High throughput biological assay systems and methods

Abstract

A biological assay system includes a vessel for receiving a biological sample, a temperature control unit to regulate the temperature within the vessel, and a carrier designed to move the vessel close to the temperature control unit.

Description

Docket No. G10083312P1PCTHIGH THROUGHPUT BIOLOGICAL ASSAY SYSTEMS AND METHODSCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority under 35 U.S.C. § 119(e) and 37 C.F.R. § 1.78 to provisional application no. 63 / 708,099 filed on October 16, 2024, titled “HIGH THROUGHPUT BIOLOGICAL ASSAY SYSTEMS AND METHODS " which is hereby incorporated by reference herein in its entirety.BACKGROUND

[0002] Polymerase Chain Reaction (PCR) is a technique for amplifying nucleic acid sequences (e.g., DNA, RNA, mRNA, etc...). Traditional methods have several challenges that limit their effectiveness and applicability. Traditional PCR methods require precise addition of reagents at certain times and repeated cycling (e.g., 20-50 times) of a sample between different temperature ranges. The samples are typically cyclically heated and cooled tens of times to achieve the proper reaction, thereby resulting in slow process times, increased costs, and limited throughput.BRIEF SUMMARY

[0003] A biological assay system includes: a vessel configured to receive a biological sample; a temperature control unit; and a carrier configured to move the vessel proximate to the temperature control unit to control a temperature of the biological sample within the vessel.

[0004] Optionally, in some embodiments, the vessel includes a thin-walled cylindrical element.

[0005] Optionally, in some embodiments, a wall thickness of the vessel ranges from 0.10 mm to 0.35 mm.

[0006] Optionally, in some embodiments, the biological sample has a volume of less than 2 pL.

[0007] Optionally, in some embodiments, the vessel has a surface area of contact with the temperature control unit of between 5 and 20 mm2.

[0008] Optionally, in some embodiments, the vessel includes a hypodermic needle.

[0009] Optionally, in some embodiments, the actuator includes a stepper motor.Docket No. G10083312P1PCT

[0010] Optionally, in some embodiments, the system further includes a second temperature control unit and a third temperature control unit. The actuator moves the vessel between the temperature control units to perform a polymerase chain reaction (PCR) process.

[0011] Optionally, in some embodiments, the temperature control unit is configured to achieve a temperature change rate in the sample up to 2000 °C per second.

[0012] Optionally, in some embodiments, the carrier is configured to couple to a plurality of vessels and move each of the plurality of vessels proximate to the temperature control unit.

[0013] Optionally, in some embodiments, the system further includes a light sensor configured to detect fluorescence generated by the biological sample in response to completion of a chemical reaction.

[0014] A biological assay method includes providing a vessel containing a biological sample; providing a temperature control unit; moving the vessel proximate to the temperature control unit to control a temperature of the biological sample within the vessel.

[0015] Optionally, in some embodiments, the method further includes performing an operation of a PCR process in the vessel at the temperature.

[0016] Optionally, in some embodiments, the method further includes a denaturation operation and the temperature control unit maintains the temperature in the range of 94 °C to 98 °C.

[0017] Optionally, in some embodiments, the method further includes an annealing operation and the temperature control unit maintains the temperature in the range 50 °C to 65 °C.

[0018] Optionally, in some embodiments, the method further includes an extension operation and the temperature control unit maintains the temperature in the range 72 °C to 75 °C.

[0019] Optionally, in some embodiments, the biological sample has a volume of less than 0.5 pL.

[0020] Optionally, in some embodiments, the vessel has a surface area of contact with the temperature control unit of between 5 and 20 mm2.

[0021] Optionally, in some embodiments, the method further includes moving the vessel proximate to a second temperature control unit and a third temperature control unit, wherein the actuator moves the vessel between the temperature control units to perform a polymerase chain reaction (PCR) process.Docket No. G10083312P1PCT

[0022] A biological assay system includes a plurality of needles configured to receive a respective plurality of biological samples; a plurality of temperature control units; and a carrier configured to receive the plurality of needles and to move the plurality of needles proximate to the plurality of temperature control units, such that each temperature control unit of the plurality of temperature control units controls a temperature of the plurality of biological samples.

[0023] In one embodiment, a high-throughput biological assay system includes: a printed circuit board; plurality of capacitive pads disposed on the printed circuit board and configured to generate electrostatic forces to move an aqueous sample containing a biological sample across a surface of the printed circuit board; a plurality of heaters embedded within the printed circuit board, each heater corresponding to a temperature zone; and a dielectric layer positioned over the capacitive pads and configured to separate the capacitive pads from the aqueous sample while enabling the electrostatic forces to act on the aqueous sample.

[0024] Optionally in some embodiments, the system further includes a plurality of temperature sensors positioned adjacent to the respective heaters to monitor temperature conditions within each temperature zone.

[0025] Optionally in some embodiments, the plurality of capacitive pads comprises a first capacitive pad, a second capacitive pad, a third capacitive pad, and a fourth capacitive pad arranged to enable sequential movement of the aqueous sample between temperature zones.

[0026] Optionally in some embodiments, the system further includes a sample source configured to supply the aqueous sample for electrostatic transfer to the capacitive pads.

[0027] Optionally in some embodiments, the plurality of heaters comprises a first heater configured to maintain a temperature suitable for denaturation, a second heater configured to maintain a temperature suitable for annealing, and a third heater configured to maintain a temperature suitable for extension.

[0028] Optionally in some embodiments, the system further includes at least one aperture extending through the printed circuit board to allow optical access to the aqueous sample.

[0029] Optionally in some embodiments, the system further includes a fluorescence sensor positioned adjacent to the aperture to detect fluorescence signals generated by the biological sample.

[0030] Optionally in some embodiments, the dielectric layer has a thickness of approximately 10 micrometers and is formed from a transparent fluorinated material.

[0031] Optionally in some embodiments, the system further includes a cap layer positioned above the dielectric layer to prevent evaporation of the aqueous sample during processing.Docket No. G10083312P1PCT

[0032] Optionally in some embodiments, the capacitive pads are configured to apply alternating voltages that utilize polar characteristics of water molecules within the aqueous sample to generate attractive forces for sample movement.

[0033] Optionally in some embodiments, system is configured to perform a polymerase chain reaction process by moving the aqueous sample sequentially through a first zone for denaturation, a second zone for annealing, and a third zone for extension.BRIEF DESCRIPTION OF THE DRAWINGS

[0034] FIG. 1 A is a schematic of a high-throughput biological assay system in a first configuration.

[0035] FIG. IB is a schematic of the high-throughput biological assay system of FIG. 1A in a second configuration.

[0036] FIG. 2A is a schematic of an embodiment of the high-throughput biological assay system of FIG. 1A in a first configuration.

