WO2022125070A1

WO2022125070A1 - Sample preparation with plunge and mixing chambers

Info

Publication number: WO2022125070A1
Application number: PCT/US2020/063733
Authority: WO
Inventors: Quinton Buford WEAVER; Si-lam Julius CHOY; Seth Stephen HADDIX; Kelly B. Smith; John Michael LAHMANN
Original assignee: Hp Health Solutions Inc.
Priority date: 2020-12-08
Filing date: 2020-12-08
Publication date: 2022-06-16

Abstract

In one example in accordance with the present disclosure, A sample preparation device is described. The sample preparation device includes a mixing chamber to receive a sample and a mixer disposed within the mixing chamber to mix the sample. The sample preparation device also includes a actuator to transmit a torque to the mixer. A seal separates the mixing chamber from a downstream component and a plunger is disposed within the mixing chamber to direct contents of the mixing chamber to the downstream component.

Description

SAMPLE PREPARATION WITH PLUNGE AND MIXING CHAMBERS

BACKGROUND

[0001] Analytic chemistry is a field of chemistry that uses instruments to separate, identify, quantify, and study matter. Biochemistry is a field of chemistry that includes the study and analysis of the chemistry of living organisms such as cells. Cell lysis is a process of rupturing the cell membrane to extract intracellular components for purposes such as purifying the components, retrieving deoxyribonucleic acid (DNA), ribonucleic acid (RNA), proteins, polypeptides, metabolites, or other small molecules contained therein, and analyzing the components for genetic and/or disease characteristics. Cell lysis bursts a cell membrane and frees the inner components. The fluid resulting from the bursting of the cell is referred to as lysate.

BRIEF DESCRIPTION OF THE DRAWINGS

[0002] The accompanying drawings illustrate various examples of the principles described herein and are part of the specification. The illustrated examples are given merely for illustration, and do not limit the scope of the claims.

[0003] Fig. 1 is a block diagram of a sample preparation device with a plunger and mixing chamber, according to an example of the principles described herein.

[0004] Fig. 2 is a block diagram of a sample preparation device with a plunger and mixing chamber, according to an example of the principles described herein. [0005] Fig. 3 is an isometric cross-sectional view of a cartridge with multiple sample preparation devices, according to an example of the principles described herein.

[0006] Fig. 4 is an exploded view of the components of the plunger and mixing chamber, according to an example of the principles described herein. [0007] Figs. 5A - 5D are views of the actuator to transmit a torque from a motor to the mixer, according to an example of the principles described herein. [0008] Fig. 6 is a view of a portion of the mixer shaft that engages with the actuator, according to an example of the principles described herein.

[0009] Figs. 7A - 7C depict interaction of a driving shaft with the sample preparation device, according to an example of the principles described herein. [0010] Figs. 8A - 8C depict the translation of the plunger to direct contents of the mixing chamber to a downstream component, according to an example of the principles described herein.

[0011] Fig. 9 is a cross-sectional view of the sample preparation device plunger and mixing chamber, according to another example of the principles described herein.

[0012] Fig. 10 is a cross-sectional view of the sample preparation device plunger and mixing chamber, according to another example of the principles described herein.

[0013] Fig. 11 is a cross-sectional view of the seal of the sample preparation device plunger and mixing chamber, according to an example of the principles described herein.

[0014] Fig. 12 is a cross-sectional view of the seal of the sample preparation device plunger and mixing chamber, according to an example of the principles described herein.

[0015] Fig. 13 is a cross-sectional view of the seal of the sample preparation device plunger and mixing chamber, according to an example of the principles described herein.

[0016] Fig. 14 is a cross-sectional view of the seal of the sample preparation device plunger and mixing chamber, according to an example of the principles described herein. [0017] Fig. 15 is a cross-sectional view of the seal of the sample preparation device plunger and mixing chamber, according to an example of the principles described herein.

[0018] Figs. 16A and 16B are cross-sectional views of the sample preparation device plunger and mixing chamber, according to another example of the principles described herein.

[0019] Fig. 17 is a flow chart of a method of sample preparation, according to an example of the principles described herein.

[0020] Fig. 18 depicts an interaction of a driving shaft with the sample preparation device, according to an example of the principles described herein. [0021] Throughout the drawings, identical reference numbers designate similar, but not necessarily identical, elements. The figures are not necessarily to scale, and the size of some parts may be exaggerated to more clearly illustrate the example shown. Moreover, the drawings provide examples and/or implementations consistent with the description; however, the description is not limited to the examples and/or implementations provided in the drawings.

DETAILED DESCRIPTION

[0022] Cellular analytics is a field of chemistry that uses instruments to separate, identify, quantify, and study cells and their internal components matter. A wealth of information can be collected from a cellular sample. For example, a study of a patient’s cell may lead to diagnosis of diseases of a patient. As a particular example, cellular analysis may be used to detect viral nucleic acid, for example in a flu test. Moreover, study of cells may lead to the development of medications to treat certain diseases and disorders.

[0023] The intracellular components of the cell also provide valuable information about a cell. Cell lysis is a process of extracting intracellular components from a cell and can also provide valuable information about a cell. During lysis, the intracellular components are extracted for purposes such as purifying the components, retrieving DNA and RNA, proteins, polypeptides, metabolites, and small molecules or other components therein, and analyzing the components for genetic and/or disease characteristics. Cell lysis ruptures a cell membrane and frees the inner components. The fluid containing the inner components is referred to as lysate. The contents of the cell can then be analyzed by a downstream system. Prior to analysis, the sample to be analyzed is prepared. During preparation, the sample may be lysed and components of the lysate may be bound to magnetic microparticles. Other operations may also be carried out to prepare the same for cellular analysis.

[0024] While cellular analytics is useful in cellular analysis, refinements to the operation may yield more detailed analysis results. For example, lysing and binding may be carried out by heating, cooling, and mixing the sample. In general, it may be difficult to heat and mix a sample. Furthermore, the manual execution of these operations may increase the time to perform a complete analysis of the sample.

[0025] Accordingly, the present specification describes a system that automates the steps of heating and mixing a sample. For example, components of a lysate sample may be bound to magnetic microparticles which are particles that may be surface-activated to selectively bind with a biological component or that may be bound to a biological component from a biological sample. As they are magnetic, the microparticles and the bound biological component may be moved about throughout the remaining preparation and analysis operations via an external magnet. In some examples, the magnetic microparticles are paramagnetic microparticles; in some other examples, the magnetic microparticles are paramagnetic beads.

