A microfluidic chip, a preparation method, an analysis device and an analysis method

Abstract

The application belongs to the technical field of treating microorganisms or particles with electromagnetic sound energy, and particularly relates to a microfluidic chip, a preparation method, an analysis device and an analysis method. The microfluidic chip comprises a main channel, two or more sample inlets which are in communication with the main channel, a focusing electrode and a deflection electrode which are arranged at the bottom of the main channel, the focusing electrode is used for gathering particles, and the deflection electrode is used for directional movement of the particles. The microfluidic chip of the application is provided with multiple sample inlets, so that multiple fluids can enter the main channel at the same time. By adjusting the flow rates of the fluids in different sample inlets, the fluid in the main channel can controllably enter a sample outlet or a waste outlet, directional movement of the particles is realized, in the movement process, a specific detection point is selected, specific detection is completed, on-line cleaning based on dielectricity is realized, compatibility of multiple complex samples in the upstream is realized, and the downstream is conveniently connected to more in-depth analysis.

Description

[0001] This invention is an application that invokes priority, the priority basis of which includes: application number 202210101266.X, patent title "Chip, Manufacturing Method, Analytical Device Method and Single Cell Multiphenotypic Analysis Application", application date January 27, 2022. Technical Field

[0002] This application belongs to the technical field of treating microorganisms or particles with energy such as electromagnetic sound, and in particular relates to a microfluidic chip, preparation method, analytical device and analytical method. Background Technology

[0003] Cells are the most basic structural and functional building blocks of most organisms, and their study has garnered significant attention. In cell biology, cell populations are classified and studied based on characteristics such as their tissue of origin, morphology, and secretions. This research is crucial for understanding human drug metabolism, organismal behavior, and signal transduction. However, these methods are all based on cell populations—large numbers of cells—and studies in cell morphology and metabolomics represent the average level of these large cell groups. These methods cannot be used to study cellular differences. In recent years, scientists have increasingly emphasized single-cell research, and its role has become increasingly important. It not only relates to drug metabolism and new drug development but also provides valuable insights into cellular molecular mechanisms and pathways. Therefore, comprehensive analysis of individual cells is a vital technique in modern biological and medical research.

[0004] Currently, various technologies have been developed for cell analysis and detection.

[0005] Single-cell Raman spectroscopy (SCRS) is one such technique. It offers advantages such as high resolution, label-free operation, non-destructive nature, molecular specificity, and culture-free operation. SCRS is a highly efficient technique for identifying intracellular chemical substances, providing information on the molecular composition and structure of compounds within cells. Raman spectroscopy obtains the chemical fingerprint of the entire single cell without requiring any labels; chemical bonds are detectable. Therefore, it can identify cell types, physiological characteristics, and phenotypic changes in living cells, and can use changes in cell Raman signals to track and sort "unknown cell phenotypes." However, the acquisition of single-cell Raman spectra is affected by the Raman background of liquids. The presence of amino acids or sugars in the environment of the cell will affect the measured values. Therefore, certain pretreatment of the cells is necessary before detecting the cells or bacteria.

[0006] There are two main traditional cell pretreatment methods. One method involves centrifuging the cells or bacteria, followed by multiple precipitation steps, removal of the supernatant, and washing with buffer. The other method is based on filtration, where cells or bacteria are filtered onto filter paper, then the cells or bacteria on the filter paper are transferred, collected, and washed. Raman spectroscopy is then performed after these pretreatment steps.

[0007] However, processing complex samples is more cumbersome; and traditional offline cleaning methods can affect cell activity and state, making it difficult to meet the needs of online detection, such as monitoring the state of bacteria during bacterial fermentation; at the same time, inactivated cells cannot be used for downstream analysis. Summary of the Invention

[0008] To achieve compatible analysis of various complex samples from upstream sources and facilitate downstream cell analysis, this application provides the following technical solutions for a microfluidic chip, its fabrication method, and an online analysis device.

[0009] The term "microparticle" as used in this invention refers to particles that can be suspended in a non-organic phase solution (such as an aqueous phase) and pass through the microfluidic chip of this invention. These include particles of biological origin and particles of non-biological origin, such as eukaryotic cells, prokaryotic cells, single-celled organisms, virus particles, organelles, particles formed by biological macromolecules, drug particles, drug carrier particles, liposomes, polymer particles, etc.