[0037] FIG. 2B is a schematic of an embodiment of the high-throughput biological assay system of FIG. 1A in a second configuration.

[0038] FIG. 2C is a schematic of an embodiment of the high-throughput biological assay system of FIG. 1A in a third configuration.

[0039] FIG. 3A is a perspective view of an embodiment of the high-throughput biological assay system of FIG. 1A.

[0040] FIG. 3B is a partially exploded perspective view of the high-throughput biological assay system of FIG. 3 A.

[0041] FIG. 3C is sectioned, perspective view of a carriage of the high-throughput biological assay system of FIG. 3A taken along line 3C-3C of FIG. 3B.

[0042] FIG. 3D is sectioned, elevation view of the carriage of the high-throughput biological assay system of FIG. 3 A taken along line 3C-3C of FIG. 3B.

[0043] FIG. 3E is sectioned, perspective view of a temperature control unit of the high- throughput biological assay system of FIG. 3 A taken along line 3D-3D of FIG. 3B.

[0044] FIG. 3F is sectioned, elevation view of a temperature control unit of the high- throughput biological assay system of FIG. 3 A taken along line 3D-3D of FIG. 3B.

[0045] FIG. 3G is a front elevation view of the high-throughput biological assay system of FIG. 3A in a first configuration.Docket No. G10083312P1PCT

[0046] FIG. 3H is a front elevation view of the high-throughput biological assay system of FIG. 3A in a second configuration.

[0047] FIG. 4A is a plan view of an embodiment of a heater of the high-throughput biological assay system of FIG. 3A.

[0048] FIG. 4B is a plan view of an embodiment of a sensor array of the high-throughput biological assay system of FIG. 3 A.

[0049] FIG. 5 is a flow chart of a method of operating the high-throughput biological assay system of FIG. 1A. S

[0050] FIG. 6 is a plan view of an embodiment of the high-throughput biological assay system with capacitive pads and heaters.

[0051] FIG. 7A is an elevation view schematic of the high-throughput biological assay system of FIG. 6 with a sample being drawn from a sample source.

[0052] FIG. 7B is an elevation view schematic of the high-throughput biological assay system of FIG. 6 with the sample moved to a first zone for denaturation.

[0053] FIG. 7C is an elevation view schematic of the high-throughput biological assay system of FIG. 6 with the sample moved to a second zone for annealing.

[0054] FIG. 7D is an elevation view schematic of the high-throughput biological assay system of FIG. 6 with the sample moved to a third zone for extension.

[0055] FIG. 8 is a simplified block diagram of components of a computing system of the high-throughput biological assay system of FIG. 3A.DETAILED DESCRIPTION

[0056] The high-throughput biological assay systems and methods disclosed enable rapid processing of multiple biological samples with precise temperature control. The systems move one or more sample materials through one or more temperature zones adapted to support various reactions. The systems enable high temperature rates of change in liquid samples, thereby supporting rapid, parallel processing of multiple biological samples.

[0057] The disclosed systems include an actuator that moves a vessel containing a biological sample through one or more temperature zones to promote desired reactions. In some examples, the sample is carried in a vessel such as a standard hypodermic or blunt, hollow needle. The small volume of liquid, thin wall of a needle, and relatively high heat transfer area of the needle relative to the mass of sample enable high rates of temperature change of the sample, and thus high processing speed relative to standard methods.Docket No. G10083312P1PCT

[0058] Turning to the figures, FIG. 1A and FIG. IB show schematics of an embodiment of a high-throughput biological assay system 100. The high-throughput biological assay system 100 includes an actuator 102, a vessel 104, and at least one process zone 106. In the example shown, the high-throughput biological assay system 100 includes an optional second zone 108. More than two zones may be used as desired. In some embodiments, the high-throughput biological assay system 100 includes multiple actuators 102 and / or multiple vessels 104. Where multiple vessels 104 are used, the vessels 104 may be moved by one actuator 102 or by multiple actuators 102.

[0059] The vessel 104 is adapted to receive a sample 110, typically a liquid sample, but the sample 110 may also be a solid, gas, or combinations of solids, liquids, and / or gases. The vessel may be any container suitable to contain the sample, in whatever form of matter the sample is, and enable a stimulus from the zone to reach and affect the sample 110. For example, where the stimulus is heat energy, the vessel is configured to enable heat to pass from the zone 106 into the sample 110. In examples where the stimulus is light, the vessel enables the transmission of light from the zone 106 to the sample 110, and so on.

[0060] The actuator 102 moves the vessel 104, and thus the sample 110 into or out of the zone 106, and if present, the optional zone 108. Any zone of the present disclosure may subject the sample 110 to a stimulus such as heat, cold, radiation, dark, light, vibration, etc. In many embodiments, the stimulus is the application of heat to the vessel 104, the zone is a temperature control unit that transmits heat to the sample 110 to effectuate a chemical or biological reaction, such as a polymerase chain reaction, or other reaction.

[0061] As shown for example in FIG. 1A and FIG. IB, the actuator 102 moves the vessel 104 and the sample 110 between the zone 106 and the zone 108. In embodiments where the zone 108 is not included, the actuator 102 moves the sample 110 in and out of the zone 106, subjecting the sample 110 to the ambient environment as a stimulus when the vessel 104 is not in the zone 106.

[0062] FIG. 2A through FIG. 2C illustrate schematics of an embodiment of high- throughput biological assay system 200. The high-throughput biological assay system 200 is similar to the high-throughput biological assay system 100 in many respects. For example, the high-throughput biological assay system 200 includes at least two zones, and may include a third or more zones (e.g., a first zone 202, a second zone 204, or a third zone 206). Like the high-throughput biological assay system 100, the high-throughput biological assay system 200 also includes a vessel, in this case a needle or other relatively thin container. The vessel 208 is configured to hold a sample 210, which may be the same as aDocket No. G10083312P1PCT sample 110. The high-throughput biological assay system 200 also includes an actuator 102. configured to introduce movement to the vessel.

[0063] The high-throughput biological assay system 200 is a specific embodiment of the high-throughput biological assay system 100 where in this embodiment the vessel 208 is a thin-walled (e.g., the wall thickness may range from approximately 0.10 mm to 0.35 mm) cylindrical element like a needle. In some embodiments, the sample 210 volume may be in the inclusive range of about 0.5 to 50 pL. In some embodiments, a sample 210 may occupy only a portion of the inner volume of the needle and thus may be smaller than this range (e.g., the volume may be less than 0.5 pL). The small sample 210 volume, thin wall, and high surface area (relative to the sample 210 volume) enable the rapid change of temperature of the samples 210. For example, temperature change rates of up to 2000 °C per second or more are achievable with the high-throughput biological assay system 200. The sample 210 may be as small as a drop of liquid disposed in the vessel 208 or may be a larger volume as needed for the desired testing. The sample 210 may be held in a lumen 212 of the vessel 208 by surface tension or a vacuum. In some embodiments, the vessel 208 may be sealed, whereas in other embodiments, the vessel 208 may be open at one or both ends of the lumen 212.