[0026] That is, in biological assays, a biological component can be intermixed with other components in a biological sample that can interfere with subsequent analysis. As used herein, the term “biological component” may refer to materials of various types, including proteins, cells, cell nuclei, nucleic acids, bacteria, viruses, or the like, that can be present in a biological sample. A “biological sample” may refer to a fluid or a dried or lyophilized material obtained for analysis from a living or deceased organism. Isolating the biological component from other components of the biological sample may permit subsequent analysis without interference and may increase an accuracy of the subsequent analysis. In addition, isolating a biological component from other components in a biological sample may permit analysis of the biological component that would not be possible if the biological component remained in the biological sample. In some examples, the biological component of interest that will be bound to the magnetic microparticles are nucleic acids (such as DNA and RNA)

[0027] The sample and/or magnetic microparticles are then automatically directed into a fluid isolation chamber where the same may be further operated upon. For example, in the fluid isolation chamber, the lysate may be combined with at least one reagent. The present specification describes preparing multiple samples in parallel in different sample preparation devices.

[0028] In general, the present specification describes a sample preparation device. The sample preparation device includes a mixing chamber with a mixer, plunger, seal, and actuator. The sample preparation device mixes and heats the sample inside the mixing chamber. The heating may be done by contacting the outside of the sample preparation device with a heat block and binding may be done by mixing the sample and magnetic microparticles using the mixer.

The mixer of the present specification may accelerate the heating by 1) increasing the convection rate of the fluid and 2) increasing the heatable surface area of the fluid in direct contact with the heated walls. Once the component of the lysate is bound to the magnetic microparticles, the lysate is transferred out of the mixing chamber into a density gradient column, i.e., the fluid isolation chamber, for further processing.

[0029] Such a device also maintains a liquid-sealed volume during mixing/heating for reduced contamination risk during sample preparation. Following heating/mixing, the mixing chamber may be opened and the contents delivered to a fluid isolation chamber via a physical probe or air pressure.

[0030] As such, the sample preparation device, when coupled with a host station which controls the different components (i.e., actuator, mixer, plunger), delivers a ready to analyze sample without additional user interaction beyond sample input and removal. In some examples, multiple sample preparation devices may be inserted into a cartridge such that multiple samples may be prepared in parallel. Thus, the sample preparation device provides a high speed, high volume throughput which enables testing and analysis capacity beyond what a single user could prepare manually.

[0031] The present specification focuses on the mixing chamber and components therein that enable the host station, that is the device into which the sample preparation device is inserted, to mix the fluid inside the mixing chamber and to deliver that fluid from the mixing chamber to the fluid isolation chamber. That is, these components within the mixing chamber mix the sample, open/unseal the mixing chamber, and deliver the sample to the fluid isolation chamber. In some examples, the mixing chamber may be opened using a physical probe (piercing) or by using air pressure. Either unsealing mechanism is facilitated by the downward motion of the plunger in the mixing chamber. Such a mixing chamber provides a system that lyses, binds, cools, and delivers the fluid and is housed in a single mixing chamber.

[0032] In an example, the direction of rotation of the mixer may change based on a function. In a first direction, the mixer may be rotated to pierce the seal between the mixing chamber and the fluid isolation chamber. When rotated in another direction, the plunger may be advanced to push fluid out of the mixing chamber. The seal of the present sample preparation device dispenses fluid and reduces fluid disturbances (e.g. spray). Moreover, the sample remains vented to atmosphere until plunged. Once plunged, the remaining air in the mixing chamber may become a fluid driving force. The mixing chamber is sealed during plunging and remains sealed after use for disposal. Such lysing, mixing, and delivery may be accomplished with rotation and longitudinal translation of the plunger and/or mixer.

[0033] Specifically, the present specification describes a sample preparation device. The sample preparation device includes a mixing chamber to receive a sample and a mixer disposed within the mixing chamber to mix the sample. A actuator transmits a torque to the mixer. A seal separates the mixing chamber from a downstream component and a plunger disposed within the mixing chamber opens the seal and direct contents of the mixing chamber to the downstream component. [0034] In an example, a mixer shaft aligns longitudinally with the mixing chamber and passes through the plunger. The mixer shaft may be attached to the actuator. In this example, the actuator includes a protrusion to interface with a notch in a driving shaft of a host station such that as the driving shaft rotates in a first direction, the actuator and mixer rotate in the first direction. When the protrusion aligns with the notch and responsive to a translational force against the actuator, the actuator is to move the mixer to open the seal. Responsive to the driving shaft being rotated in a second direction that is opposite the first direction, the protrusion aligns with a slot in the driving shaft such that the actuator disengages from the driving shaft. When the protrusion aligns with the slot and responsive to a translational force, the plunger translates to dispel contents of the mixing chamber.

[0035] In an example, when in an evacuated state, the plunger is to block a port to the mixing chamber. In an example, the seal is selected from the group consisting of a film, a ball, a plug, a septum, a pipette tip to be acted upon by the mixer to allow contents of the mixing chamber to pass, and a sliding seal to be translated to expose a bypass channel to allow contents of the mixing chamber to pass. In an example, the seal is ruptured by a needle affixed to a bottom of the mixing chamber.

[0036] The present specification also describes a method. According to the method, a sample is lysed by rotating a mixer of a sample preparation device disposed within a host station. Once mixed, a seal between a mixing chamber of the sample preparation device and a fluid isolation chamber of the sample preparation device is ruptured. The contents of the mixing chamber are driven towards the fluid isolation chamber by translating a plunger through the mixing chamber. In an example, the seal is ruptured via a mechanical interaction or via air pressure.

[0037] In another example, a sample preparation device includes a mixing chamber to receive a sample, an input port to introduce the sample into the mixing chamber, and a mixer disposed within the mixing chamber to mix the sample. The sample preparation device also includes a actuator to transmit a torque to the mixer, a film seal to separate the mixing chamber from a downstream component, and a plunger disposed within the mixing chamber to direct contents of the mixing chamber to the downstream component. In this example, the sample preparation device includes a fluid isolation chamber to receive the contents and an output to dispense contents of the fluid isolation chamber.

[0038] In an example, the sample preparation device is disposed in a cartridge along with other sample preparation devices.

[0039] In an example, the plunger, actuator, and mixer are a single integrated component. In another example, the plunger and actuator are a single integrated component and a plunger shaft is to slide through a mixer opening to rupture the film seal.

[0040] In summary, using such a sample preparation device 1 ) enables automated lysing of cells and binding components such as nucleic acids to magnetic microparticles; 2) maintains a liquid sealed chamber during mixing/heating for minimal contamination risk during sample preparation where air can escape but not liquid; 3) allows for controlled delivery of the lysate to the fluid isolation chamber; 4) enables multiple instances of the sample preparation to occur in the same physical space; 5) executes one sample transfer operation; 6) reduces stranded fluid; and 7) remains sealed for disposal. However, the devices disclosed herein may address other matters and deficiencies in a number of technical areas.

