Fluid detection and control algorithms for PCR analysis

Abstract

Various embodiments of systems and methods for detecting and real-time adjusting the position control of fluid volume moving through an optical detection region in a fluid channel of a chip on an RT-qPCR system are disclosed. These embodiments are disclosed within an RT-qPCR system having a chip and a housing for receiving the chip, as well as further mechanical means for interacting with a membrane of the chip for position control. Furthermore, exemplary embodiments include systems and methods for capacitance detection of fluid within a fluid channel and optical detection within an optical detection region of the fluid channel within the chip.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 093,640, filed October 19, 2020, entitled “Collection Point qPCR System,” pursuant to PCT Section 8. This application also relates to PCT applications entitled “Methods and Apparatus for Controlling Fluid Volume to Achieve Separation and PCR Amplification,” “Disposable Casing for Reagent Storage and Method of Use Thereof,” and “Apparatus with Fluid Channel Geometry for Sample-to-Result PCR Analysis and Method of Use Thereof,” as well as U.S. Design Application No. 29 / 812,034, entitled “Fluid Channel Geometry of a Chip,” all filed simultaneously on October 19, 2021, and all listing the same applicant, Formulatrix, Inc. The contents of the foregoing applications are incorporated herein by reference in their entirety as if fully set forth herein. Technical Field

[0003] This invention relates to real-time polymerase chain reaction (qPCR), systems, and methods, namely, functional PCR systems and methods for improving qPCR processing and analysis through optical and electrical detection of samples. Background Technology

[0004] Real-time polymerase chain reaction (real-time PCR), also known as quantitative PCR or (qPCR), is a technique that monitors DNA amplification in real time, rather than at the end of the polymerase chain reaction procedure, for quantitative analysis. There are two common methods for real-time detection of PCR products. The first is using non-specific fluorescent dyes to insert the DNA, and the second is using sequence-specific DNA probes, which consist of oligonucleotides labeled with fluorescent reporters, allowing detection only after the probe hybridizes to its complementary sequence.

[0005] Reverse transcription polymerase chain reaction (RT-PCR) is a technique that combines reverse transcription of RNA into DNA, complementary DNA (also known as cDNA), and amplification of specific DNA targets. By combining RT-PCR with qPCR (RT-qPCR), technicians can analyze and quantify viral RNA gene expression.

[0006] Historically, performing RT-qPCR has required skilled technicians and complex, expensive equipment. The disclosures in this paper provide improvements to laboratory techniques and methods for performing RT-qPCR by adding mechanical, computational, and biological functions to enhance quantitative analysis capabilities. Furthermore, systems and methods for error verification, fluid detection, and tracking allow for improvements in consumable-based chip analysis by controlling error detection, tracking, and motor functions, as well as sample velocity and sample integrity. Summary of the Invention

[0007] This paper discloses a system and method for detecting and real-time adjusting the position control of fluid volume. The fluid volume can be located within a membrane-equipped chip, where RT-qPCR is cyclically performed.

[0008] In some embodiments, for example, a system is disclosed for detecting and real-time adjusting the positional control of a fluid volume moving through an optical detection region in a fluid channel. The system includes mechanical means. It is also configured with a chip including a fluid channel, wherein the mechanical means control the movement of the fluid volume. Finally, the chip further includes an optical detection region that may contain voxels. The system is also equipped with one or more independent heating blocks. The chip is at least partially positioned on one or more independent heating blocks such that the fluid channel is adjacent to one or more independent heating blocks. Furthermore, the system is equipped with an optical detection unit including optical light-emitting elements, sometimes including three light-emitting diodes and a dual-band filter. The optical detection unit of the system also includes an optical detector and a processing unit, the optical detector may be multiple detection diodes, and the processing unit is used to analyze the optical detection region of the chip.