[0010] Firstly, this application provides a microfluidic chip, which is implemented using the following technical solution.

[0011] A microfluidic chip includes a main channel, with two or more sample inlets connected to the main channel. A focusing electrode and a deflection electrode are provided at the bottom of the main channel. The focusing electrode is used to aggregate microparticles, and the deflection electrode is used to directionally move microparticles.

[0012] By adopting the above technical solution and setting up multiple injection channels, multiple samples can enter the main channel simultaneously. By adjusting the flow rate of the fluid in different injection channels, the fluid in the main channel can be controlled to enter the outlet channel or waste channel, thereby achieving the directional movement of particles.

[0013] Preferably, the focusing electrode is a pointed interdigitated electrode, and the pointed end of the pointed interdigitated electrode points away from the injection channel.

[0014] By adopting the above technical solution, the pointed end of the focusing electrode points in the same direction as the fluid flow, which can successfully capture particles in the fluid and make the particles in the fluid accurately gather at the pointed end, making it convenient to set up spectrometer detection equipment at the pointed end.

[0015] Preferably, there are two injection channels, namely a first injection channel and a second injection channel.

[0016] By adopting the above technical solution, a fluid actuator is used to inject the fluid containing particles and the buffer solution into the main channel respectively. Only the substances entering through these two channels are in the main channel. By controlling their flow rate, the fluid, except for particles, will not flow into the outlet channel, thus realizing the directional movement of cells and / or particles in the fluid.

[0017] Secondly, this application provides a method for fabricating a microfluidic chip, which is implemented using the following technical solution.

[0018] Fabricating the microfluidic chip includes the following steps:

[0019] (1) Fix the focusing electrode and the deflection electrode on the substrate; cut out the main channel and the injection channel on the double-sided tape;

[0020] (2) Adhere one side of the double-sided adhesive to the base layer;

[0021] (3) The top cover layer is attached to the other side of the double-sided adhesive.

[0022] By adopting the above technical solution, the conductive electrodes enable the particles in the main channel to be subjected to dielectric force. Etching the main channel, sample inlet channel, sample outlet channel, and waste sample channel onto the double-sided adhesive facilitates the setting of the length, width, and height parameters of these channels and also simplifies processing. Thirdly, this application provides an analytical device, implemented using the following technical solution.

[0023] An analytical apparatus, a particle carrying and capturing device, for carrying and capturing particles;

[0024] A fluid actuator, connected to the particle carrier and capture device, is used to drive fluid into the particle carrier and capture device;

[0025] A spectrometer, wherein the spectral information acquisition end of the spectrometer is aligned with the particle capture point of the particle-carrying capture device, for detecting spectral information at the particle capture point;

[0026] A genome analysis device, connected to a particle carrier and capture device, is used to detect particle information;

[0027] A computer, connected to the spectrometer and the genome analysis device, is used to control the spectrometer and the genome analysis device, and to record and analyze information obtained from the spectrometer and the genome analysis device.

[0028] By adopting the above technical solution, a feasible device is provided for realizing coupled analysis of spectral and genomic detection.

[0029] Preferably, the microfluidic chip includes a sample outlet channel; a fluid driver for driving fluid into the sample outlet channel; a function generator for providing AC power to the microfluidic chip; a spectrometer with its spectral information acquisition end aligned with the sharp end of the focusing electrode; a genome analysis device connected to the sample outlet channel; and a computer for controlling the function generator, the spectrometer, and the genome analysis device, and for collecting information.

[0030] By employing the above technical solution, a voltage is applied to the focusing and deflection electrodes of the microfluidic chip via a function generator, causing the microparticles within the microfluidic chip to be subjected to dielectric forces. The entire analytical apparatus is then connected to a computer via a spectrometer and other subsequent analytical devices, enabling the system to be turned on, controlled, and to acquire, store, and analyze data.

[0031] Preferably, the sample outlet is connected to one end of the post-processing tube, and the other end of the post-processing tube is connected to the oil pool. The post-processing tube is connected to a first feeding tube and a second feeding tube, and the droplets in the oil pool are used for genome analysis.

[0032] By adopting the above technical solution, subsequent secondary analysis can be achieved to complete genome analysis, thereby realizing the coupling of spectral detection and genome analysis.