[0041] Turning now to the figures, Fig. 1 is a block diagram of a sample preparation device (100) with a plunger (110) and mixing chamber (102), according to an example of the principles described herein. As described above a sample may be prepared before it is analyzed or processed. Such preparation may include lysing a sample and binding a specific element in the lysate, such as for example nucleic acid, to magnetic microparticles. The sample preparation device (100) of the present specification carries out at least a portion of this sample preparation. Specifically, the sample preparation device (100) includes a mixing chamber (102), which is a volume, to receive the sample. In some examples, the sample may be a biological sample, that is the sample may include biological components to be studied and analyzed. Within the mixing chamber (102), operations such as lysing and binding a specific element present in the lysate, such as for example nucleic acid, to magnetic microparticles are performed. In general, lysis refers to the agitation of a cell with the objective of rupturing a cell membrane. Lysis ruptures a cellular particle membrane and frees the inner components. The fluid containing the inner components is referred to as lysate. The contents of the cellular particle can then be analyzed by a downstream system.

[0042] The sample cells may be lysed by heating the walls of the mixing chamber (102) which causes the cells to rupture. In some examples, the mixing chamber (102) may also include a chemical compound that lyses the cells. That is, in addition to being a volume wherein the sample is mixed with a reagent, the mixing chamber (102) may store the reagent to be used. Accordingly, during manufacture, a reagent may be deposited in the mixing chamber (102) rather than being input as part of the sample preparation operation. The mixer (104) disposed within the mixing chamber (102) may stir the contents of the mixing chamber to aid and expedite the lysing operation.

[0043] Once lysed, a component of the lysate, for example nucleic acid, may be bound to magnetic microparticles that are disposed within the mixing chamber (102). In this example, a pellet that contains the magnetic microparticles may be disposed within the mixing chamber (102). When introduced, the sample may dissolve the structure of the pellet to release the magnetic microparticles contained therein. The action of the mixer (104) also places the contents of the lysate and the magnetic microparticles in proximity to one another such that they may bind together. For example, the magnetic microparticles may be heavier than the solution in the mixing chamber (102) and may otherwise settle to the bottom of the mixing chamber (102). The mixer (104) introduces turbulence that distributes the magnetic microparticles more uniformly throughout the matrix of the solution such that there are more opportunities for the magnetic microparticles to bind with a specific component of the lysate.

[0044] The mixer (104) may include a mixing head, blade, or paddle on the end of the shaft. In one example, the mixer (104) may be rotated manually. In another example, the mixer (104) may be rotated by a motor in the device in which the sample preparation device (100) is installed. That is, the sample preparation device (100) may be installed in a host station. The host station may include a motor and a shaft that interacts with the mixer (104) to rotate the mixer (104) to agitate the contents in the mixing chamber (102).

[0045] In this example, the sample preparation device (100) also includes an actuator (106) to transmit a torque from the motor of the host station to the mixer (104). That is, the actuator (106) provides an interface through which this motion is imparted to the mixer (104) and the plunger (110). On one end, the actuator (106) may include interface surfaces to interact with a driving shaft of the host station and therefore to be driven by the driving shaft. The other end of the actuator (106) may include attachment surfaces that couple the actuator (106) to the mixer (104) and that drive the mixer (104). Figs. 5A - 5D below depict example views of the actuator (106).

[0046] The sample preparation device (100) also includes a seal (108) to separate the mixing chamber (102) from a downstream component. In an example, the downstream component is a fluid isolation chamber where additional operations are performed. For example, in the fluid isolation chamber, the lysate and magnetic microparticles may be combined with a reagent. As another example, the sample may be concentrated and purified within the fluid isolation chamber. For example, the sample may be purified by a density gradient within the fluid isolation chamber and magnetic motion imparted within the fluid isolation chamber. Furthermore, coupled to the fluid isolation chamber may be liquid reagents such as master mix to further process the sample.

[0047] A specific description of example operations carried out in the fluid isolation chamber are now provided. Initially, a fluid stack of washing fluid may be introduced into the fluid isolation chamber. The lysate may then be delivered from the mixing chamber (102) on top of the fluid stack. Magnets of the host station may be used to aggregate the magnetic microparticles in the lysate and the clump may be swept out of the lysate, down the fluid isolation chamber, and into an output. In some examples, reaction chemicals such as master mix, are inserted into the fluid isolation chamber. The lysate, magnetic microparticles, and/or chemicals are dispensed by the output into a receptacle for analysis. [0048] Prior to lysing and binding, it may be desirable to seal the mixing chamber (102) to prevent flow out of the mixing chamber (102). Accordingly, the mixing chamber (102) includes a seal (108). Upon rupture of the seal (108), the contents flow to the fluid isolation chamber for further preparation.

Examples of different seals (108) are provided below in connection with Figs. 8A - 16.

[0049] The sample preparation device (100) also includes a plunger (110) disposed within the mixing chamber (102). The plunger (110) operates to open the seal (108) and direct the contents of the mixing chamber (102) to the downstream component. That is, once mixed, the seal (108) may be opened. The force of gravity may draw the fluid through the fluid isolation chamber. This motion may be accelerated by the plunger (110) which is activated to push the contents out. As described above, the motion of the plunger (110) may be controlled by the motor and the actuator (106). Figs. 8A - 8C depict the operation of motor of the host station and the actuator (106) to move the plunger (110) to expel the contents of the mixing chamber (102).

[0050] Fig. 2 is a block diagram of a sample preparation device (100) with a plunger (110) and mixing chamber (102), according to an example of the principles described herein. As in the previous example, the sample preparation device (100) may include a mixing chamber (102), mixer (104), actuator (106), seal (108), and plunger (110). In this example, the seal may be a film seal (108) that is ruptured either by the mixer (104) blade or by air pressure as the plunger (110) is driven.

[0051] In this example, the sample preparation device (100) also includes an input port (212) to receive the sample. That is, the input port (212) may be in fluid communication with the mixing chamber (102) such that a user may introduce the sample into the mixing chamber (102). For example, a user may insert a pipette into the input port (212) and expel the contents therein into the mixing chamber (102) to begin the sample preparation operation. [0052] The sample preparation device (100) may also include a fluid isolation chamber (214) to receive the contents of the mixing chamber (102). The fluid isolation chamber (214) provides a fluid path for collecting the magnetic microparticles (using external magnets) out of the lysate, pulling the magnetic microparticles towards the output (216), and eventually dispensing them into the waiting receptacle for analysis. The magnetic microparticles may also be cleaned in the fluid isolation chamber (214). In some examples, within the fluid isolation chamber (214), the lysate may be mixed with a reagent.

[0053] The sample preparation device (100) may also include an output (216) to dispense the contents of the fluid isolation chamber (214). That is, as described above, the sample preparation device (100) may prepare the sample for analysis. The output (216) of the sample preparation device (100) may eject the prepared sample onto a surface such that the analysis may be performed. In an example, the surface may be a well plate with individual wells.

[0054] Fig. 3 is an isometric cross-sectional view of a cartridge (318) with multiple sample preparation devices (Fig. 1 , 100), according to an example of the principles described herein. As described above, in some examples, the host station may operate to prepare multiple samples in parallel. For example, as described above, each sample preparation device (100) analyzes a single sample. Accordingly, multiple parallel sample preparation devices (100) allow multiple samples to be analyzed at the same time, rather than analyzing a single sample at a time.