[0009] In an additional embodiment, a method for detecting and real-time adjusting the positional control of fluid volume moving through an optical detection region in a fluid channel is disclosed. The method includes preparing a sample with a fluorescent marker. Next, a chip is configured to receive the sample, wherein the chip has a fluid channel for sample flow and an optical detection region allowing optical light to be transmitted to the fluid channel. Next, the sample is applied to the chip, and the chip is applied to two heating arrays. Then, an optical detection unit is activated, wherein the activation optical detection unit is configured to illuminate one or more LEDs and activate optical detection diodes. Next, a mechanical device is activated to press down a region of the chip, wherein the pressed region causes fluid within the chip, including the sample, to move along the fluid channel and through the two heating arrays. The presence of the sample within the optical detection region and a signal output of the sample are then detected by the optical detection unit. The method then adjusts a motion control script based at least on the signal output, wherein the motion control script at least adjusts the initiation and / or stopping of the mechanical device.

[0010] In a further embodiment, a method for detecting and real-time adjusting the positional control of fluid volume moving into a fluid channel is disclosed. The method includes preparing a sample using magnetic beads adsorbing nucleic acid compounds. Next, a chip is configured to receive the sample, wherein the chip has fluid channels for sample flow, and the fluid channels pass through two separate heating arrays. Next, the sample is applied to the chip, and a capacitance array is applied along a section of the fluid channel of the chip (also referred to as a capacitance sensing region). Next, a mechanical device is activated to press down a region of the chip, wherein the pressed region causes fluid within the chip, including the sample, to move along the fluid channels and through the two separate heating arrays. The method then detects the capacitance change in the fluid channel at the capacitance array using a capacitance sensor within the capacitance sensing region. Attached Figure Description

[0011] Many aspects of this disclosure can be better understood with reference to the following accompanying drawings. The components in the drawings are not necessarily to scale, but are intended to clearly illustrate the principles of this disclosure. Furthermore, in the drawings, the same reference numerals denote corresponding parts in several views. In the drawings:

[0012] Figure 1 This is a diagram of an exemplary embodiment of a fluid RT-qPCR system for RT-qPCR analysis;

[0013] Figure 2 This is for fluid RT-qPCR systems Figure 1 An illustration of an exemplary embodiment with the outer casing removed;

[0014] Figure 3 This is a diagram illustrating an exemplary embodiment of an optical detection unit;

[0015] Figure 4A -E is an illustration of an exemplary embodiment of a sample moving through an optical detection region via a fluid channel, wherein the analog-to-digital converter (ADC) count reflects the position;

[0016] Figure 5 This is a diagram illustrating an exemplary implementation of the chip;

[0017] Figure 6 This is an illustration of an exemplary embodiment of a capacitive sensing array used with a chip;

[0018] Figure 7 This is a diagram of an exemplary implementation of farads detected as a sample moves across a fluid channel on a capacitive sensing array;

[0019] Figure 8 This is a diagram illustrating an exemplary embodiment of a capacitance signal;

[0020] Figure 9This is a diagram of an exemplary implementation of conventional cyclic RT-qPCR amplification time;

[0021] Figure 10 This is a diagram of an exemplary implementation of the function disclosed herein in improving the timing of RT-qPCR amplification. Detailed Implementation

[0022] In the following detailed description, reference is made to the accompanying drawings, which form a part thereof. In the drawings, similar symbols generally identify similar components unless the context otherwise requires. The illustrative embodiments described in the detailed description, drawings, and claims are not intended to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the scope of the subject matter set forth herein. It will be readily understood that aspects of this disclosure, as illustrated in the general description and drawings herein, can be arranged, substituted, combined, separated, and designed in a variety of different configurations, all of which are expressly contemplated herein.

[0023] Unless otherwise expressly stated, disjunctive language such as the phrase “at least one of X, Y or Z” can in other respects be any one of X, Y, Z or any combination thereof (e.g., X, Y and / or Z) depending on the context in which it is typically used to indicate terms, terms, etc. Therefore, such disjunctive language is generally not intended and should not imply that certain embodiments require at least one of X, at least one of Y, or at least one of Z, each of which must be present.

