample image 870 and 872 comprises the DNA bunch providing fluorescent emission when scheduled excitation spectrum excites.) as shown in Figure 54 and 55, described sample image 870 has acceptable focal power, wherein be clearly defined along described in described sample image 870 bunch each, and described sample image 872 does not have each acceptable focal power be clearly defined of wherein said bunch.

Figure 56 is the schematic diagram of the described mirror 642 at described focal position.As shown in the figure, at the described mirror 642 of described focal position by around described axle R ₆rotate θ angle.Equally, to be reflected by described mirror 642 from the described light signal 863 of described sample area 608 (Figure 38) and be directed toward the described detector surface 844 of described imaging detector 610.But the described optical system 842 in Figure 56 is provided so that the described plane of delineation 848 is moved by relative to described detector surface 844.More specifically, the described plane of delineation 848 can't parallel and described detector surface 844 extend, and on the contrary, intersects at crossing on the same level point PI with described detector surface 844.Although described mirror 642 is in described focal position, described imaging system 600 can obtain the test pattern of described sample area 608.As shown by the circuit diagram of figure 56, the focal degree of described sample area 608 during depending on imaging session, described crossing on the same level point PI can betide the diverse location in described detector surface 844.

Such as, Figure 57 and Figure 58 illustrates test pattern 874 and 876 respectively.Described test pattern 874 represents the image obtained when described sample area 608 is in focus, and described test pattern 876 represents the image obtained when described optical system 842 out of focus.As shown in the figure, described test pattern 874 has focal zone or position FL ₁(distance of itself and described reference edge 880 is XD ₁) and described test pattern 876 there is focal zone or position FL ₂(distance of itself and described reference edge 880 is XD ₂).Described focal position FL ₁and FL ₂can be determined by image analysis module 656 (Figure 38).

Confirm the described focal position FL in described test pattern 874 and 876 ₁and FL ₂, described image analysis module 656 can determine the position of best focal power in corresponding test pattern.More specifically, described analysis module 656 can determine the focusing score of the difference of the x-dimension along described test pattern 874 and 876.Described analysis module 656 can calculate the focusing score on each point based on one or more image quality parameter.The example of image quality parameter comprises the square error in picture contrast, spot size, signal noise ratio (snr) of image and described image between pixel.For example, when calculating focusing score, described analysis module 656 can calculate the variation factor of contrast in described image.The variation factor of described contrast represents the variable quantity in the selected part of image or image between image pixel intensities.As a further example, when calculating focusing score, described analysis module 656 can calculate the size of the hot spot deriving from described image.Described hot spot can be expressed as Gauss's hot spot and size can be measured as full width at half maximum (FWHM), and in this case, less spot size is usually relevant to the focusing improved.

After determining focal position FL described in described test pattern, described analysis module 656 then can measure or determine described focal position FL and described reference edge 880 is spaced apart or the described distance XD that separates.Described distance XD then can be relevant relative to the z-position of described sample plane 846 to described sample area 608.Such as, described analysis module 656 can determine the described distance XD shown in Figure 58 ₂correspond to the described sample area 608 with described sample plane 846 distance, delta z.On this point, then described sample holder 650 can be moved described distance, delta z, with described sample area 608 mobile in described sample plane 846.Therefore, the described focal position FL in test pattern can indicate described sample area 608 relative to the position of described sample plane 846.As used herein, phrase " indicates described object (or sample) relative to the position of described object (or sample) plane " and comprises and utilizes factor (such as described focal position), to provide more suitably for determining model or the algorithm of described distance, delta z.

Figure 59 is the block diagram of a kind of method 890 that focus for control both optical imaging system is described.Described method 890 comprise provide there is rotatable mirror and be configured to guide light signal to the optical system (see 892) in detector surface.Described detector surface can be similar to described detector surface 844.Described optical system can have the object plane of closing on object, as described in sample plane 846.Described optical system also can have the plane of delineation closing on described detector surface, as described in the plane of delineation 848.Described rotatable mirror can be rotatable between image space and focal position.

Described method 890 also comprises the described mirror of rotation to described focal position (see 894) and the test pattern (see 896) obtaining described object when described mirror is in described focal position.Described test pattern can have best focal power at focal position.Described focal position can indicate described object relative to the position of described object plane.In addition, described method 890 also can to comprise described object move based on described focal position towards described object plane (see 898).

Be appreciated that description is above to illustrate, and unrestriced.Such as, above-mentioned embodiment (and/or its aspect) can be combined with each other.In addition, many amendments can be made to adapt to particular case or material to embodiment, and not deviate from scope of the present invention.Although concrete parts described herein and process are intended to the parameter limiting various embodiment, they are never restrictive but exemplary embodiments.After checking description above, other embodiments many will be apparent to one skilled in the art.Therefore, the four corner of equivalent that described scope of the present invention should be endowed together with these claims with reference to appended claim is determined.In the appended claims, term " comprises " and " wherein " is used as the simple and easy English equivalent of " comprising " and " wherein " separately.In addition, in claim hereafter, " first ", " second " and " the 3rd " etc. are only with marking, and are not intended to force numerical requirements to its object.In addition, writing of claim restriction is not hereafter with the form of means-plus-function and can't makes explanations based on 35U.S.C. § 112 the 6th section, unless and until the restriction of described claim connects the invalid statement of function of further structure after clearly using phrase " to refer to ".

Claims (5)

1. arrange a method for the fluidic hardware being used for sample analysis, described method comprises:

There is provided tool chlamydate fluidic hardware, described shell comprises spatial accommodation and is placed in the flow cell that can float along the object plane of described spatial accommodation of described spatial accommodation inside; Described shell has the recess that the described spatial accommodation of next-door neighbour is arranged;

Being arranged on by described fluidic hardware has in the supporting construction of alignment members, and described alignment members is inserted into by corresponding recess;

Described flow cell in mobile described spatial accommodation; When described flow cell moves in described spatial accommodation, described alignment members engages the edge of described flow cell and stops described flow cell in the movement of the described object plane of described spatial accommodation.

2. method according to claim 1, the described flow cell of wherein said movement comprises startup registration arm to be pressed onto in described alignment members by described flow cell.

3. method according to claim 1, wherein said fluidic hardware comprises the lid component being rotatably coupled to described shell, and described lid component comprises packing ring, entrance and exit passage; Wherein said lid component is rotatable around the turning axle between installation site and disengaged position; When described packing ring is in described installation site, the entrance and exit rough alignment of described entrance and exit passage and described flow cell.

4. method according to claim 3, wherein when the entrance and exit of flow cell described in described entrance and exit passage rough alignment, described packing ring is pressed on the edge of described flow cell; Described packing ring limits the movement of described flow cell in described spatial accommodation.

5. method according to claim 1, wherein said flow cell has towards rightabout first and second faces, pond, described first and second faces, pond extend along described object plane, and described method comprises on one that lid component is pressed in described first and second faces, pond further with the movement of restriction perpendicular to the direction of described object plane.