[0055] Accordingly, the sample preparation devices (Fig. 1 , 100) may be disposed in a cartridge (318) along with other sample preparation devices (Fig.

1 , 100). The cartridge (318) is insertable into host station, which host station provides the signals and mechanical forces to 1) activate the mixer (104), lyse the sample, rupture the seal (110), and eject the prepared sample on to a surface.

[0056] Fig. 3 clearly depicts the mixing chamber (102) with its associated mixer (104), actuator (106), plunger (108), seal (110), and input port (212). Fig. 3 also clearly depicts the fluid isolation chamber (214) and the output (216). [0057] The output (216) may include an air blister. As the air blister is depressed, pressure forces the fluid out the sample preparation device (Fig. 1 , 100) and onto the surface, such as a well plate

[0058] Specific examples of the operation of the sample preparation device (Fig. 1 , 100) are now provided. As a general example, a biological sample may be eluted into a transport medium. The biological sample may include a biological component of interest, such as for example nucleic acids. The biological sample may then be prepared, for example by lysing the cell which contains the biological component of interest, such as the nucleic acid and binding the nucleic acid to magnetic microparticles. The nucleic acids may then be mixed with a master mix. At this stage, the prepared sample may be ejected onto a surface, such as a titration plate where the samples may be further processed, for example by performing PCR analyses in cases where magnetic microparticles are bound to nucleic acids.

[0059] As a more specific example, a swab with a sample may be inserted into a transport vial where it is eluted into a medium. A portion of the sample is introduced, for example via a pipette, into the mixing chamber (102) via the input port (212). Introduction of the sample dissolves a holding pellet and releases the magnetic microparticles disposed therein. The sample may be sequentially and/or simultaneously heated, via heat blocks heating walls of the mixing chamber (102) and rotation of the mixer (104). The sample is lysed by heating the sample to a temperature of 80 degrees Celsius (° C) which in one example ruptures the membrane walls spilling the lysate. At this point, the lysate may be cooled to a temperature of around 56° C in an example, yet the mixing may continue. A chemical reaction binds the sample a component of the lysate, such as a nucleic acid, to the magnetic microparticles. Increased binding is provided via action of the mixer (104) to agitate the sample to promote interaction between the sample lysate and the magnetic microparticles.

[0060] At some point prior to rupturing of the seal (110), a wash buffer may be introduced into the fluid isolation chamber (214). A wash buffer refers to a composition that may wash the magnetic microparticles of impurities that may be in the sample and that may inhibit downstream processes such as PCR. The wash buffer also forms a continual fluid path from the lysate to the output (216). Such a wash buffer may rinse the nucleic acid/magnetic microparticles removing a reagent and preparing the sample for application of another reagent. In an example, the wash buffer may include water, some salts to control pH, other salts to help keep the nucleic acids stay bound to the magnetic microparticles, a surfactant to help keep the magnetic microparticles distributed, and preservatives/biocides. In some examples, the wash buffer may include a densifier such as iodixanol to create the density gradient-based purification method in the fluid isolation chamber (214). In other examples, the wash buffer may include alcohol (ethanol or isopropanol), oils, other surfactants, etc.

[0061] The lysate may then be introduced into the fluid isolation chamber (214) via 1) action of the mixer (104) to rupture the seal (108) and 2) action of the plunger (110) to drive the fluid. As described above, this motion may be driven by the motor and transmitted to the mixer (104) and the plunger (110) via the actuator (106).

[0062] When in the fluid isolation chamber (214), certain operations may be performed to further process the sample. Once prepared, the sample may be ejected via the output (216) to be subsequently analyzed.

[0063] Fig. 4 is an exploded view of the components of the plunger (110) and mixing chamber (Fig. 1 , 102), according to an example of the principles described herein. Specifically, Fig. 4 depicts the mixer (104), plunger (110), and actuator (106). As depicted in Fig. 3, the mixer (104) aligns longitudinally with the mixing chamber (Fig. 3, 102). The mixer (104) also passes through the plunger (110). Fig. 4 also depicts additional components of the plunger (110). Specifically, the plunger (110) may include O-ring seals (420-1 , 420-2) that seal the sample within the mixing chamber (102). That is, the O-ring seals (420-1 , 420-2) create a seal that allows the plunger (110) to move to expel the contents of the mixing chamber (Fig. 1 , 102), all while preventing sample fluid from leaking out the sample preparation device (Fig. 1 , 100).

[0064] Figs. 5A - 5D are views of the actuator (106) which transmits a torque from a motor to the mixer (Fig. 1 , 104), according to an example of the principles described herein. As described above, the actuator (106), when driven by a motor of a host station, 1 ) spins the mixer (Fig. 1 , 104) and 2) translates the plunger (Fig. 1 , 110) and mixer (Fig. 1 , 104). Accordingly, the actuator (106) has certain features to allow the rotational and translational force transmission.

[0065] Specifically, the actuator (106) may include protrusions (522-1 , 522-2) that interface with a notch in a driving shaft of a host station. The protrusions (522) engage with the driving shaft to transmit torque. That is, the surfaces of the protrusions (522-1 , 522-2) fit into a notch and are selectively coupled such that as the driving shaft rotates, so does the actuator (106). Accordingly, as the driving shaft rotates in a first direction, the actuator (106) and the mixer (Fig. 1 , 104) rotate in the first direction.

[0066] The protrusions (522) when positioned in the notch, may also transmit a translational force, which translational force moves the mixer (Fig. 1 , 104) to pierce the seal (Fig. 1 , 108). An example of the interaction of the protrusions (522) and the driving shaft to transmit rotational and translational force is provided below in connection with Figs. 7A - 7C. In this example, the actuator (106) may spin freely within the plunger (110).

[0067] The actuator (106) is attached to the mixer (Fig. 1 , 104). Specifically, the actuator (106) may include deflecting members (524-1 , 524-2) that upon insertion of a mixer (Fig. 1 , 104), interact with latches on the mixer (Fig. 1 , 104) to retain the mixer (Fig. 1 , 104) in place. Fig. 6 depicts an example of the mixer (Fig. 1 , 104) with the latches disposed thereon. The actuator (106) may also include flat surfaces (526) that press against corresponding surfaces of the mixer (Fig. 1 , 104) to transmit the torque. That is, the flat surfaces (526) of the actuator (106) press against surfaces of the mixer (Fig. 1 , 104) to rotate the mixer (Fig. 1 , 104).

[0068] Fig. 6 is a view of a portion of the mixer (104) shaft that engages with the actuator (Fig. 1 , 106), according to an example of the principles described herein. As described above, the mixer (104) shaft may include latches (628-1 , 628-2) that interface with the deflecting members (Fig. 5, 524-1 , 524-2) to retain the mixer (104) firmly in contact with the actuator (Fig. 1 , 106). In this example, as the mixer (104) is inserted into the actuator (Fig. 1 , 106), the latches (628-1 , 628-2) deflect the deflecting members (Fig. 5, 524). When the latches (628) clear, the deflecting members (Fig. 5, 524) rebound underneath the latches (628-1 , 628-2) as depicted in Fig. 8A to prevent the mixer (104) from being decoupled from the actuator (Fig. 1 , 106).