[0024] Now for reference Figure 1 An exemplary embodiment of a fluid RT-qPCR system 100 is disclosed. A housing 102 protects internal components, including mechanical means for pushing pins into a membrane of a chip via a cartridge, and processing equipment. A display unit or screen for displaying a graphical user interface to a technician utilizing the fluid RT-qPCR system 100 constructed within the housing 102 is not described. A cartridge or consumable cartridge slides into a cartridge insert 104 within the housing. The chip is configured within the cartridge, wherein a sample and / or fluid is first placed on the chip, and then the chip is inserted into the cartridge insert 104. The system is sized in the range of 12 inches to 36 inches in height and width and can be configured to fit existing laboratory environments by positioning the internal hardware components in different configuration arrays. For the purposes of this disclosure, samples and fluids are interchangeable.

[0025] As discussed, the qPCR system is a real-time qPCR system (RT-qPCR) in which the sample is observed in real time during the amplification process. The fluid RT-qPCR system 100 is designed to handle many tasks that might traditionally be performed by technicians. The chip used to insert into the cartridge is a consumable, allowing for single-use and disposal in accordance with various regulations and / or protocols.

[0026] Now for reference Figure 2 This refers to fluid RT-qPCR systems used for RT-qPCR analysis. Figure 1 The illustration shows an exemplary embodiment with the casing removed. In this example, a mechanical device 202 moves about a central axis via a track. This device is equipped with nodes or ridges that, as they travel around the track, strike a set of pins that press against a rubber diaphragm on the chip, thereby allowing fluid trapped within the chip to circulate to different areas. These different areas include heating areas, magnet areas, and capacitor array areas. Figure 5 Further detailed descriptions of various regions of an exemplary chip are disclosed herein. The mechanical device may be a top roller, drum, or cam, which is inherently rigid, having a smooth exterior and nodules for pressing pins onto a surface film of the chip. The top roller or cam may be composed of metal or a hard polymer; furthermore, a motor configured to the mechanical device allows for starting and stopping, as well as changing the acceleration and speed of the drum. The motor is connected via a shaft or gearbox to drive the top roller or cam. Input received from the optical detection unit and / or capacitor array allows for the configuration of control algorithms for the mechanical device 202.

[0027] In some instances, an algorithm is defined within the logic of the processing unit, where a specific measurement of the analog-to-digital converter (ADC) count on the fluorescent marker can send a feedback loop to stop the mechanism after a specified time, such as 10 seconds after passing through the optical detection area. Similarly, the mechanism can be triggered to begin rotating around a central axis when the ADC count has cleared the voxel or optical detection area and has spent a specified time period on the heating unit. For the purposes of this disclosure, heating unit, heating block, heating zone, or heating element refers to the same thing, and in some instances, multiple heating elements, each having a varying temperature range, may be part of the implementation.

[0028] Continuing, the consumable cartridge, which can also be disposed of or reused, is placed into the consumable cartridge insert 204 within the system, wherein the cartridge is aligned with various pins oriented below the mechanical device 202. When the motor 210 engages, in response to a predefined function of the processing unit or system, or as part of initial system parameters, the mechanical device 202 rotates about a central axis, pressing the pins to circulate fluid within the chip's fluid channels. A sample is placed within the chip, which is configured with multiple washing solutions and areas for disposing of waste liquid.

[0029] The processing unit can be mounted on a single PCB board and can have multiple components, such as a GPU, RAM, SSD, as well as peripherals and I / O for additional input and output to the peripherals. It is conceivable that the system described in this paper can communicate via data cable or wireless protocol and can be adapted and configured for a cloud environment, where a cluster of devices can form a system capable of performing diagnostics and tests on multiple devices.