[0064] The high-throughput biological assay system 200 may be adapted to perform a polymerase chain reaction (PCR) process. PCR typically involves three process steps with reagents processed at different temperatures. For example, a PCR process may include one or more of the following steps:Denaturation

[0065] In denaturation, double-stranded deoxyribonucleic acid (DNA) is separated into two single strands, allowing each to serve as a template for synthesis in subsequent steps. During denaturation, the double-stranded DNA in the biological sample is heated to break the hydrogen bonds holding the two strands together. This heating results in the separation of the DNA strands, creating single-stranded DNA templates. The denaturation step typically occurs at a temperature range of 94 °C to 98 °C. The precise temperature may vary depending on the specific PCR protocol and the composition of the DNA being amplified.Annealing

[0066] In the annealing step of PCR, primers (short single-stranded sequences of nucleotides designed to match regions flanking the target DNA sequence) bind, or anneal, to their complementary sequences on the single-stranded DNA templates created duringDocket No. G10083312P1PCT denaturation. This step enables the primers to attach to the appropriate locations on the template DNA. The temperature of the reaction mixture is lowered relative to the denaturation step to allow the primers to form hydrogen bonds with their complementary sequences on the single-stranded DNA templates. The annealing step typically occurs at a temperature range between 50 °C and 65 °C. The temperature may depend on the melting temperature of the primers, which is influenced by their length, composition, and other properties.Extension (or Elongation)

[0067] In the extension (or elongation) step of PCR, DNA polymerase (an enzyme) synthesizes a new strand of DNA complementary to the DNA template strands generated by the annealing process by adding nucleotides to the primer. The DNA polymerase attaches to the primers bound to the single-stranded DNA templates. The polymerase extends the primers by adding nucleotides to the end of the primer, synthesizing the new strand of DNA complementary to the template strand. This results in the formation of new double-stranded DNA starting from the primers. The extension step typically occurs at a temperature range between 72 °C to 75 °C.

[0068] Each of these three steps, denaturation, annealing, and extension may be repeated tens to hundreds of times on a single sample to achieve sufficient amplification of the target DNA.

[0069] The high-throughput biological assay system 200 is adapted to move the sample 210 through the first zone 202, the second zone 204, and the third zone 206 and back, where each zone is configured to control the sample 210 temperature to a desired level.

[0070] For example, the first zone 202 may be adapted to bring a sample 210 to the temperature range suitable for the denaturation process (e.g., about 94 °C - 98 °C). Similarly, the second zone 204 may be adapted to bring the sample 210 to the temperature range suitable for annealing (e.g., between 50 °C and 65 °C). The third zone 206 may be adapted to bring the sample 210 to the temperature suitable for extension (e.g., between 72 °C to 75°C).

[0071] The actuator 102 may move the vessel 208 and thus the sample 210 proximate to or into the first zone 202 to perform the denaturation step, to the second zone 204 to perform the annealing step, and then to the third zone 206 to perform extension. The actuator may return the sample 210 to the first zone 202 and the process begins again. The process may end when a desired level of DNA amplification has been achieved. Because the walls of the vessel 208 are thin and the sample 210 is small, the sample 210 reaches the desiredDocket No. G10083312P1PCT temperature rapidly (e.g., changing at about 2000 °C per second). Thus, the high-throughput biological assay system 200 can cycle through the three steps of a PCR process, or other biological process extremely rapidly compared to prior methods.

[0072] With reference to FIG. 3A through FIG. 4B, a high-throughput biological assay system 300 is described. The high-throughput biological assay system 300 is a specific implementation of the high-throughput biological assay system 100 and the high-throughput biological assay system 200. The high-throughput biological assay system 300 includes a first temperature control unit 302 (omitted from FIG. 3A for the sake of clarity), a second temperature control unit 304, and a third temperature control unit 306, a drive assembly 308, a carrier 314, and a frame 320. Like the high-throughput biological assay system 100 and the high-throughput biological assay system 200, the high-throughput biological assay system 300 moves a sample 110, 210 in and out of one or more zones or temperature control units. The high-throughput biological assay system 300, like the high-throughput biological assay system 200 is adapted to perform a PCR process, as described with respect to the high-throughput biological assay system 200.

[0073] With specific reference to FIG. 3 A and FIG. 3B, the major components and subsystems of the high-throughput biological assay system 300 are shown. A single vessel 312 and single first temperature control unit 302 are shown, for simplicity. The vessel 312 is a standard hypodermic needle (preferably a blunt needle) with a needle shaft 330 and a flange 328. The vessel 312 is coupleable to the carrier 314. The carrier 314 is moveable with respect to the first temperature control unit 302 by the drive assemblies 308. The first temperature control unit 302 includes a circuit board with heater traces therein that heat the needle shaft 330, and this the sample, as the shaft is passed into the temperature control unit by the carrier 314.

[0074] The high-throughput biological assay system 300 includes a pair of drive assemblies 308, each including an actuator 318. See also FIG. 3G and FIG. 3H. The drive assemblies 308 include an actuator 318, supported on a spine 350. In this example, the actuator 318 is a stepper motor. In other embodiments, the actuator 102 may be another suitable device that can move the carrier 314 with respect to the temperature control units. For example, the actuator 318 may be a servo motor, dashpot, hydraulic or pneumatic cylinder / piston, roller chain and sprocket, belt, or the like.

[0075] The actuator 318 is coupled to a screw 332 via a coupler 338. The coupler 338 may be a flexible device that transmits torque from the actuator 318 to the screw 332 yet allows for slight misalignment of the screw 332 and actuator 318 or absorbs shocks therebetween.Docket No. G10083312P1PCT

[0076] The drive assembly 308 includes a guide 342 upon which a bearing 334 rides. The screw 332 is threadedly coupled to a traveler 336. The traveler 336 is fixedly coupled (either removably or permanently) to the carrier 314. The guide 342 and the screw 332 are coupled to an end cap 352 which is coupled to the spine 350 at an opposite end thereof from the actuator 318.

[0077] As the actuator 318 turns the screw via the coupler, the screw 332 rotates and the traveler 336 moves up and down on the screw 332 relative to the frame 320, moving the carrier 314 therewith. The motion of the two drive assemblies 308 is synchronized to reduce or prevent binding of the screws 332, misalignment of the carrier 314, etc.