[0069] Fig. 6 also depicts the surface (630) that interacts with one of the flat surfaces (Fig. 5, 526) of the actuator (Fig. 1 , 106) to rotate the mixer (104). That is, the flat surface (Fig. 5, 526) of the actuator (Fig. 1 , 106) interfaces with the surface (630) to transmit any rotational force to the mixer (104).

[0070] Figs. 7A - 7C depict interaction of a driving shaft (732) with the sample preparation device (Fig. 1 , 100), according to an example of the principles described herein. As described above, the host station may include a driving shaft (732) that is coupled to a motor which provides rotational movement. The driving shaft (732) includes an L-shaped recess including a notch and a slot. During use, the sample preparation device (Fig. 1 , 100) may be aligned such that the protrusion (522) aligns with the slot. When the sample preparation device (Fig. 1 , 100) is fully seated, that is when an end of the plunger (110) contacts an end surface of the driving shaft (732), the driving shaft (732) may be rotated as indicated by the arrow (734). This causes the protrusion (522) to align with, or sit in, the notch as depicted in Fig. 7A. Due to this interface between the protrusion (522) and the notch of the driving shaft (732), as the driving shaft is rotated in the direction indicated by the arrow (734), the actuator (106) and mixer (104) are also rotated in the direction indicated by the arrow (734). In this example, the actuator (Fig. 1 , 106) may spin freely within the plunger (110). That is, the actuator (Fig. 1 , 106) and mixer (104) may rotate while the plunger (110) is rotationally stationary. Accordingly, the motor, via the 1 ) driving shaft (734) and actuator protrusions (522) and 2) the attachment of the mixer (104) to the actuator (Fig. 1 , 106), facilitate rotation of the mixer (104) to mix the sample during lysing and binding.

[0071] The interface between the notch and the protrusion (522) may also transmit the translational force. That is, when the protrusion (522) aligns with the notch and responsive to a translational force from the driving shaft (732) against the actuator (Fig. 1 , 106) as indicated by the arrow (736), the actuator (Fig. 1 , 106) moves the mixer (104) to open the seal. Figs. 8A - 8C depict examples of the movement of the actuator (Fig. 1 , 106), and mixer (104) to rupture the seal.

[0072] In some examples, the interface between the driving shaft (732) and the actuator protrusions (522) may be light as to not overcome sticking friction on the seal and prematurely plunge. Moreover, the plunger (110) supports the weight of the driving shaft (732) during mixing. In some examples, the driving shaft (732) is floating to allow mismatch compliance of other sample preparation devices (Fig. 1 , 100) disposed within the cartridge (Fig. 3, 318).

[0073] Fig. 7B depicts the relative position of the actuator protrusion (522) and driving shaft (732) following mixing and piercing of the seal (Fig. 1 , 108) and prior to movement of the plunger (110) to expel the contents of the mixing chamber (Fig. 1 , 102). That is, after mixing and piercing, the driving shaft (732) may be rotated in a second direction as indicated by the arrow (738), which second direction is opposite the first direction. Responsive to the driving shaft (732) being rotated in the second direction, the protrusion (522) aligns with the slot in the driving shaft (732) such that the actuator (Fig. 1 , 106) disengages from the driving shaft (732) as depicted in Fig. 7B.

[0074] Then as depicted in Fig. 7C, the driving shaft (732) may press against the plunger (110) in a direction indicated by the arrow (736) such that the plunger (110) translates to push the sample out of the mixing chamber (Fig. 1 , 102) and into the fluid isolation chamber (Fig. 2, 214). Note that in this example, as the protrusion (522) of the actuator (Fig. 1 , 106) aligns with the slot, the actuator (Fig. 1 , 106) and mixer (104) remain translationally stationary while the plunger (110) moves in the direction indicated by the arrow (736).

[0075] Figs. 8A - 8C depict the translation of the plunger (110) to direct contents of the mixing chamber (102) to a downstream component, according to an example of the principles described herein. That is, as described above, following mixing, the mixer (104) and plunger (110) may be translated to 1) pierce, or otherwise open, the seal (108) and 2) drive the contents of the mixing chamber (102) to the fluid isolation chamber (Fig. 2, 214). [0076] Specifically, Fig. 8A depicts the actuator (106), which along with the driving shaft (Fig. 7, 732) allows spinning of the mixer (104) as well as translation of the mixer (104) and/or plunger (110). Fig. 8A also depicts the mixer (104) which is used to mix magnetic microparticles or a reagent with the sample. The mixing also aides in heating and cooling of the sample during lysing. As described above, in some examples, the mixer (104) may be used to open up the seal (108). That is, the seal (108) may be opened with the mixer (104) to direct the lysate from the mixing chamber (102) into the fluid isolation chamber (Fig. 2, 214).

[0077] Fig. 8A also depicts the interaction of the deflecting members (Fig. 5, 524) and the latches (Fig. 6, 628) to retain the mixer (104) to the actuator (106). In this example, the actuator (106) may spin freely within the plunger (110). [0078] As depicted in Fig. 8A, the mixing chamber (102) may house a reagent. In one particular example, the mixing chamber (102) includes lyophilized pellets (837) which may be stored in the mixing chamber (102) during manufacture.

[0079] The magnetic microparticles within the pellet (837) may be paramagnetic microparticles, superparamagnetic microparticles, diamagnetic microparticles, or a combination thereof, for example. As used in the present specification, the term magnetic microparticles may include microparticles that may not be magnetic in nature unless and until a magnetic field is introduced at a strength and proximity to cause them to become magnetic. The magnetic strength of the magnetic microparticles may be dependent on the magnetic field applied and may become stronger as the magnetic field is increased, or as the magnetic microparticles move closer to the magnetic source that is applying the magnetic field.

[0080] Specifically, paramagnetic microparticles may be those that have the ability to increase in magnetism when a magnetic field is present; however, paramagnetic microparticles are not magnetic when a magnetic field is not present. In some examples, the paramagnetic microparticles may exhibit no residual magnetism once the magnetic field is removed. A strength of magnetism of the paramagnetic microparticles may depend on the strength of the magnetic field, the distance between a source of the magnetic field and the paramagnetic microparticles, and a size of the paramagnetic microparticles.

[0081] Superparamagnetic microparticles may be similar to paramagnetic microparticles, however, they may exhibit magnetic susceptibility to a greater extent than paramagnetic microparticles in that the time it takes for them to become magnetized appears to be shorter. Diamagnetic microparticles may display magnetism due to a change in the orbital motion of electrons in the presence of a magnetic field.

[0082] As described above, the magnetic microparticles may be surface- activated to selectively bind with a biological component or may be bound to a biological component from a biological sample. In a specific example, an exterior of the magnetic microparticles may be surface-activated with interactive surface groups that can interact with a biological component of a biological sample or may include a covalently attached ligand.