[0030] In one instance, the sample can be prepared using a fluorescent label (dye), such as FAM. TM HEX TM ROX TM TET TM JOE TM VIC TM NED TM PET TM , TAMRA TM Or any other fluorescent dye used for DNA sequencing. Many of these dyes are excited at a single wavelength of 488 nm, but emit at distinctly different wavelengths. The table below provides examples of various fluorescent dyes that can be combined with those disclosed herein.

[0031] Table 1. Examples of fluorescent dyes

[0032] name <![CDATA[λ max / nm (absorption) <![CDATA[λ max / nm(emission)]]> <![CDATA[FAM TM ]]> 494 518 <![CDATA[HEX TM ]]> 535 556 <![CDATA[ROX TM ]]> 575 602 <![CDATA[TAMRA TM ]]> 555 580 <![CDATA[JOE TM ]]> 520 548

[0033] In another example, samples can be prepared using magnetic beads or beads used for preparing DNA samples, for example by ACROBiosystems. TM These are manufactured to bind with nucleic acid compounds. Magnetic beads are manufactured in many places, and the beads can be selected based on the characteristics of the specific sample to be analyzed. In yet another example, a sample may have both fluorescent markers / dyes and magnetic beads for further processing, utilizing optical detection systems and capacitance and magnetic arrays as further disclosed herein.

[0034] System parameters for the processing unit 206 used to drive the system including the mechanical device 202 may include start and stop times, cycle time (where cycle is the amount of time the mechanical device is in motion), acceleration of the mechanical device, speed of the system device, and many other settings such as the temperature of various heating zones, power of the capacitor zone, settings of the optical detection unit, and other parameters disclosed herein.

[0035] Now for reference Figure 3This is an illustration of an exemplary embodiment of the optical detection unit 300. The optical detection unit 300 is configured below a housing 302 containing a chip 304, in which a prepared sample is placed. Fluid channels are depicted on the chip 304, wherein mechanical devices move the sample and other fluids within the chip through interaction with pins on a rubber film on the chip surface. Figure 3 The examples do not depict heating regions or zones, nor the corresponding heating units, magnetic regions, and capacitor arrays and zones. These elements are positioned at the bottom of the chip and correspond to the various functions disclosed herein. For example, in one example, serpentine paths on the left and right sides of the chip are positioned above two separate heating elements. An element comprises a hot region or zone (95-98°C) in the fluid channel and a cold region (55-60°C) in the fluid channel. The hot region forms a first heating block or unit, and the cold region forms a second heating block or unit. This type of heating arrangement is used for the amplification process. Between the two heating regions is a voxel or optical detection region, where the optical detection unit is focused for observing fluorescent dyes within the sample.

[0036] The optical detection unit 300 is configured with an optical light-emitting element such as a diode 306, an optical detector such as a detection diode 308, and a processing unit for performing analysis on the optical detection area of ​​the chip 304. The processing unit may also be configured with a built-in timer, including a system timer for the processor, which can be used to determine peripheral device interactions. The optical light-emitting diode emits at an excitation value for a specific dye or marker within the sample to produce a result detectable by the detection diode 308. As the sample moves, fluid flows through the optical detection area, sometimes called a voxel, through a force that pushes a pin against a rubber membrane on the chip via a mechanical device, where the optical detection unit 300 performs analysis on the sample. Voxels and / or the optical detection area are the areas where the optical detection unit can perform its intended functions. This type of detection is often referred to as dynamic detection because the optical detection unit 300 performs detection while the fluid circulates, and because amplification occurs in real time. Therefore, as the fluid is circulated, the motor can be adjusted to open and close, or to change acceleration or speed. Thus, the optical detection unit serves as control of the motor, as well as for the verification and detection process of the fluid flowing between the two heating zones and the amplification status. The exemplary light-emitting diode 306 includes those made by Lumileds TM Those manufactured, for example, are LED lighting color series blue 470nm used for dyes or markers with specific wavelengths. Typical maximum current is 1 ampere, luminous flux is 35 lm, and viewing angle is 125 degrees.