[0033] Preferably, the particle carrying and capturing device is a particle screening chip;

[0034] The particle screening chip includes an oil storage tank, a sample storage tank, a main channel, and a second sample inlet channel. The two ends of the main channel are connected to the oil storage tank and the second sample inlet channel, respectively. The sample storage tank is a sealed hollow three-dimensional structure, and the outlet of the sample storage tank is connected to the main channel through a branch channel.

[0035] By adopting the above technical solution, the purpose of this application is to achieve the purpose of particle movement and positioning by any means that can move particles, including various means of particle movement such as force, sound, electricity, heat, light, and magnetism, so as to perform spectral detection.

[0036] Fourthly, this application provides an analysis method, which is implemented using the following technical solution.

[0037] An analytical method using the aforementioned microfluidic chip and analytical device includes the following steps:

[0038] (1) Connecting device:

[0039] Connect the function generator to the focusing electrode and the deflection electrode respectively;

[0040] The spectrometer's spectral information acquisition end is aligned with the main channel;

[0041] The sample outlet of the microfluidic chip is connected to the genome analysis device;

[0042] The computer is connected to the function generator, spectrometer, and genome analysis equipment, respectively.

[0043] (2) Sample injection: The buffer solution is introduced into the main channel through the first injection channel; the fluid containing particles is introduced into the main channel through the second injection channel;

[0044] (3) Focusing: Applying a voltage to the focusing electrode to cause particles in the fluid to aggregate;

[0045] (4) Optical detection: Turn on the spectrometer and transmit the detected signal to the computer program for analysis and processing;

[0046] (5) Deflection: Applying a voltage to the deflection electrode to deflect the particles in the fluid;

[0047] (6) Secondary detection: Add enzyme and barcoad-containing gel bead suspension to the liquid flowing out of the sample channel, mix to form droplets, and perform single-cell sequencing on the droplets to achieve genome analysis.

[0048] By employing the above technical solution, microparticles are aggregated under the action of a focusing electrode, spectral detection points are set, and then the microparticles flow out through the sample outlet under the action of a deflection electrode. The cells flowing out of the sample outlet are processed and then subjected to genome detection. Single-cell multiphenotype analysis is completed. Preferably, a microparticle screening chip is used, and the spectral signal acquisition end of the spectrometer is set at the junction of the branch channel and the main channel; the computer connects the spectrometer and the genome analysis device, and a fluid driver is used to inject the sample into the sample storage cell; buffer solution is injected into the first inlet.

[0049] By adopting the above technical solution, it is possible to couple the spectral and genomic detection of the same single cell.

[0050] In summary, this application includes at least one of the following beneficial technical effects:

[0051] 1. The microfluidic chip of this application sets up multiple sample inlet channels, allowing multiple fluids to enter the main channel simultaneously. By adjusting the flow rate of the fluid in different sample inlet channels, the fluid in the main channel can be controllably sent to the sample outlet channel or the waste sample channel, realizing the directional movement of particles. During the movement, a specific detection point is selected to complete a specific detection.

[0052] 2. The analytical device of this application applies voltage to the focusing electrode and deflection electrode of the microfluidic chip through a function generator, causing the microparticles within the microfluidic chip to be subjected to dielectric force. Other devices for subsequent analysis, such as spectrometers and genomic analysis equipment, are connected to a computer to enable the operation, control, data acquisition, storage, and analysis of the entire analytical device.

[0053] 3. This application is based on dielectric online cleaning, which enables compatibility with a variety of complex samples from upstream and facilitates deeper analysis downstream. Attached Figure Description

[0054] Figure 1 This is a schematic diagram of a microfluidic chip structure.

[0055] Figure 2 This is a schematic diagram showing the parameter characteristics of the focusing electrode and the deflection electrode.

[0056] Figure 3 This is a schematic diagram of the microchannel structure in Example 2.

[0057] Figure 4 This is a diagram of the microfluidic chip layer structure.

[0058] Figure 5 This is a schematic diagram of single-cell sequencing coupled with spectral signal detection.