[0083] In some examples, the ligand may include proteins, antibodies, antigens, nucleic acid primers, nucleic acid probes, amino groups, carboxyl groups, epoxy groups, tosyl groups, sulphydryl groups, or the like. In one example, the ligand may be a nucleic acid probe. The ligand may be selected to correspond with and to bind with the biological component. The ligand may vary based on the type of biological component targeted for isolation from the biological sample. For example, the ligand may include a nucleic acid probe when isolating a biological component that includes a nucleic acid sequence. In another example, the ligand may include an antibody when isolating a biological component that includes antigen.

[0084] In some examples, the magnetic microparticles can have an average particle size ranging from 10 nanometers (nm) to 50,000 nm. In yet other examples, the magnetic microparticles may have an average particle size ranging from 500 nm to 25,000 nm, from 10 nm to 1 ,000 nm, from 25,000 nm to 50,000 nm, or from 10 nm to 5,000 nm. As used in the present specification, the term “average particle size" describes a diameter or average diameter, which may vary, depending upon the morphology of the individual particle. A shape of the magnetic microparticles may be spherical, irregular spherical, rounded, semi-rounded, discoidal, angular, sub-angular, cubic, cylindrical, or any combination thereof. The shape of the magnetic microparticles may be spherical and uniform, which may be defined herein as spherical or near- spherical, e.g., having a sphericity of >0.84. Thus, any individual particles having a sphericity of <0.84 are considered non-spherical (irregularly shaped). [0085] As depicted in Fig. 8A, the mixing chamber (102) may include a neck- down region near the mixer (104) head. Such a region increases the fluid height for heating. That is, were the mixing chamber (102) wider at this point, the fluid level would be lower and the volume of fluid to be heated (i.e., the space between the mixier (104) head and the walls of the mixing chamber (102)) would be greater, thus resulting in a less efficient heating of the sample. [0086] Fig. 8A also depicts the plunger (110) with the O-ring seals (420-1 , 420-2). The O-ring seals (420) may be made of a deformable and elastic material that create a seal between the plunger (110) and the mixing chamber (102) walls, but allows for translation of the plunger (110). That is, the O-ring seals (420-1 , 420-2) seal the mixing chamber (102) during mixing and plunging. The plunger (110) pushes air and lysate out during dispense. The O-ring seals (420-1 , 420-2) by creating a seal prevent air/fluid escape such that a pressure may be used to expel the contents of the mixing chamber (102) towards the fluid isolation chamber (Fig. 2, 214).

[0087] In an example, the combined length of the plunger (110) and the actuator (106) is to fit within the mixing chamber (102) but not block the input port (212) opening during lysing. The plunger (110) and actuator (106) may block the passage from the input port (212) to the mixing chamber (102) following plunging as depicted in Figs. 8B and 8C.

[0088] Within the mixing chamber (102), a volume of air may be provided above the sample. Air below the input port (212) is to exit into the fluid isolation chamber (214). In some examples, the volume of air provided in the mixing chamber (102) is the same as the amount of sample lysate. In another example, it may be 3.5 times the amount of sample lysate. The air may act as a buffer and prevent fluid contact with the plunger (110) and O-ring seals (420). Doing so prevents fluid from being retained in the mixing chamber (102). That is, any sample left in the mixing chamber (102) is not available for downstream analysis. Accordingly, the air buffer prevents any sample from becoming stranded on the plunger (110) rather than being delivered to the fluid isolation chamber (214).

[0089] In an example, the mixer (104) head is close to the film, for example, between 0.1 and 1.5 millimeters above the seal (108). Doing so increases the efficiency of mixing. That is, the magnetic microparticles may settle. Any magnetic microparticles that settle below the mixer (104) may not be under the influence of the mixer (104) and thus may not mix with the sample. Accordingly, placing the mixer (104) close to the seal (108) reduces the space wherein magnetic microparticles may settle or otherwise be inactive. As described above, Fig. 8A depicts the state of the sample preparation device (100) following mixing, but prior to piercing.

[0090] During piercing, as depicted in Fig. 8B, the protrusion (Fig. 5, 522) is aligned with the notch in the driving shaft (Fig. 7, 732) as depicted in Fig. 7A, and the driving shaft (Fig. 7, 732) is driven down as indicated by the arrow (736). Doing so pushes the mixer (104) to pierce the seal (108) providing a fluidic path between the mixing chamber (104) and the fluid isolation chamber (Fig. 2, 214). Note that as depicted in Fig. 8B, the input port (212) remains partially open during piercing.

[0091] Once pierced, the driving shaft (Fig. 7, 732) is rotated as depicted in Fig. 7B to disengage the actuator (106) from the driving shaft (Fig. 7, 732). The driving shaft (Fig. 7, 732) may then be driven in the direction indicated by the arrow (736). As the actuator (106) and mixer (104) are no longer engaged with the driving shaft (Fig. 7, 732), they remain stationary. However, the driving shaft (Fig. 7, 732) may still press against the plunger (110) moving it downwards to evacuate the mixing chamber (102) as depicted in Fig. 8C. That is, when the protrusion (Fig. 5, 522) aligns with the slot and responsive to a translational force, the plunger (110) translates to dispel contents of the mixing chamber (102).

[0092] During fluid ejection, the input port (212) is sealed by the plunger (110) such that the air cushion pushes the lysate out the opened seal (108). Also, in this example, the second O-ring seal (420-2) seals the mixing chamber (102) preventing air/fluid escaping through the input port (212) and preventing air/fluid from entering the mixing chamber (102) through the input port (212). That is, in the evacuated state where the sample has been drained from the mixing chamber (102), the plunger (100) blocks the input port (212) of the mixing chamber (102). Note that in this example, the plunger (110) spans the input port (212). In an example, the second O-ring seal (420-2) is larger than the input port (212) opening so as to not be pinched within the input port (212) during translation of the plunger (110).

[0093] Fig. 9 is a cross-sectional view of the sample preparation device (100), according to another example of the principles described herein. Fig. 9 depicts an example, where the plunger (Fig. 1 , 110), actuator (Fig. 1 , 106), and mixer (Fig. 1 , 104) are a single integrated component. That is, rather than having an actuator (Fig. 1 , 106) that is affixed with the mixer (Fig. 1 , 104) that slides through the plunger (Fig. 1 , 110), these three components may be integrated. Fig. 9 also depicts an example, where the seal (Fig. 1 , 108), rather than being a film seal, is a ball (940) or cork. In this example, the action of the mixer (Fig. 1 , 104) or air pressure resulting from the downward motion of the mixer (Fig. 1 , 104), pushes the ball (940) out of a throat. As depicted in Fig. 9, the ball (940) may be free to drop into the fluid isolation chamber (Fig. 2, 214) or as depicted in Fig. 10 may be retained in a catch (1042) so as to not block fluid flow through the remaining portions of the sample preparation device (100).