[0037] The optical detection unit 300 forms an assembly that may have built-in computing power or be connected via a data cable or wirelessly to a processing unit located elsewhere on the system. Furthermore, a microcontroller and motors may be applied to the optical detection unit 300 for fine-tuning the diode angle or adjusting its separation from the chip. This improvement in the movement of the unit or assembly is known to those skilled in the art, as certain refinements in optical imaging require precise positioning of the diodes and light-emitting sources. Moreover, any number of diodes can be used; three optical light-emitting diodes are disclosed in this example. However, in other embodiments, one, two, or more may be used. Similarly, for the detection diode 308, two detection diodes are disclosed in this example; however, the number of detection diodes will vary depending on the system objectives.

[0038] Continuing, now for reference Figure 4A -E is an illustration of an exemplary embodiment of a sample moving through an optical detection region via a fluid channel, where the analog-to-digital converter (ADC) count reflects the position. Figure 4A In the diagram, sample 404 is moving through a fluid channel in the chip and is shown moving from one heated area to another by mechanical forces on the film of the chip, causing air within the chip to move the sample through the fluid channel. Sample 404 is moving toward detection area 402, where a voxel is located for better access to the optical detection unit. A light-emitting diode is positioned to direct light to the bottom side of the chip where sample 404 is moving. Since sample 404 has not yet reached optical detection area 402, the ADC count remains at baseline.

[0039] exist Figure 4B In the process, sample 404 begins to enter optical detection area 402, and the ADC count begins to increase. This increase is due to the diode reflecting away from the fluorescent dye or marker, and the diode receiving more counts. Figure 4C In this configuration, sample 404 is centered on optical detection area 402, and the ADC count reaches its peak. This reading informs the system that the sample is within the voxel, or that most of the sample is moving towards the next heated area. The motor can be controlled to run until the count drops, or pause for additional readings at optical detection area 402. Figure 4D In the process, sample 404 is moving out of the optical detection area, indicating that the sample is moving towards the heating element and away from the voxel. At this stage, the ADC count begins to drop, forming different sine waves or curves to indicate sample movement. This information is then used to notify technicians of the relative positioning of the sample to the motor, the system status, and the number of cycles executed.

[0040] exist Figure 4EIn this process, sample 404 has passed through optical detection area 402 and is entering the heating area, or within the capacitor array area, or proceeding along the fluid channel as specified. If sample 404 is not detected to have a similar curve, the system can warn technicians or pop up an error message indicating that sample 404 has not been cycled or that the operation needs to be restarted. The curve formed by the change in count over time represents the movement of the sample. ADC counting failures may include sample 404 stalling or leakage or malfunction in the chip, indicating that the system is not operating and triggering error messages or alarms via visual or audible indicators.

[0041] Now for reference Figure 5 This is an illustration of an exemplary embodiment of the chip. Fluid channels pass through the entire chip, and in this example, the orientation is unique for the capacitor array and optical detection unit. Additional chip designs and decorations can be considered, and such additional configurations form part of this disclosure, provided that the principles disclosed herein remain unchanged. The top side of the chip is a rubber membrane, which may be composed of rubber or other materials that allow bending when contacted by a motor-driven pin connected to a mechanical device (such as a mechanical roller or cam). The chip may also include several other layers, such as capacitor layers, and layers for inserting probes or other diagnostic methods.

[0042] exist Figure 5 In this example, sample initiation 512 is where the sample first enters the chip from the cartridge and processing begins. Mechanical devices move the sample through a channel sequence to a fluid hub 508. The fluid hub may contain magnets or other devices to hold the fluid within it. Next, a magnetic region 506, designed to hold portions of the sample bound to the magnetic beads within the magnetic region 506 when preparing the sample. In this system, this could be a magnetic array located under the chip for holding the sample or for other diagnostic or preparation methods.

[0043] exist Figure 5 In this example, two heating regions are disclosed: a hot region and a cold region. The temperature is defined for optimal DNA amplification. In this example, the hot region (95-98°C) and the cold region (55-60°C) are heated to specific temperatures by two heating elements. In other examples, a single heating element with variable heating zones may be used, or in additional examples, more than two heating elements may be used.