[0078] The carrier 314 holds one or more (e.g., 96) vessels 312 and corresponding samples 110, 210. The high-throughput biological assay system 300 includes three zones, e.g., a first temperature control unit 302, a second temperature control unit 304, and a third temperature control unit 306. More or fewer temperature control units may be used as desired or a suitable for a given process. The first temperature control unit 302, second temperature control unit 304, and third temperature control unit 306 are separated from one another by spacers 346. The spacers 346 shown are of uniform height, but in other embodiments the spacers between two temperature control units (e.g., the first temperature control unit 302 and the second temperature control unit 304) may be of different size or height than the spacers 346 between two different temperature control units (e.g., the second temperature control unit 304 and the third temperature control unit 306). The temperature control units are supported on a frame 320 by standoffs 344. The frame 320 supports and ties the other components of the high-throughput biological assay system 300 together.

[0079] With reference to FIG. 3C and FIG. 3D, the carrier 314 includes a carrier body 322. The carrier body 322 may be a substantially planar body. One or more hubs 340 extends from a face (e.g., lower face) of the carrier body 322. In the example shown, the hubs 340 are luer hubs that receive a corresponding flange 328 of the vessels 312. The flange 328 is coupled and fluidically connected to a needle shaft 330. In the example shown, the hubs 340 are arranged in a standard 96-well format (e.g., arranged in an 8x12 array). The luer hubs 340 may be either “luer lock” hubs that threadedly receive the flange of a corresponding needle, or “luer slip” hubs that receive the flange with a slip-on fit. As shown for example in FIG. 3C and FIG. 3D, the hubs each have a receptacle 316 formed therein. The receptacle 316 is an aperture or passthrough that enables a fluid, such as air or a sample 110, 210 to pass through the carrier body 322 into the needle shaft 330 of the vessel 312.Docket No. G10083312P1PCT

[0080] Turning to FIG. 3E and FIG. 3F, an example of a first temperature control unit 302, second temperature control unit 304, or third temperature control unit 306 is shown. The temperature control units may have a body 324. The body 324 is substantially planar and of similar size and shape as the carrier 314. The body 324 includes a plurality of receptacles 310 therein, in a matching pattern and number to correspond to the hubs 340 of the carrier 314. FIG. 3E and FIG. 3F show an example of a vessel 312 needle shaft 330 received in one of the receptacles 310. In some embodiments, the receptacles 310 may be sized to provide a clearance to a needle shaft 330 disposed therein. In other embodiments, the receptacles 310 may be sized such that the needle shaft 330 touches a heater 412 in the zone. For example, the surface area of the vessel 208 in contact with the heater may be about 5-20 mm2. This relatively large contact area with respect to the volume of the sample 110, 210 enables rapid heating of the sample (e.g., up to about 2000°C / second. In many embodiments, the body 324 is, or includes a printed circuit board 400 as discussed further herein. See, e.g., FIG. 4A and FIG. 4B.

[0081] To assemble the high-throughput biological assay system 300, the components of the frame 320 may be coupled to one another, such as via one or more fasteners. The drive assemblies 308 may be assembled, such as by coupling the actuators 318 to their respective spines 350. The couplers 338 are attached to the actuator 318 shafts and to the screws 332 (e.g., at opposite ends of the couplers 338). The end cap 352 may be fitted over the ends of the screw 332 and the guide 342 and coupled to the spine 350.

[0082] The standoffs 344 may be coupled to the frame 320 and the desired zones (e.g., one or more of the first temperature control unit 302, the second temperature control unit 304, or the third temperature control unit 306). The first temperature control units may be coupled to, and separated from one another by, spacers 346. The carrier 314 is aligned with the zones such that the receptacles 316 align with the receptacles 310 to avoid binding of the needle shafts 330 as they pass through the temperature control units.

[0083] With reference to FIG. 4 A and FIG. 4B, a circuit layer 402 and a heater 412 of a printed circuit board 400 of one or more of the zones is shown. The circuit layer 402 includes a conductive path 404 such as may be formed by typical circuit printing methods. The conductive path 404 includes a first electrode 406 and a second electrode 408. The conductive path 404 may be formed of a material with a high electrical resistance that heats substantially (e.g., Joule heating) when an electrical current passes therethrough. For example, the conductive path 404 may be formed of copper, aluminum, a nickel-chrome alloy, or the like. Thus, the conductive path 404 may form a heater 412 when a source ofDocket No. G10083312P1PCT electricity is connected to the first electrode 406 and the second electrode 408. In some embodiments, the conductive path 404 may be between 2.5 and 10 meters long.

[0084] The conductive path 404 includes a plurality of clearances 410 therein. The clearances 410 correspond in pattern and number to the receptacles 310 which pass therethrough (not shown) and also to the receptacles 316 of the carrier 314. When the needle shafts 330 are received in the receptacles 310, the heater 412 imparts thermal energy to the sample 110, 210 in the vessels 312.

[0085] Turning to FIG. 4B, the circuit board 400 includes another circuit layer 414. The circuit layer 414 include a first coil 418 and a second coil 420 that form a sensor 424. The first coil 418 and second coil 420 may measure the temperature or resistivity of adjacent portions of the conductive path 404. Thus, a control system may be able to sense and adjust the electrical power delivered to the conductive path 404 to enable precise temperature control of the heater 412 and thus the samples 110, 210.

[0086] As with the circuit layer 402, the first coil 418 and second coil 420 include respective clearances 422 to enable the receptacles 310 and thus the needle shafts 330 to pass therethrough.

[0087] As shown, the first coil 418 and the second coil 420 may be duplicated and arranged in a sensor array 426 of many sensors 424.

[0088] In some embodiments, a circuit board includes photo sensors 424 such as photodiodes that can perform fluorescence detection, such as may be caused by a reaction taking place in the vessels 312. In some embodiments, the photo sensors 424 may be disposed above the carrier 314 or below the first temperature control unit 302, 304, and / or 306 and may detect light emitted by a sample from the tip 348 of the needle shaft 330 or via the receptacles 316.

[0089] FIG. 5 illustrates an example method 500 for operation of a high-throughput biological assay system such as the high-throughput biological assay system 100, the high- throughput biological assay system 200, or the high-throughput biological assay system 300. Although the example method 500 depicts a particular sequence of operations, the sequence may be altered without departing from the scope of the present disclosure. For example, some of the operations depicted may be performed in parallel or in a different sequence that does not materially affect the function of the method 500. In other examples, different components of an example device or system that implements the method 500 may perform functions at substantially the same time or in a specific sequence.Docket No. G10083312P1PCT

[0090] According to some examples, the method 500 includes providing a vessel containing a biological sample at operation 502. Operation of the high-throughput biological assay system 300 is substantially as described with respect to the high-throughput biological assay system 200. In the example of the high-throughput biological assay system 300, one or more vessels (e.g., hypodermic needles) are loaded with respective biological samples to be tested. The flanges 328 of the needles are coupled to the hubs 340 of the carrier 314 either before or after loading with the samples 110, 210. For example, the needles may be loaded with samples via the receptacles 316.