[0094] Fig. 10 is a cross-sectional view of the sample preparation device (100), according to another example of the principles described herein. In this example, the seal (Fig. 1 , 108) is a ball (940) that upon displacement is retained in a catch (1042).

[0095] Fig. 11 is a cross-sectional view of the seal (Fig. 1 , 108) of the sample preparation device (100), according to an example of the principles described herein. In the example depicted in Fig. 11 , the seal (Fig. 1 , 108), rather than being a film seal, is a plug (1142). In this example, the action of the mixer (Fig. 1 , 104) or air pressure resulting from the downward motion of the mixer (Fig. 1 , 104), dislodges the plug (1142) out of a throat. As with the ball (Fig. 9, 942), the plug (1142) may be tethered to the housing of the sample preparation device (100) or may be free to fall into the fluid isolation chamber (Fig. 2, 214) following dislodgement.

[0096] Fig. 12 is a cross-sectional view of the seal (Fig. 1 , 108) of the sample preparation device, according to an example of the principles described herein. In the example depicted in Fig. 12, the seal (Fig. 1 , 108), rather than being a film seal, is a septum (1244), which may or may not have a slit. In this example, the action of the mixer (Fig. 1 , 104) or air pressure resulting from the downward motion of the mixer (Fig. 1 , 104) opens the septum (1244). Fig. 12 also depicts the pellet (837) that may include the magnetic microparticles.

[0097] Fig. 13 is a cross-sectional view of the seal (Fig. 1 , 108) of the sample preparation device, according to an example of the principles described herein. In the example depicted in Fig. 13, the seal (Fig. 1 , 108), rather than being a film seal, is a pipette tip (1346) to be acted upon by the mixer (104) to allow contents of the mixing chamber (102) to pass. That is, the pipette tip (1346), when pushed down by the mixer (104), may open/unseal a septum, film, or other seal. The channel through the pipette tip (1346) provides a fluid flow and thereby increase fluid flow from the mixing chamber (102) to the fluid isolation chamber (Fig. 2, 214). Fig. 13 also depicts the pellet (837) that may include the magnetic microparticles.

[0098] Fig. 14 is a cross-sectional view of the seal (Fig. 1 , 108) of the sample preparation device (100), according to an example of the principles described herein. In this example, the seal (Fig. 1 , 108) is ruptured by a needle (1448) affixed to a bottom of the mixing chamber (Fig. 1 , 102). In this example, the mixer (104) may be translated down. The downward pressure may cause a seal (108) to translate downwards towards the needle (1448). Accordingly, when pushed sufficiently down, the needle (1448) ruptures the seal (108). The needle (1448) may include a hole such that when protruding through the seal (108), fluid within the mixing chamber (102) passes through the hole in the needle (1448) into the fluid isolation chamber (Fig. 2, 214). Fig. 14 also depicts the pellet (837) that may include the magnetic microparticles. [0099] Fig. 15 is a cross-sectional view of the seal (Fig. 1 , 108) of the sample preparation device, according to an example of the principles described herein. In the example depicted in Fig. 15, the seal includes multiple sliding seals (108- 1 , 108-2, 108-3) that are translated to expose a bypass channel (1550) to allow contents of the mixing chamber (102) to pass. For example, as the mixer/plunger translate to move the seal, the seals (108) translate down, to provide a fluid path, i.e. , a bypass channel (1550), that allows the fluid to flow from the mixing chamber (102) towards the fluid isolation chamber (104). [00100] Figs. 16A and 16B are cross-sectional views of the sample preparation device (100) according to another example of the principles described herein. In the example depicted in Figs. 16A and 16B, the plunger (Fig. 1 , 110) and actuator (Fig. 1 , 106) are a single integrated component. The plunger/actuator integrated component provides the rotational motion to the mixer (104). Accordingly, the mixer (104) may be keyed to match a cross- sectional area of the plunger shaft as depicted in Fig. 16B. Accordingly, as the plunger rotates, the keyed interface between the plunger and mixer (Fig. 1 , 104) cause the mixer (Fig. 1 , 104) to rotate and aide in lysing and binding.

[00101] In this example, the plunger shaft slides through the mixer (104) opening to open the seal (Fig. 1 , 108). That is, the plunger/actuator integrated component slides through the sleeve in the mixer (104) depicted in Fig. 16B to open the seal (Fig. 1 , 108). In other words, the shaft protrudes out the bottom of the mixer (104) to open the seal. Note that while Figs. 16A and 16B depict a ball (940) seal, other seals (Fig. 1 , 108) such as those described herein may be implemented in accordance with the principles described herein.

[00102] Fig. 17 is a flow chart of a method (1700) of sample preparation, according to an example of the principles described herein. As described above, sample preparation may include lysing a sample and binding contents of a lysate, i.e., nucleic acid, to magnetic microparticles or facilitating an interaction between the sample and a reagent.

[00103] The reagent that is mixed with the sample may take many forms. For example, the reagent may include water, some salts to control pH (such as Tris hydrochloride), other salts (such as magnesium chloride (MgCI2), lithium chloride (LiCI), and sodium chloride (NaCI)) to aid in the binding reaction, a surfactant to help with heat lysing and to keeping magnetic microparticles distributed and to aid in sample input liquid flow, and some preservatives or biocides for reagent storage.

[00104] In some examples, the reagent may include alcohols (ethanol or isopropanol) that help with lysing and binding; chaotropic salts (such as guanidine hydrochloride or guanidine thiocyanate) that help with denaturing proteins from the sample, lysing microbes, binding nucleic acids, more aggressive surfactants for lysing potentially without as much heat (sodium dodecyl sulfate (SDS), LiDS, sodium lauroyl sarcosinate, etc.), and other denaturants such as Proteinase K, dithiothreitol (DTT), etc.

[00105] In this example, the sample is lysed (block 1701 ) by rotating a mixer (Fig. 1 , 104) of a sample preparation device (Fig. 1 , 100) disposed within the host station. That is, the sample preparation device (Fig. 1 , 100) along with multiple other sample preparation devices (Fig. 1 , 100) may be placed in a cartridge (Fig. 3, 318) which is inserted into a host station. The host station may include multiple driving shafts (Fig. 7, 732) each to interact with a different sample preparation device (Fig. 1 , 100). The driving shafts (Fig. 7, 732) rotate and via the actuator (Fig. 1 , 106)/driving shaft (Fig. 7, 732) interface, and a actuator (Fig. 1 , 106)/mixer (Fig. 1 , 104) interface, transmit (block 1701) the rotational force to the mixer (Fig. 1 , 104) which mixer (Fig. 1 , 104) agitates the sample to ensure thorough mixing, and/or lysing and binding.

[00106] Following mixing, a seal (Fig. 1 , 108) between the mixing chamber (Fig. 1 , 102) and the fluid isolation chamber (Fig. 2, 214) is ruptured (block 1702). This may be done by translating the driving shaft (Fig. 7, 732) when the actuator protrusions (Fig. 5, 522) are positioned within notches of the driving shaft (Fig. 7, 732) as depicted in Fig. 7 A.