[0044] A capacitive liquid sensing array 502 is positioned in the indicated area and, in conjunction with sample preparation for magnetic beads, allows tracking of fluid within the chip's fluid channels. In this example, the capacitive array is positioned on either side of the optical detection region or the voxel, in which case the relative position of the circulation between the two heating regions can be determined. Furthermore, the capacitive liquid sensing array is positioned on the inlet and outlet sides of the heating region 504, thereby allowing complete tracking of the sample as it moves through the amplification process. The capacitive array can operate independently of the optical detection unit or in combination to detect signals or information and transmit them to a processing unit that controls motors and mechanics, as well as instruments such as a technician's screen or diagnostic tools.

[0045] Figure 6 This is an illustration of an exemplary embodiment of a capacitor array used with a chip having a capacitive sensing region. The capacitor array is positioned within a specific capacitive sensing region for detecting the presence of fluid / sample. Examples of such regions are shown in... Figure 5 The method is disclosed in the literature. Additional regions may exist, and a capacitive sensing "cladding" can be equipped to comprehensively track the sample. In this example, the sample can be prepared using magnetic beads that allow tracking using a capacitive sensing array that includes a capacitive sensor covering the capacitive sensing area on the chip. Furthermore, the capacitance of the sample fluid itself can be used to track the sample, with priority locations at the beginning and end of the heating zone and on either side of the optical detection area. Additional regions can be designated after the washing period or at other locations where occlusion or blockage may occur; this capacitive array coupled to the capacitive sensor will allow for the detection, tracking, alerting, and correction of samples and RT-qPCR operation.

[0046] Figure 7 This is a diagram illustrating an exemplary implementation of a farad detected as a sample moves across a capacitive sensing array on a capacitive sensing region of a chip through a fluid channel. In this example, a peak value of 50 nanofarads is measured and detected as the sample and / or fluid is distributed across the capacitive array, and is sensed by the capacitive array and transmitted to a processing unit. This processing unit is equipped with a processor capable of executing program instructions and defining an algorithm for motor movement based on feedback from the capacitive array and / or optical detection units.

[0047] Figure 7 The graph in the image indicates that the sample moves back and forth across the capacitive sensing array, thus the system functions as expected. In the absence of farad readings for the sample and / or fluid, the system returns to the baseline and forms a sinusoidal wave as the sample passes through. This is related to the motor speed and intensity as the sample moves from one heated zone to another. The capacitive sensing array can also provide feedback to the motor, controlling start-up and stop, as well as speed and acceleration.

[0048] Figure 8 This is a diagram illustrating an exemplary embodiment of a capacitance signal. Figure 8 In some examples, the capacitor array covers the entire serpentine channel of the heating area, such as on a chip. Figure 5 The example is defined more fully. The capacitance is shown when the sample is above the sensing array and as it gradually decreases away from the array. This allows for manipulation via noise filtering, but also allows for the use of signal strength to control motor units or provide diagnostics in RT-qPCR runs.

[0049] Figure 9 This is a graph illustrating an exemplary implementation of conventional cyclic RT-qPCR amplification time. Intrasample target amplification and cycle average timelines are disclosed.

[0050] Figure 10 This is a diagram illustrating an exemplary implementation of the functionality disclosed herein for improving the timing of RT-qPCR amplification. The system benefits from a feedback loop established by an optical detection unit and a capacitor array, coordinated with motors and mechanical devices. This allows the chip insert to remain protected while the progress of the entire cycle is monitored. Therefore, this disclosure also provides systems and methods for detecting and real-time adjusting the position of fluid volumes (e.g., samples) that move through optical detection and / or capacitive regions in the fluid channels of a chip. In one example, real-time adjustments are made through coordination with the optical detection unit and capacitor array sensors, allowing coordinated feedback and algorithms to control the motor functions of the mechanical devices. The result is reliable and rapid RT-qPCR amplification and analysis in a self-contained system that can rapidly produce results with minimal technician training.