[0091] In operation 504, method 500 a temperature control unit is activated. For example, one or more of the first temperature control unit 302, the second temperature control unit 304, or the third temperature control unit 306 are activated to cause the conductive paths 404 therein to provide heat.

[0092] In operation 506, the control system 800 (see, e.g., FIG. 8) of the high-throughput biological assay system moves the vessel, via the actuators or drive assemblies 308, proximate to the temperature control unit to control a temperature of the biological sample within the vessel. As shown for example in FIG. 3G, the carrier 314 is shown with the needle shafts 330 penetrating the receptacles 310 of the first temperature control unit 302, passing through the first temperature control unit 302 body 324 and just beginning to enter the receptacles 310 of the second temperature control unit 304. For simplicity and clarity, only some of the hubs 340 are populated with vessels 312. However, in practice, any number of the hubs 340 may receive vessels 312 and samples 110, 210.

[0093] In the position shown for example in FIG. 3G, the needle shafts 330 of the vessels 312 are heated until the samples therein achieve a desired temperature. The carrier 314 may be held in this position for a sufficient time for the samples 110, 210 to reach a desired temperature and to enable a desired reaction to take place. After the desired temperature and dwell time has been achieved, the drive assemblies 308 may move the carrier 314 to a different position with respect to the zones.

[0094] In operation 506 of method 500 a second temperature control unit is activated. For example, one or more of the first temperature control unit 302, the second temperature control unit 304, or the third temperature control unit 306 are activated to cause the conductive paths 404 therein to provide heat. In many embodiments of the method 500, the operation 504 and the operation 506 may be executed at the same time. In some embodiments, the operation 504 and the operation 506 may be executed in series, with the first temperature control unit being de-activated before activating the second temperatureDocket No. G10083312P1PCT control unit, such as to reduce power draw of the high-throughput biological assay system 300.

[0095] According to some examples, the method 500 includes moving the vessel proximate to the second temperature control unit to control a second temperature of the biological sample within the vessel at operation 510. For example, turning to FIG. 3H, the carrier 314 is shown in a lowered position relative to its position in FIG. 3G. Here, the tips 348 of the needle shafts 330 are just penetrating below the lower face of the body 324 of the third temperature control unit 306. FIG. 3G and FIG. 3H are example positions, only. The drive assemblies 308 may, via synchronized turning of their respective actuators 318 and screws 332 move the carrier 314 anywhere along the screw relative to the first temperature control units 302, 304, and / or 306. Thus, for example, the high-throughput biological assay system 300 can execute the PCR process or another process as described with respect to the high- throughput biological assay system 200.

[0096] In other embodiments, one or more of the first temperature control units 302, second temperature control unit 304, or third temperature control unit 306 are coupled to the drive assemblies 308 and are moveable thereby, while the carrier 314 holding the vessels 312 and samples 110, 210 is stationary relative to the frame 320.

[0097] Referring to FIG. 6, a high-throughput biological assay system 600 represents an alternative embodiment that uses electrostatic forces to move aqueous samples across capacitive pads on a printed circuit board 602, rather than mechanical movement of vessels. The high-throughput biological assay system 600 may provide faster throughput and higher rates of heating and cooling compared to the high-throughput biological assay system 200. The sample 110 may be moved electrostatically through the first zone 202, the second zone 204, and the third zone 206 to complete respective portions of a PCR reaction without physical movement of vessels or the actuator 102.

[0098] The printed circuit board 602 includes multiple capacitive pads 604a, 604b, 604c, and 604d arranged to enable controlled movement of aqueous samples. The capacitive pads generate electrostatic forces that attract and move water droplets containing biological samples across the surface, taking advantage of the polar nature of water molecules for precise control over sample positioning and movement between different temperature zones.

[0099] Heaters 606a, 606b, and 606c are embedded within the printed circuit board 602 as copper traces to provide localized temperature control for each zone. Each heater corresponds to a specific temperature zone, with heater 606a associated with the first zone 202, heaterDocket No. G10083312P1PCT 606b with the second zone 204, and heater 606c with the third zone 206, providing more precise temperature control compared to external heating elements.

[0100] Temperature sensors 610a, 610b, and 610c are positioned adjacent to the respective heaters to monitor and control the temperature of each zone. The sensors may be implemented as copper traces functioning as resistance temperature detectors (RTDs), thermistors, thermocouples, or integrated circuit temperature sensors. For RTD implementations, the resistance of the copper changes as a function of temperature and may be coupled to reference resistors to enable precise temperature measurement. The sensors are integrated into a closed- loop control system, such as a proportional-integral-derivative (PID) control loop, where the processing element 802 receives temperature data and executes control algorithms to adjust the electrical power and / or duty cycle supplied to the respective heaters to achieve and maintain desired temperature setpoints.

[0101] In some embodiments, the heaters 606a, 606b, and 606c and the sensors 610a, 610b, and 610c may be integrated as respective single devices. The resistance across the conductive path of the heater can be measured and used to provide temperature measurement, as the electrical resistance varies predictably with temperature changes. This dual-function approach allows the conductive path heater to simultaneously provide both heating capability and temperature sensing functionality. The temperature coefficient of resistance for the material forming the heater may be calibrated to provide accurate temperature readings, reducing circuit board complexity by eliminating the need for separate temperature sensing elements while maintaining precise temperature control capabilities.

[0102] A dielectric layer 608 is positioned over the capacitive pads 604a, 604b, 604c, and 604d to separate the pads from the aqueous sample 110. The dielectric layer 608 has a thickness of approximately 10 microns and is formed from a transparent perfluorinated material such as ethylene tetrafluoroethylene (ETFE), enabling the application of alternating voltage to the capacitive pads while maintaining electrical isolation from the sample 110.

[0103] A cap layer 704 is positioned above the dielectric layer 608 to prevent evaporation of aqueous samples during processing. In some embodiments, the cap layer 704 is made of glass or another suitable material and maintains sample integrity by preventing or reducing evaporation of the sample 110 while allowing optical measurements to be performed. The cap layer 704 is spaced apart from the dielectric layer 608 suitably far to enable the sample 110 to be moved about the printed circuit board 602 by the capacitive pads 604.

[0104] The printed circuit board 602 includes apertures 612 positioned to allow optical access to the sample 110 during processing. The apertures may be located through the heatersDocket No. G10083312P1PCT and / or through the capacitive pads. Fluorescence sensors 614 are positioned adjacent to the apertures 612 to detect fluorescence signals generated by the sample 110 in response to completion of chemical reactions. The apertures 612 and the transparent nature of the dielectric layer 608 enable light to pass through the entire structure for optical detection and analysis. For example, a fluorescence sensor 614 is positioned adjacent to the aperture 612 to detect optical signals generated by biological samples and is configured to detect doublestrand DNA or primer fluorescence through the transparent aperture 612, enabling real-time monitoring of PCR reactions and other biological processes occurring within the sample 110.