[0059] Figure labeling: 10, Main channel; 11, Focusing electrode; 12, Deflection electrode; 13, First inlet channel; 14, Second inlet channel; 15, Outlet channel; 16, Waste channel; 17, Microparticle; 18, Spectral information acquisition end; 22, Post-processing tube; 23, First feed tube; 24, Oil pool; 25, Second feed tube; 26, Barcoad-containing gel bead; 27, Top cap layer; 28, Double-sided adhesive; 29, Base layer; 30, Third inlet channel; 31, Second outlet channel; 35, Droplet; a: Upper electrode angle; b: Lower electrode angle; d: Distance between electrodes; D: Electrode width; L: Width of electrode pair. Detailed Implementation

[0060] The following is in conjunction with the appendix Figure 1-5 This application will be described in further detail below.

[0061] Example 1

[0062] This embodiment is a microfluidic chip. (Refer to...) Figure 1 The microfluidic chip has a main channel 10, one end of which is connected to two injection channels. The injection channels described in this application can be straight, zigzag, curved, or irregular in shape. In this embodiment, the two injection channels are the first injection channel 13 and the second injection channel 14, with shapes as follows: Figure 1 As shown, the widths of the different inlet channels in this application can be different. In this embodiment, the widths of the first inlet channel 13 and the second inlet channel 14 are the same. An outlet channel 15 and a waste channel 16 are connected at the other end of the main channel 10. The first inlet channel 13 and the outlet channel 15 are on the same side of the main channel 10, while the second inlet channel 14 and the waste channel 16 are on the other side of the main channel 10. A focusing electrode 11 and a deflection electrode 12 are provided at the bottom of the main channel 10.

[0063] The focusing electrode 11 and the deflecting electrode 12 are arranged sequentially along the direction from the inlet channel to the outlet channel 15. The focusing electrode 11 is a pointed interdigitated electrode, which is bent and has an included angle, and its shape is similar to ">". The pointed interdigitated electrodes are arranged in pairs. This application has at least one pair of focusing electrodes 11. The two electrodes in each pair are parallel to each other and spaced apart. One electrode in each electrode pair extends from one side of the self-channel 10 to the bottom of the main channel 10, and the other electrode in the electrode pair extends from the other side of the self-channel 10 to the bottom of the main channel 10. If multiple pairs of focusing electrodes 11 are provided, the electrodes located on the same side of the main channel 10 are connected at one end. In this embodiment, there are two electrode pairs serving as focusing electrodes 11, and the pointed ends of the pointed interdigitated electrodes point away from the inlet channel.

[0064] The deflection electrodes 12 also appear in pairs. Each electrode is a straight plate-shaped electrode, inclined relative to the main channel 10, and its shape is similar to " / ". The two electrodes of the paired deflection electrodes 12 are parallel to each other and spaced apart, and the shape of the parallel and spaced deflection electrode pair 12 is similar to " / / ". In the embodiment, there is one electrode pair serving as the deflection electrode 12.

[0065] See attached document Figure 2 In the figure, a represents the upper electrode angle; b represents the lower electrode angle; d represents the distance between electrodes; D represents the electrode width; and L represents the width of the electrode pair. In this application, a and b both range from 0-90°. The number of focusing electrodes 11 is at least one pair, and the number of deflecting electrodes 12 is at least one, both of which achieve the purpose of this application. In this embodiment, there are two pairs of focusing electrodes 11 and two deflecting electrodes 12. The electrode pair width L is 1 mm, the electrode width D is 25 micrometers, the distance d between electrodes is 25 micrometers, the upper electrode angle a is 30 degrees, and the lower electrode angle b is also 30 degrees.

[0066] In this application, the electrode heights of the focusing electrode 11 and the deflection electrode 12 are 10 nanometers to 10 micrometers, with 50-150 nanometers being more preferred. In this embodiment, the heights of both the focusing electrode 11 and the deflection electrode 12 are 100 nanometers.

[0067] The length of the main channel 10 in this application is not limited. The longer the channel, the more focusing electrodes 11 can be added, resulting in a better focusing effect. In this embodiment, the length of the main channel 10 is 1.5 mm.

[0068] The implementation principle of this embodiment is as follows: Cell suspensions contain substances such as glucose and sucrose. When performing single-cell detection and analysis, these substances can interfere with and affect the detection results. Therefore, single cells need to be separated from the cell suspension during detection and analysis. This application achieves online cleaning of the microparticles 17 by setting two injection channels and introducing buffer pure water into the first injection channel 13. A focusing electrode 11 and a deflection electrode 12 are set at the lower part of the main channel, and a high-frequency alternating current is periodically applied to the focusing electrode 11 and the deflection electrode 12, causing the microparticles 17 in the fluid to move along the electrodes. This is because in a solution that is not easily polarized, the microparticles 17 are easily polarized by the application of a non-uniform electric field, and will experience a dielectric force perpendicular to and pointing towards the electrodes. At the same time, the microparticles 17 will also experience a fluid driving force parallel to and pointing towards the flow direction. The resultant force of these two forces will move the microparticles 17 along the electrodes.