[0051] The terms “comprises”, “comprising”, “includes”, “including”, “having”, and their morphological variations refer to “including but not limited to”.

[0052] The term "composed of" means "including and limited to".

[0053] The term "consistently made of" means that a composition, method, or structure may include additional ingredients, steps, and / or portions, provided that the additional ingredients, steps, and / or portions do not substantially alter the essential and novel characteristics of the claimed composition, method, or structure.

[0054] The term "majority" refers to two or more.

[0055] As used herein, unless the context clearly specifies otherwise, the singular forms “a,” “an,” and “the” include plural indicators. For example, the terms “a compound” or “at least one compound” can include multiple compounds, including mixtures thereof.

[0056] Throughout this application, various embodiments of the invention may be presented in a scope format. It should be understood that the scope format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Therefore, the description of the scope should be considered to have specifically disclosed all possible sub-scopes and individual numerical values ​​within that scope. For example, a description of a scope such as 1 to 6 should be considered to have specifically disclosed sub-scopes, such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and individual numbers within that scope, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the scope.

[0057] Whenever a range of numbers is indicated herein, it means to include any referenced numbers (fractions or integers) within the indicated range. The phrases “range variation / range between the first and second indicated numbers” and “range variation / range from the first indicated number to the second indicated number” are used interchangeably herein and mean to include the first and second indicated numbers and all decimals and integers in between.

[0058] It should be understood that, for clarity, certain features of the invention described in the context of individual embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the invention described in the context of a single embodiment may also be provided individually, or in any suitable sub-combination, or appropriately provided in any other described embodiment of the invention. Certain features described in the context of various embodiments should not be considered essential features of those embodiments unless the embodiment is invalid without these elements.

[0059] Although the invention has been described in conjunction with specific embodiments thereof, it will be apparent to those skilled in the art that many alternatives, modifications, and variations will be readily apparent. Therefore, it is intended to include all such alternatives, modifications, and variations falling within the spirit and broad scope of the appended claims.

[0060] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference in their entirety, as if each individual publication, patent, or patent application were expressly and individually indicated to be incorporated herein by reference. Furthermore, any reference or identification of any reference in this application should not be construed as an admission that such reference is available as prior art to the invention. The use of section headings should not be construed as necessarily limiting.

Claims

1. A system for detecting and real-time adjusting the position control of a fluid volume moving through an optical detection region in a fluid channel, the system comprising: Mechanical devices; Consumable box, A chip, disposed within the consumable box, wherein the chip and the consumable box are consumables, allowing for single-use and disposal in accordance with various regulations and / or protocols, the chip comprising; (i) A fluid channel, wherein the mechanical device controls the movement of the fluid volume based on the analysis results of the processing unit; and (ii) Optical detection area; One or more independent heating blocks, wherein the chip is at least partially positioned on the one or more independent heating blocks such that the fluid channel is adjacent to the one or more independent heating blocks; An optical detection unit is located below the consumable box containing the chip, and the optical detection unit includes: (i) Optical light-emitting elements; (ii) Optical detectors; and (iii) A processing unit for analyzing the fluid position in the optical detection area of ​​the chip.

2. The system of claim 1, wherein the mechanical device is a top roller, and the top roller is positioned to press a pin onto the rubber membrane of the chip for fluid control.

3. The system according to claim 1 further includes a housing for accommodating the mechanical device, the chip, and the optical detection unit.

4. The system according to claim 1, wherein the chip further comprises voxels within the optical detection area.

5. The system of claim 1, further comprising a motor configured to respond to the processing unit, wherein the motor controls the mechanical device, which in turn applies fluid control through the fluid channel of the chip.

6. The system of claim 5, wherein the processing unit further includes a timer for recording the time and position of the motor.