[0105] Referring to FIG. 7A, the high-throughput biological assay system 600 may include a sample source 702 configured to supply the sample 110 for electrostatic transfer to the capacitive pad 604a. The sample source 702 may include a reservoir that holds the sample 110 through surface tension containment, allowing the sample 110 to be maintained in position until electrostatic forces are applied. The sample source 702 may further include a sponge and / or a wick positioned near the capacitive pad 604a to facilitate transfer of the sample 110 from the reservoir to the dielectric layer 608.

[0106] As shown in FIG. 7A, the capacitive pad 604a may be configured to apply an alternating voltage that creates electrostatic attraction forces acting on the sample 110. The alternating voltage applied to the capacitive pad 604a may utilize the polar character of water molecules within the sample 110 to generate attractive forces that pull the sample 110 from the sample source 702 toward the capacitive pad 604a. The polar nature of water molecules may cause the sample 110 to respond to the electric field generated by the capacitive pad 604a, enabling controlled movement of the sample 110 across the dielectric layer 608 and to the first zone 202.

[0107] The electrostatic attraction created by the capacitive pad 604a may overcome the surface tension forces holding the sample 110 within the sample source 702, allowing the sample 110 to be drawn from the reservoir and transferred onto the dielectric layer 608. The wick component of the sample source 702 may facilitate this transfer by providing a pathway for the sample 110 to move from the reservoir toward the electrostatic field generated by the capacitive pad 604a. Once positioned over the capacitive pad 604a, the sample 110 may be further manipulated through application of alternating voltages to adjacent capacitive pads to move the sample 110 between different temperature zones for PCR processing.

[0108] Referring to FIG. 7B, the sample 110 may be moved to the first zone 202 through electrostatic attraction generated by electrical excitation of the capacitive pad 604a and the capacitive pad 604b. The capacitive pad 604a and the capacitive pad 604b may be configuredDocket No. G10083312P1PCT to apply alternating voltages that create electrostatic forces acting on the polar water molecules within the sample 110. The electrostatic forces generated by the capacitive pad 604a and the capacitive pad 604b may pull the sample 110 from the sample source 702 toward the first zone 202, where the sample 110 may be positioned for thermal processing.

[0109] In the first zone 202, the heater 606a may be activated to heat the sample 110 to a temperature range suitable for the denaturation step of a PCR process. The heater 606a may maintain the temperature of the sample 110 in the range of 94°C to 98°C, which may be suitable for performing a denaturation operation. The temperature sensor 610a may monitor the temperature conditions within the first zone 202 to provide feedback for precise temperature control during the denaturation process.

[0110] During the denaturation operation in the first zone 202, the elevated temperature maintained by the heater 606a may cause double-stranded DNA within the sample 110 to separate into single strands. The thermal energy provided by the heater 606a may break the hydrogen bonds holding the two DNA strands together, resulting in the formation of singlestranded DNA templates. The separation of double-stranded DNA into single strands may enable each strand to serve as a template for subsequent synthesis steps in the PCR process.

[0111] The electrostatic positioning system using the capacitive pad 604a and the capacitive pad 604b may enable rapid movement of the sample 110 to the first zone 202, while the embedded heater 606a may provide faster heating rates compared to external heating elements. The combination of electrostatic sample movement and embedded heating elements may enable the high-throughput biological assay system 600 to achieve higher rates of heating and cooling during the denaturation step, thereby providing faster throughput for PCR processing compared to mechanical vessel movement systems.

[0112] Referring to FIG. 7C, the sample 110 may be moved to the second zone 204 through electrostatic attraction generated by electrical excitation of the capacitive pad 604b and the capacitive pad 604c. The capacitive pad 604b and the capacitive pad 604c may be configured to apply alternating voltages that create electrostatic forces acting on the polar water molecules within the sample 110. The electrostatic forces generated by the capacitive pad 604b and the capacitive pad 604c may pull the sample 110 from the first zone 202 toward the second zone 204, where the sample 110 may be positioned for the annealing step of the PCR process.

[0113] In the second zone 204, the heater 606b may be activated to control the temperature of the sample 110 to a range suitable for the annealing operation of a PCR process. The heater 606b may maintain the temperature of the sample 110 in the range of 50°C to 65°C, whichDocket No. G10083312P1PCT may be suitable for performing an annealing operation. The temperature sensor 610b may monitor the temperature conditions within the second zone 204 to provide feedback for precise temperature control during the annealing process.

[0114] During the annealing operation in the second zone 204, the controlled temperature maintained by the heater 606b may enable primers to bind to their complementary sequences on the single-stranded DNA templates created during the denaturation step. The temperature range of 50°C to 65°C provided by the heater 606b may allow the primers to form hydrogen bonds with their complementary sequences on the single-stranded DNA templates. The primers may be short single-stranded sequences of nucleotides designed to match regions flanking the target DNA sequence, and the annealing process may enable the primers to attach to the appropriate locations on the template DNA.

[0115] The electrostatic positioning system using the capacitive pad 604b and the capacitive pad 604c may enable rapid movement of the sample 110 to the second zone 204, while the embedded heater 606b may provide precise temperature control for the annealing step. The combination of electrostatic sample movement and embedded heating elements may enable the high-throughput biological assay system 600 to achieve controlled temperature transitions during the annealing step, thereby providing faster throughput for PCR processing compared to mechanical vessel movement systems.

[0116] Referring to FIG. 7D, the sample 110 may be moved to the third zone 206 through electrostatic attraction generated by electrical excitation of the capacitive pad 604c and the capacitive pad 604d. The capacitive pad 604c and the capacitive pad 604d may be configured to apply alternating voltages that create electrostatic forces acting on the polar water molecules within the sample 110. The electrostatic forces generated by the capacitive pad 604c and the capacitive pad 604d may pull the sample 110 from the second zone 204 toward the third zone 206, where the sample 110 may be positioned for the extension step of the PCR process.

[0117] In the third zone 206, the heater 606c may be activated to control the temperature of the sample 110 to a range suitable for the extension operation of a PCR process. The heater 606c may maintain the temperature of the sample 110 in the range of 72°C to 75°C, which may be suitable for performing an extension operation. The temperature sensor 610c may monitor the temperature conditions within the third zone 206 to provide feedback for precise temperature control during the extension process.