[0069] Example 2

[0070] This embodiment is a microfluidic chip. (Refer to...) Figure 3 This embodiment is basically the same as Embodiment 1, except that the microfluidic chip includes three inlet channels (first inlet channel 13, second inlet channel 14, and third inlet channel 30), two outlet channels (first outlet channel 15 and second outlet channel 31), and one waste channel 16. The width of each inlet channel can be set differently; in this embodiment, the first inlet channel 13, second inlet channel 14, and third inlet channel 30 have the same width. Fluid is introduced into the three inlet channels at different flow rates, and the outflow channel is controlled by the flow rate to be either the first outlet channel 15, the second outlet channel 31, or the waste channel 16. In this embodiment, the second inlet channel 14 is used to introduce cell suspension, the third inlet channel 30 is used to introduce buffer solution (pure water), the first inlet channel 13 is used to introduce lysis solution to achieve online cleaning and lysis of microparticles 17, the first outlet channel 15 is connected to the detection device, the lysis solution, water and culture medium flow out through the second outlet channel 31, and the residual substances of cell suspension are discharged through the waste channel 16.

[0071] In this embodiment, a fluid actuator is used to inject the fluid containing particles and the buffer solution into the main channel respectively. Only these two types of substances enter the main channel. By controlling their flow rate, the fluid, except for the particles, will not flow into the sample outlet channel 15, thus achieving the directional movement of the particles in the fluid.

[0072] The implementation principle of this embodiment is basically the same as that of embodiment 1. Different cells are sorted and sent to different outlet channels, either the first outlet channel 15 or the second outlet channel 31. The sorted particles 17 can be analyzed through different outlet channels.

[0073] Example 3

[0074] This embodiment describes a method for fabricating a microfluidic chip. (Refer to...) Figure 4 The microfluidic chip consists of three parts: a base layer 29 with electrode structure, a double-sided adhesive layer 28, and a top cover layer 27.

[0075] Fabrication of substrate 29: ITO electrodes with the desired electrode structure are etched onto the ITO glass. In this embodiment, the etched electrodes are... Figure 4 The shape shown Figure 4 The heights of the focusing electrode 11 and deflection electrode 12 are shown in the schematic diagram; their actual height is 100 nanometers. (The ratio of electrode height to glass height in the diagram does not represent the actual height ratio.) The substrate glass is marked with the names of the sample inlet channel, first sample inlet channel 13, second sample inlet channel 14, sample outlet channel 15, and sample outlet channel 16. The glass of the substrate layer 29 is quartz glass.

[0076] Fabrication of 28 layers of double-sided tape: The required channel structure is cut out on the double-sided tape 28 using a tool. The channel structure in this embodiment is as follows: Figure 4 The channel structure in it.

[0077] Double-sided adhesive 28 is applied to the base layer 29 with electrode structure, aligned with the electrode structure. Then, a top cover layer 27 is placed on the double-sided adhesive 28 layer. The double-sided adhesive 28 bonds the upper and lower glass layers to form the main channel 10 and other channel structures.

[0078] The implementation principle of the microfluidic chip fabrication method in this embodiment is as follows: Two glass plates are bonded together using double-sided adhesive 28, which provides high transparency. The main channel 10, the first sample inlet channel 13, the second sample inlet channel 14, the sample outlet channel 15, and the waste sample channel 16 are etched onto the double-sided adhesive 28, forming channels with the upper and lower glass plates through which microparticles 17 can pass. Electrodes are etched onto the substrate glass layer 29, offering strong controllability in electrode shape, high operability, and firm adhesion.

[0079] Example 4

[0080] This embodiment describes the specific operational steps of the Raman sequencing analysis method for colon cancer cell microparticle 17.

[0081] The cultured colon cancer cell microparticles 17 were digested, washed, and resuspended in a mixed solution containing glucose and sucrose. This solution can maintain the osmotic activity of the cell microparticles 17 and simultaneously achieve dielectric capture of the cell microparticles 17 under a dielectric field.