7. The system of claim 1, wherein the optical light-emitting element transmits light to the detection area of ​​the chip, wherein an optical detector is provided for detecting fluorescent dye of fluid in the detection area from the fluid channel of the chip.

8. The system of claim 1 further includes a hot region on the fluid channel and a cold region on the fluid channel, wherein the hot region is on one of the one or more independent heating blocks, and the cold region is on different heating blocks of the one or more independent heating blocks.

9. The system of claim 1, wherein the processing unit is on a single PCB board.

10. The system according to claim 1 further comprises three optical light-emitting diodes and two dual-band filters.

11. The system of claim 1, further comprising a capacitive sensing region and a capacitive sensing array on the chip, the capacitive sensing array being configured to detect the presence of a sample and to communicate electrically with the processing unit.

12. A method for detecting and real-time adjusting the position control of a fluid volume moving through an optical detection region in a fluid channel, the method comprising: Samples were prepared using fluorescent markers; A chip is configured within a consumable cartridge, the chip and the consumable cartridge being consumables that allow for single-use and disposal in accordance with various regulations and / or protocols to receive the sample, wherein the chip has a fluid channel for the flow of the sample and an optical detection area that allows optical light to be transmitted to the fluid channel; The sample is applied to the chip; Activate the optical detection unit located below the consumable box containing the chip, wherein activating the optical detection unit is configured to illuminate one or more LEDs and activate the optical detection diodes; The chip is applied to two heating arrays; A mechanical device is activated to press down on a region of the chip, wherein the pressed region causes fluid within the chip, including the sample, to move along the fluid channels and through the two heating arrays; The presence of the sample and the signal output of the sample within the optical detection area are detected by the optical detection unit. as well as The motion control script is adjusted based at least on the signal output, wherein the motion control script adjusts at least the start and / or stop of the mechanical device.

13. The method of claim 12, wherein the chip is configured to receive a sample, and the chip is further configured to have: (i) a rubber film on the surface of the chip, (ii) a plurality of fluid channels, (iii) a hot fluid channel region, (iv) a cold fluid channel region and (v) one or more capacitive sensing regions.

14. The method of claim 12, further comprising applying the chip to a capacitive liquid sensing array, wherein the capacitive liquid sensing array measures changes in capacitance to detect the presence of fluid in a specific region of the fluid channel on the chip.

15. The method of claim 12, wherein adjusting the motion control script further adjusts at least the speed and / or acceleration of the mechanical device.

16. A method for detecting and real-time adjusting the position control of a fluid volume moving into a fluid channel, the method comprising: Samples were prepared using magnetic beads that adsorbed nucleic acid compounds; A chip is configured within a consumable cartridge, the chip and the consumable cartridge being consumables that allow for single-use and disposal in accordance with various regulations and / or protocols to receive the sample, wherein the chip has a fluid channel for the flow of the sample and the fluid channel passes through two separate heating arrays; The sample is applied to the chip; An array of capacitors is applied along a segment of the fluid channel of the chip; A mechanical device is activated to press down on a region of the chip, wherein the pressed region causes fluid within the chip, including the sample, to move along the fluid channels and through the two separate heating arrays; as well as The capacitance change in the fluid channel at the capacitor array is detected by a capacitance sensor; It also includes adjusting a motion control script based at least on signal output from the capacitor array, wherein the motion control script adjusts the speed and / or acceleration of the mechanical device pressing the pin into the region of the chip.

17. The method of claim 16, wherein a capacitance change is detected by the capacitance sensor, indicating that the sample exists in the fluid channel in a specific region of the capacitor array.

18. The method of claim 16, further comprising preparing the sample with a fluorescent label.

19. The method of claim 18, further comprising activating an optical detection unit, wherein activating the optical detection unit is configured to illuminate one or more LEDs and activate an optical detection diode.

20. The method of claim 16, further comprising activating a hot region and a cold region on the fluid channel, wherein the hot region is a first heating block and the cold region is a second heating block.

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