[0118] During the extension operation in the third zone 206, the controlled temperature maintained by the heater 606c may enable DNA polymerase to synthesize a new strand ofDocket No. G10083312P1PCT DNA complementary to the DNA template strands. The DNA polymerase may attach to the primers bound to the single-stranded DNA templates from the annealing step and may extend the primers by adding nucleotides to the end of each primer. The extension process may result in the synthesis of new DNA strands complementary to the template strands, leading to the formation of new double-stranded DNA starting from the primers.

[0119] The high-throughput biological assay system 600 may be configured to cycle the sample 110 through the first zone 202, the second zone 204, and the third zone 206 repeatedly to achieve a desired level of DNA amplification. The electrostatic forces generated by the capacitive pads 604a, 604b, 604c, and 604d may enable the sample 110 to be moved sequentially through each temperature zone multiple times, with each cycle comprising denaturation in the first zone 202, annealing in the second zone 204, and extension in the third zone 206. The cycling process may be repeated tens to hundreds of times to achieve sufficient amplification of target DNA sequences within the sample 110.

[0120] Between processing of different samples, the high-throughput biological assay system 600 may perform a cleaning process to remove residual material from the dielectric layer 608. A washing solution may be manipulated around the surface of the dielectric layer 608 using the same electrostatic forces generated by the capacitive pads 604a, 604b, 604c, and 604d. The washing solution may be moved across the surface to remove any remaining biological material, reagents, or contaminants from previous samples, thereby preparing the system for processing of subsequent samples. The cleaning process may help prevent crosscontamination between different samples and may maintain the accuracy and reliability of the biological assay results.

[0121] Although the examples illustrated in FIGS. 6-7D show three temperature zones and four capacitive pads arranged in a linear fashion, it should be understood that other embodiments may have different configurations. For example, other embodiments may include more or fewer temperature zones, such as one, two, four, or more zones depending on the specific biological assay requirements. Similarly, the number of capacitive pads may vary, with embodiments having two, three, five, or more capacitive pads as needed for sample manipulation. The arrangement of the capacitive pads and temperature zones may also differ from the linear configuration shown, with alternative embodiments featuring square or rectangular two-dimensional arrays, circular arrangements, loop configurations, or other geometric patterns suitable for the intended application.

[0122] FIG. 8 is a simplified block diagram of components of a control system 800 of the high-throughput biological assay system 100, the high-throughput biological assay systemDocket No. G10083312P1PCT200, or the high-throughput biological assay system 300. For example, the processing element 802 and the memory component 808 may be located at one or in several control systems 800. This disclosure contemplates any suitable number of such control systems 800. For example, the control system 800 may be a desktop computing system, a mainframe, a blade, a mesh of control systems 800, a laptop or notebook control system 800, a tablet control system 800, an embedded control system 800, a system-on-chip, a single-board control system 800, or a combination of two or more of these. Where appropriate, a control system 800 may include one or more control systems 800; be unitary or distributed; span multiple locations; span multiple machines; span multiple data centers; or reside in a cloud, which may include one or more cloud components in one or more networks. A control system 800 may include one or more processing elements 802, an input / output I / O interface 804, one or more external devices 812, one or more memory components 608, and a network interface 810. Each of the various components may be in communication with one another through one or more buses or communication networks, such as wired or wireless networks, e.g., a network. The components in FIG. 8 are exemplary only. In various examples, the control system 800 may include additional components and / or functionality not shown in FIG. 8.

[0123] The processing element 802 may be any type of electronic device capable of processing, receiving, and / or transmitting instructions. For example, the processing element 802 may be a central processing unit, microprocessor, processor, or microcontroller. Additionally, it should be noted that some components of the control system 800 may be controlled by a first processing element 802 and other components may be controlled by a second processing element 802, where the first and second processing elements may or may not be in communication with each other.

[0124] The I / O interface 804 allows a user to enter data in to control system 800, as well as provides an input / output for the control system 800 to communicate with other devices or services. The I / O interface 804 can include one or more input buttons, touch pads, touch screens, and so on.

[0125] The external device 812 are one or more devices that can be used to provide various inputs to the computing systems 600, e.g., mouse, microphone, keyboard, trackpad, sensing element (e.g., a thermistor, humidity sensor, light detector, etc. The external devices 812 may be local or remote and may vary as desired. In some examples, the external devices 812 may also include one or more additional sensors.Docket No. G10083312P1PCT

[0126] The memory components 808 are used by the control system 800 to store instructions for the processing element 802 such as the operations of the PCR process, setpoints for the temperatures of the zones, control algorithms for the sensors to enable control of the heater 412, or data captured from the photo sensors, etc. The memory components 808 may be, for example, magneto-optical storage, read-only memory, random access memory, erasable programmable memory, flash memory, or a combination of one or more types of memory components.

[0127] The network interface 810 provides communication to and from the control system 800 to other devices. The network interface 810 includes one or more communication protocols, such as, but not limited to Wi-Fi, Ethernet, Bluetooth, etc. The network interface 810 may also include one or more hardwired components, such as a Universal Serial Bus (USB) cable, or the like. The configuration of the network interface 810 depends on the types of communication desired and may be modified to communicate via Wi-Fi, Bluetooth, etc.

[0128] The display 806 provides a visual output for the control system 800 and may be varied as needed based on the device. The display 806 may be configured to provide visual feedback to a user and may include a liquid crystal display screen, light emitting diode screen, plasma screen, or the like. In some examples, the display 806 may be configured to act as an input element for the user through touch feedback or the like.

[0129] The description of certain embodiments included herein is merely exemplary in nature and is in no way intended to limit the scope of the disclosure or its applications or uses. In the included detailed description of embodiments of the present systems and methods, reference is made to the accompanying drawings which form a part hereof, and which are shown by way of illustration specific to embodiments in which the described systems and methods may be practiced. These embodiments are described in sufficient detail to enable those skilled in the art to practice presently disclosed systems and methods, and it is to be understood that other embodiments may be utilized, and that structural and logical changes may be made without departing from the spirit and scope of the disclosure. Moreover, for the purpose of clarity, detailed descriptions of certain features will not be discussed when they would be apparent to those with skill in the art so as not to obscure the description of embodiments of the disclosure. The included detailed description is therefore not to be taken in a limiting sense, and the scope of the disclosure is defined only by the appended claims.Docket No. G10083312P1PCT

[0130] From the foregoing it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention.

[0131] The particulars shown herein are by way of example and for purposes of illustrative discussion of the preferred embodiments of the present disclosure and are presented in the cause of providing what is believed to be the most useful and readily understood description of the principles and conceptual aspects of various embodiments of the invention. In this regard, no attempt is made to show structural details of the invention in more detail than is necessary for the fundamental understanding of the invention, the description taken with the drawings and / or examples making apparent to those skilled in the art how the several forms of the invention may be embodied in practice.