[0082] Reference Figure 1 and Figure 5Turn on the Raman instrument and the laser, aligning the laser with the spectral information acquisition end 18. After a period of warm-up, calibrate the system. Connect the various devices: pump the cell suspension containing cell microparticles 17 from the second inlet channel 14 into the main channel 10 of the microfluidic chip via a fluid driver (injection pump); use pure water as a buffer and pump it into the main channel 10 of the microfluidic chip from the first inlet channel 13. Adjust the flow rate ratio of pure water in the first inlet channel 13 to the fluid in the second inlet channel 14, making the flow rate of pure water in the first inlet channel 13 20 μL / min and the flow rate of fluid in the second inlet channel 14 10 μL / min. At this time, glucose in the cell suspension will not flow into the outlet channel 15. Use a function generator to apply a dielectric field to the focusing electrode 11 and the deflection electrode 12 (the focusing electrode 11 and the deflection electrode 12 can be voltaged simultaneously or separately): the voltage is 16 volts and the frequency is 10 MHz. Raman signal detection begins. After detection, the Raman spectrum of particle 17 is saved for analysis. The flow rate of the first injection channel 13 is twice that of the second injection channel 14, ensuring that the buffer flowing into the second injection channel 14 does not affect the detection of cells in the fluid of the first injection channel 13. Waste channel 16 is collected as waste liquid. The outlet of the outlet channel 15 is connected to the post-processing tube 22. The enzyme added through the second feeding tube 25 and the barcoad-containing gel bead suspension added through the first feeding tube 23 are mixed and form droplets 35 at the oil pool 24. The resulting droplets are then subjected to a series of sequencing procedures for particle 17, including reverse transcription, cDNA amplification, library construction, and high-throughput sequencing. This enables genomic analysis of the measured particle 17. The data from the saved Raman spectrum and the data obtained from the genomic sequencing are compared to perform multiphenotypic analysis of particle 17. This allows for simultaneous online coupled detection of spectral and genomic analyses of the same single cell, achieving multiphenotypic analysis of the spectral and genomic data of particle 17.

[0083] The implementation principle of the analytical method in this embodiment is as follows: Cell suspensions contain substances such as glucose and sucrose. When performing single-cell detection and analysis, these substances can interfere with and affect the detection results. Therefore, it is necessary to separate the single cells from the cell suspension during detection and analysis. This application achieves online cleaning of cell particles 17 by setting two sample inlet channels. A focusing electrode 11 and a deflection electrode 12 are arranged at the lower part of the main channel, and a high-frequency alternating current is periodically applied to the focusing electrode 11 and the deflection electrode 12, causing the particles 17 in the fluid to move along the electrodes. This is because in a solution that is not easily polarized, when a non-uniform electric field is applied, the cells / particles are easily polarized and will experience a dielectric force perpendicular to and pointing towards the electrodes. At the same time, the cell particles 17 will also experience a fluid driving force parallel to and pointing towards the flow direction. The resultant force of these two forces will move the cell particles 17 along the electrodes.

[0084] The signal acquisition area of ​​the spectrometer is aligned with the sharp end of the focusing electrode 11. The sharp end passes through the transparent base layer 29 and top cover layer 27, enabling the acquisition and analysis of spectral information of the particles 17 in the main channel 10 without damaging the structure of the particles 17, which facilitates the subsequent processing of the particles 17 and secondary analysis.

[0085] The microparticles 17 emerging from the sample outlet 15 were processed into droplets, and a series of microparticle 17 sequencing processes were performed, including reverse transcription, cDNA amplification, library construction, and high-throughput sequencing.

[0086] The above are all preferred embodiments of this application, and are not intended to limit the scope of protection of this application. Therefore, all equivalent changes made in accordance with the structure, principles and principles of this application should be covered within the scope of protection of this application.