[00107] The contents of the mixing chamber (Fig. 1 , 102) are then driven (block 1703) towards the fluid isolation chamber (Fig. 2, 214). Specifically, the plunger (Fig. 1 , 110) may be translated (block 1703) through the mixing chamber (Fig. 1 , 102) to drive contents of the mixing chamber (Fig. 1 , 102) towards the fluid isolation chamber (Fig. 2, 214). That is, the driving shaft (Fig. 7, 732) may be rotated in an opposite direction to disengage the protrusions (Fig. 5, 522) from the notch and instead align the protrusions (Fig. 5, 522) with a slot in the driving shaft (Fig. 7, 732). A translational force upon the plunger (Fig. 1 , 110) drives the plunger (Fig. 1 , 110) to expel the contents of the mixing chamber (Fig. 1 , 102) to the fluid isolation chamber (Fig. 2, 214) while the actuator (Fig. 1 , 106)/mixer (Fig. 1 , 104) remain stationary.

[00108] Fig. 18 depicts an interaction of a driving shaft (732) with the sample preparation device (Fig. 1 , 100), according to an example of the principles described herein. As described above, the host station may include a driving shaft (732) that is coupled to a motor which provides rotational movement. In this example, the driving shaft (732) includes a T-shaped slot. During use, the sample preparation device (Fig. 1 , 100) may be aligned such that the protrusion (522) on a actuator (Fig. 1 , 106) or a mixer (Fig. 1 , 104) shaft aligns with the slot. When the sample preparation device (Fig. 1 , 100) is fully seated, that is when an end of the plunger (Fig. 1 , 110) contacts an end surface of the driving shaft (732), the driving shaft (732) may be rotated as indicated by the arrow (1842). This causes the protrusion (522) to align with, or sit in, a crossbar of the T-slot. Due to this interface between the protrusion (522) and the crossbar, as the driving shaft (732) is rotated in the direction indicated by the arrow (1842), the actuator (Fig. 1 , 106) and/or mixer (Fig. 1 , 104) are also rotated in the direction indicated by the arrow (1842).

[00109] The interface between the T-shaped slot and the protrusion (522) may also transmit the translational force. That is, when the protrusion (522) is seated in the crossbar and responsive to a translational force from the driving shaft (732) against the actuator (Fig. 1 , 106) or mixer (Fig. 1 , 104), the actuator (Fig. 1 , 106) and/or mixer (Fig. 1 , 104) move to open the seal.

[00110] Such a T-shaped slot allows for a pushing and pulling motion while the mixer and/or actuator is hooked into the T. Such may be used for example, with a linear mixer and/or to retract the mixer (Fig. 1 , 104) and actuator (Fig. 1 , 106) following plunging.

[00111] In summary, using such a sample preparation device 1 ) enables automated lysing of cells and binding components such as nucleic acids to magnetic microparticles; 2) maintains a liquid sealed chamber during mixing/heating for minimal contamination risk during sample preparation where air can escape but not liquid; 3) allows for controlled delivery of the lysate to the fluid isolation chamber; 4) enables multiple instances of the sample preparation to occur in the same physical space; 5) executes one sample transfer operation;

6) reduces stranded fluid; and 7) remains sealed for disposal. However, the devices disclosed herein may address other matters and deficiencies in a number of technical areas.

Claims

CLAIMS What is claimed is:

1 . A sample preparation device, comprising: a mixing chamber to receive a sample; a mixer disposed within the mixing chamber to mix the sample; an actuator to transmit a torque to the mixer; a seal to separate the mixing chamber from a downstream component; and a plunger disposed within the mixing chamber to direct contents of the mixing chamber to the downstream component.

2. The sample preparation device of claim 1 , wherein a mixer shaft aligns longitudinally with the mixing chamber and passes through the plunger.

3. The sample preparation device of claim 1 , wherein: the mixer is attached to the actuator; the actuator comprises a protrusion to interface with a notch in a driving shaft of a host station such that as the driving shaft rotates in a first direction, the actuator and mixer rotate in the first direction; and when the protrusion aligns with the notch, and responsive to a translational force against the actuator, the actuator is to move the mixer to open the seal.

4. The sample preparation device of claim 3, wherein responsive to the driving shaft being rotated in a second direction that is opposite the first direction, the protrusion aligns with a slot in the driving shaft such that the actuator disengages from the driving shaft.

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5. The sample preparation device of claim 4, wherein when the protrusion aligns with the slot, and responsive to a translational force, the plunger translates to dispel contents of the mixing chamber.

6. The sample preparation device of claim 1 , wherein: the mixer is attached to the actuator; the actuator comprises a protrusion to interface with a T-shaped slot in a driving shaft of a host station such that as the driving shaft rotates in a first direction, the actuator and mixer rotate in the first direction; and when the protrusion aligns with the crossbar of the T-shaped slot, and responsive to a translational force against the actuator, the actuator is to move the mixer to open the seal.

7. The sample preparation device of claim 1 , wherein when in an evacuated state, the plunger is to block an input port to the mixing chamber.

8. The sample preparation device of claim 1 , wherein the seal is selected from the group consisting of: a film; a ball; a plug; a septum; a pipette tip to be acted upon by the mixer to allow contents of the mixing chamber to pass; a sliding seal to be translated to expose a bypass channel to allow contents of the mixing chamber to pass; and a seal to be ruptured by a needle affixed to a bottom of the mixing chamber.

9. A method, comprising: lysing a sample in a mixing chamber by rotating a mixer of a sample preparation device disposed within a host station; rupturing a seal between a mixing chamber of the sample preparation device and a fluid isolation chamber of the sample preparation device; and driving contents of the mixing chamber towards the fluid isolation chamber by translating a plunger through the mixing chamber.

10. The method of claim 9, wherein the seal is ruptured via a mechanical interaction or via air pressure.

11 . The method of claim 9, further comprising at least one of: binding contents of a lysate to magnetic microparticles; and facilitating an interaction between the sample and a reagent.

12. A sample preparation device, comprising: a mixing chamber to receive a sample; an input port to introduce the sample into the mixing chamber; a mixer disposed within the mixing chamber to mix the sample; a actuator to transmit a torque to the mixer; a film seal to separate the mixing chamber from a downstream component; a plunger disposed within the mixing chamber to direct contents of the mixing chamber to the downstream component; a fluid isolation chamber to receive the contents and to house a reaction; and an output to dispense contents of the fluid isolation chamber.

13. The sample preparation device of claim 12, wherein the sample preparation device is disposed in a cartridge along with other sample preparation devices.

14. The sample preparation device of claim 12, wherein the plunger, actuator, and mixer are a single integrated component.

15. The sample preparation device of claim 12, wherein: the plunger and actuator are a single integrated component; and a plunger shaft is to slide through a mixer opening to rupture the film seal.