[0132] As used herein and unless otherwise indicated, the terms “a” and “an” are taken to mean “one”, “at least one” or “one or more”. Unless otherwise required by context, singular terms used herein shall include pluralities and plural terms shall include the singular.

[0133] Unless the context clearly requires otherwise, throughout the description and the claims, the words ‘comprise’, ‘comprising’, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”. Words using the singular or plural number also include the plural and singular number, respectively. Additionally, the words “herein,” “above,” and “below” and words of similar import, when used in this application, shall refer to this application as a whole and not to any particular portions of the application.

[0134] All relative, directional, and ordinal references (including top, bottom, side, front, rear, first, second, third, and so forth) are given by way of example to aid the reader’s understanding of the examples described herein. They should not be read to be requirements or limitations, particularly as to the position, orientation, or use unless specifically set forth in the claims. Connection references (e.g., attached, coupled, connected, joined, and the like) are to be construed broadly and may include intermediate members between a connection of elements and relative movement between elements. As such, connection references do not necessarily infer that two elements are directly connected and in fixed relation to each other, unless specifically set forth in the claims.

[0135] Of course, it is to be appreciated that any one of the examples, embodiments or processes described herein may be combined with one or more other examples, embodiments and / or processes or be separated and / or performed amongst separate devices or device portions in accordance with the present systems, devices and methods.Docket No. G10083312P1PCT

[0136] Finally, the above discussion is intended to be merely illustrative of the present system and should not be construed as limiting the appended claims to any particular embodiment or group of embodiments. Thus, while the present system has been described in particular detail with reference to exemplary embodiments, it should also be appreciated that numerous modifications and alternative embodiments may be devised by those having ordinary skill in the art without departing from the broader and intended spirit and scope of the present system as set forth in the claims that follow. Accordingly, the specification and drawings are to be regarded in an illustrative manner and are not intended to limit the scope of the appended claims.

Claims

Docket No. G10083312P1PCTCLAIMSWhat is claimed is:

1. A biological assay system comprising: a vessel configured to receive a biological sample; a temperature control unit; and a carrier configured to move the vessel proximate to the temperature control unit to control a temperature of the biological sample within the vessel.

2. The system of claim 1, wherein the vessel comprises a thin-walled cylindrical element.

3. The system of claim 2, wherein a wall thickness of the vessel ranges from 0.10 mm to 0.35 mm.

4. The system of claim 1, wherein the biological sample has a volume of less than 0.5 pL.

5. The system of claim 1, wherein the vessel has a surface area of contact with the temperature control unit of between 5 and 20 mm2.

6. The system of claim 1, wherein the vessel comprises a hypodermic needle.

7. The system of claim 1, wherein the actuator comprises a stepper motor.

8. The system of claim 1, further comprising a second temperature control unit and a third temperature control unit, wherein the actuator moves the vessel between the temperature control units to perform a polymerase chain reaction (PCR) process.

9. The system of claim 1, wherein the temperature control unit is configured to achieve a temperature change rate in the sample up to 2000 °C per second.

10. The system of claim 1, wherein the carrier is configured to couple to a plurality of vessels and move each of the plurality of vessels proximate to the temperature control unit.

11. The system of claim 1, further comprising a light sensor configured to detect fluorescence generated by the biological sample in response to completion of a chemical reaction.Docket No. G10083312P1PCT12. A biological assay method comprising: providing a vessel containing a biological sample; providing a temperature control unit; moving the vessel proximate to the temperature control unit to control a temperature of the biological sample within the vessel.

13. The biological assay method of claim 12, further comprising performing an operation of a PCR process in the vessel at the temperature.

14. The method of claim 13, wherein the operation comprises a denaturation operation and the temperature control unit maintains the temperature in the range of 94 °C to 98 °C.

15. The method of claim 13, wherein the operation comprises an annealing operation and the temperature control unit maintains the temperature in the range 50 °C to 65 °C.

16. The method of claim 13, wherein the operation comprises an extension operation and the temperature control unit maintains the temperature in the range 72 °C to 75 °C.

17. The method of claim 13, wherein the biological sample has a volume of less than 0.5 pL.

18. The method of claim 13, wherein the vessel has a surface area of contact with the temperature control unit of between 5 and 20 mm2.

19. The method of claim 13, further comprising moving the vessel proximate to a second temperature control unit and a third temperature control unit, wherein the actuator moves the vessel between the temperature control units to perform a polymerase chain reaction (PCR) process.

20. A biological assay system comprising: a plurality of needles configured to receive a respective plurality of biological samples; a plurality of temperature control units; and a carrier configured to receive the plurality of needles and to move the plurality of needles proximate to the plurality of temperature control units, such that each temperature control unit of the plurality of temperature control units controls a temperature of the plurality of biological samples.

21. A high-throughput biological assay system comprising:Docket No. G10083312P1PCT a printed circuit board; a plurality of capacitive pads disposed on the printed circuit board and configured to generate electrostatic forces to move an aqueous sample containing a biological sample across a surface of the printed circuit board; a plurality of heaters embedded within the printed circuit board, each heater corresponding to a temperature zone; and a dielectric layer positioned over the capacitive pads and configured to separate the capacitive pads from the aqueous sample while enabling the electrostatic forces to act on the aqueous sample.

22. The system of claim 21, further comprising a plurality of temperature sensors positioned adjacent to the respective heaters to monitor temperature conditions within each temperature zone.

23. The system of claim 21, wherein the plurality of capacitive pads comprises a first capacitive pad, a second capacitive pad, a third capacitive pad, and a fourth capacitive pad arranged to enable sequential movement of the aqueous sample between temperature zones.

24. The system of claim 21, further comprising a sample source configured to supply the aqueous sample for electrostatic transfer to the capacitive pads.

25. The system of claim 21, wherein the plurality of heaters comprises a first heater configured to maintain a temperature suitable for denaturation, a second heater configured to maintain a temperature suitable for annealing, and a third heater configured to maintain a temperature suitable for extension.

26. The system of claim 21, further comprising at least one aperture extending through the printed circuit board to allow optical access to the aqueous sample.

27. The system of claim 26, further comprising a fluorescence sensor positioned adjacent to the aperture to detect fluorescence signals generated by the biological sample.

28. The system of claim 21, wherein the dielectric layer has a thickness of approximately 10 micrometers and is formed from a transparent fluorinated material.

29. The system of claim 21, further comprising a cap layer positioned above the dielectric layer to prevent evaporation of the aqueous sample during processing.Docket No. G10083312P1PCT30. The system of claim 21, wherein the capacitive pads are configured to apply alternating voltages that utilize polar characteristics of water molecules within the aqueous sample to generate attractive forces for sample movement.

31. The system of claim 21, wherein the system is configured to perform a polymerase chain reaction process by moving the aqueous sample sequentially through a first zone for denaturation, a second zone for annealing, and a third zone for extension.

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