Claims

1. A microfluidic chip, comprising a main channel (10), characterized in that: Two or more sample inlets are connected to the main channel (10). The bottom of the main channel (10) is provided with a focusing electrode (11) and a deflection electrode (12). The focusing electrode (11) is used to aggregate particles. The deflection electrode (12) is used for the directional movement of particles; When two sample inlets are provided in connection with the main channel (10): Two injection channels, namely the first injection channel (13) and the second injection channel (14), are connected to the outlet channel (15) and the waste channel (16) at the other end of the main channel (10); the first injection channel (13) and the outlet channel (15) are on the same side of the main channel (10), and the second injection channel (14) and the waste channel (16) are on the other side of the main channel (10); By setting up two injection channels, buffer solution is introduced into the first injection channel (13) to achieve online cleaning of particles (17); When three sample inlets are provided in connection with the main channel (10): The three inlet channels are the first inlet channel (13), the second inlet channel (14), and the third inlet channel (30). At the other end of the main channel (10), there are two outlet channels and one waste channel (16). The two outlet channels are the first outlet channel (15) and the second outlet channel (31). Fluids are introduced into the three inlet channels at different flow rates. The outflow channel of each fluid is controlled by the flow rate to be the first outlet channel (15), the second outlet channel (31), or the waste channel (16). The cell suspension is introduced into the second inlet channel (14), the buffer solution is introduced into the third inlet channel (30), and the lysis solution is introduced into the first inlet channel (13) to achieve online cleaning and lysis of the microparticles (17). The first outlet channel (15) is connected to the detection equipment. The lysis solution, water and culture medium flow out through the second outlet channel (31), and the residual substances of the cell suspension are discharged through the waste channel (16). The focusing electrode (11) is a pointed interdigitated electrode, and the pointed end of the pointed interdigitated electrode points away from the injection channel.

2. A method for fabricating a microfluidic chip, comprising fabricating the microfluidic chip as described in claim 1, characterized in that, Includes the following steps: (1) The focusing electrode (11) and the deflection electrode (12) are fixed on the substrate layer (29); the main channel (10) and the sample inlet channel are cut out on the double-sided adhesive tape (28); (2) Adhere one side of the double-sided adhesive (28) to the base layer (29); (3) The top cover layer (27) is attached to the other side of the double-sided adhesive (28).

3. An analytical apparatus, characterized in that, include: A particle carrying and capturing device for carrying and capturing particles; The particle carrying and capturing device is the microfluidic chip as described in claim 1; A fluid actuator, connected to the particle carrier and capture device, is used to drive fluid into the particle carrier and capture device; A spectrometer, wherein the spectral information acquisition end of the spectrometer is aligned with the particle capture point of the particle-carrying capture device, for detecting spectral information at the particle capture point; A genome analysis device, connected to a particle carrier and capture device, is used to detect particle information; A computer, connected to the spectrometer, or to both the spectrometer and the genome analysis device, is used to control the spectrometer and the genome analysis device and to record and analyze information obtained from the spectrometer and the genome analysis device.

4. The analytical apparatus as described in claim 3, characterized in that: The microfluidic chip is powered by an AC power source provided by a function generator. The spectral information acquisition end (18) of the spectrometer is aligned with the sharp end of the focusing electrode (11); The genome analysis device is connected to the sample outlet (15).

5. The analytical apparatus according to claim 4, characterized in that: The sample outlet (15) is connected to one end of the post-processing tube (22), and the other end of the post-processing tube (22) is connected to the oil pool (24). The post-processing tube (22) is connected to a first feeding tube (23) and a second feeding tube (25). The droplets in the oil pool (24) are used for genome analysis.

6. An analytical method, characterized in that: Using the microfluidic chip according to claim 1 includes the following steps: (1) Connecting device: Connect the function generator to the focusing electrode (11) and the deflection electrode (12) respectively; The spectrometer's spectral information acquisition end (18) is aligned with the main channel (10); The computer is connected to both the function generator and the spectrometer. (2) Sample injection: The buffer solution is introduced into the main channel (10) through the first injection channel (13); the fluid containing particles is introduced into the main channel (10) through the second injection channel (14); (3) Focusing: Applying a voltage to the focusing electrode (11) causes the particles in the fluid to aggregate; (4) Optical detection: Turn on the spectrometer and transmit the detected signal to the computer program for analysis and processing; (5) Deflection: Apply voltage to the deflection electrode (12) to deflect the particles in the fluid; (6) Secondary detection: Add the liquid flowing out of the sample outlet (15) to the enzyme and the barcoad-containing gel bead (26) suspension, mix them to form droplets, and perform single-cell sequencing on the droplets to achieve genome analysis.

Images