Cell sorting chip, device, and method based on deterministic lateral displacement by dielectrophoresis

Abstract

We provide a cell sorting chip, device, and method based on deterministic lateral displacement by dielectrophoresis. The chip includes a microchannel (4) with one inlet (4.1), a direct channel, and two outlets. The bottom of the direct channel is equipped with three electrode array pairs: a focusing electrode group (1), a sorting electrode group (2), and a separation electrode group (3). The focusing electrode group is used for cell focusing and signal detection, the sorting electrode group is used for single-cell sorting, and the separation electrode group is used for single-cell separation. This chip can achieve high-speed, accurate, and complete sorting of target cells.

Description

[Technical field]

[0001] The present invention is in the field of biotechnology and instrumental science, and specifically relates to cell sorting chips, devices and methods based on deterministic lateral displacement by dielectrophoresis. [Background technology]

[0002] Traditionally, in biology, biological cells are mainly characterized at the population cell level, but the average measurement of a heterogeneous population relies on cell culture, which cannot embody the differences of individual cells, and the majority of cells in nature are still difficult to culture. Functional identification and directional sorting of single cells does not depend on cell proliferation, so it can directly skip the cell culture step and can also reflect the heterogeneity of cells. Fluorescence-activated cell sorting (FACS) is a high-throughput single-cell analysis technique, and the application of this technology has significantly improved the identification efficiency of single cells. However, since the majority of cells themselves have weak or no fluorescence effect, this method generally requires the addition of fluorescent labels. In addition, when identifying functional components in cell colonies, important cell phenotypes are often only roughly recognized or completely unknown (i.e., ``unknown'' cell phenotypes), and there are no other bio-labels, and the purpose of these studies is often precisely to find these unknown but important phenotypes (and their bio-labels). Therefore, the barrier of ``detecting unknown cell phenotypes'' currently significantly limits the application of cell sorting techniques such as FACS to single-cell research. Single-cell Raman spectroscopy (SCRS) is a highly efficient technology for identifying intracellular chemical information, and can provide information on the composition and structure of intracellular compound molecules. Raman spectroscopy does not require any labeling when obtaining the chemical fingerprint spectrum of the entire single cell, and can identify changes in cell type, physiological characteristics and phenotype of living cells, and can track and select "unknown cell phenotypes" by utilizing changes in cellular Raman signals. The realization of Raman activated cell sorting (RACS) based on SCRS provides a new idea for solving the above-mentioned barriers.

[0003] In the prior art, there are a series of techniques for Raman activated cell sorting, such as optical clamping, laser jet coupling, etc., which belong to the static version. Although these systems are simple and practical, their throughput is too low, which hinders the application of Raman spectrum technology to high-throughput sorting. In order to improve the throughput of single cell sorting, Raman activated droplet sorting systems and sorting methods that can achieve high-throughput sorting have also appeared, but this kind of technical solution uses water-in-oil droplets, which have complex flow paths and are difficult to handle, and still cannot fully meet the needs of practical use. Summary of the Invention [Problem to be solved by the invention]

[0004] In view of the above problems, the present invention proposes a cell sorting chip, device, and method based on deterministic lateral displacement by dielectrophoresis, which realizes the sorting of single cells by applying a non-uniform electric field using a simple chip structure. [Means for solving the problem]

[0005] In order to achieve the above-mentioned object, one aspect of the present invention provides a cell sorting microfluidic chip based on deterministic lateral displacement by dielectrophoresis, which includes a microchannel including one inlet, a direct passage and two outlets, and three sets of electrode array pairs, including a focusing electrode group, a sorting electrode group and a separation electrode group, are integrated at the bottom of the direct passage, the focusing electrode group is used for cell focusing and signal detection, the sorting electrode group is used for single-cell sorting, and the separation electrode group is used for single-cell separation.

[0006] The focusing electrodes and the separating electrodes include at least One electrode pair The selection electrode group includes: Electrode Pair or it may be one electrode.

[0007] The focusing electrode group includes: Electrode pair Each of the arrays includes at least one Electrode pairThe electrodes in the array are parallel to each other, and one end of the electrodes on a single electrode array is connected, Electrode pair The electrodes in the array are distributed at intervals and have an included angle.

[0008] The electrode has two sides that resemble a ">" in shape.

[0009] The included angle between the electrodes is a+b, and the electrode angles a and b are in the range of 0 to 90 degrees, and preferably 15 to 45 degrees.

[0010] a is the angle between one side of the electrode and the center line, and b is the angle between the other side of the electrode and the center line.

[0011] In another preferred embodiment, the angles a and b are not the same.

[0012] In another preferred embodiment, the angles a and b are the same.

[0013] The separating electrode group and the focusing electrode group have the same structure, but the included angle directions are different, and the included angle between the separating electrode and the center line is a.

[0014] The electrode shape of the separated electrodes resembles "<".

[0015] The width L of the electrode pair can be flexibly adjusted according to the width of the microchannel.

[0016] The electrode width D and the electrode pitch d can be adjusted according to the size of the cell / particle, but both the electrode width D and the electrode pitch d are larger than the maximum dimension of the cell / particle.

[0017] In another preferred embodiment, the center lines of the electrode pairs of the focusing electrode group and the separating electrode group are not on the same straight line.

[0018] In another preferred example, the distance c between the center lines of two electrode pairs is determined by the electrode width D and the electrode pitch d.

[0019] In another preferred example, the width L of the electrode pair is 1 mm, the electrode width D is 25 microns, the inter-electrode distance d is 25 microns, the electrode angles a and b are both 30 degrees, and the distance c between the center lines of the two electrode pairs is 35 microns.

[0020] In another preferred example, the material of the electrode group is ITO, carbon, graphene, or a metal electrode.

[0021] In another preferred example, the height of the electrode is from 10 nanometers to 10 microns, preferably from 50 to 150 nanometers.

[0022] In another preferred example, the material of the carrier that integrates the electrode pairs on the bottom of the chip is silicate glass, quartz glass, calcium fluoride glass, polydimethylsiloxane (PDMS) or polymethylmethacrylate (PMMA), and is preferably quartz glass.

[0023] In another preferred example, the electrode array group is externally connected to a function generator for applying a high-frequency AC voltage to the electrodes.

[0024] In another preferred embodiment, the response time of the selection electrodes is 50 milliseconds.

[0025] By periodically applying a high-frequency AC voltage to the electrodes, the particles in the fluid are made to flow along the electrodes. By applying a non-uniform electric field, cells / particles in the solution that are difficult to polarize become more easily polarized, and the cells are subjected to a dielectrophoretic force perpendicular to and oriented toward the electrodes, and a fluid propulsion force parallel to and oriented in the flow direction, and the cells move along the electrodes due to the combined force of the two.

[0026] The width of the DC path does not have a theoretical upper limit, and the channel width can reach a desired focusing effect by increasing the electrode array.

[0027] In another preferred embodiment, the width of the direct current path is 1 to 2 millimeters.

[0028] Another aspect of the present invention provides a cell sorting device based on deterministic lateral displacement by dielectrophoresis, comprising: A microfluidic chip; A function generator for generating sinusoidal AC voltages of different frequencies and voltages; a computer for controlling the execution of the program and the acquisition and collection of signals by the associated optical instruments, for controlling the output voltage, frequency, duty cycle, and other parameters of the function generator, and for controlling the switching of the relay; a relay for controlling switching of the control circuit; and a fluid drive device as a driving force source for driving the fluid into the microchannel.

[0029] In another preferred embodiment, the fluid drive includes a syringe pump or a gravity drive.

[0030] Another aspect of the present invention provides a method for cell sorting based on deterministic lateral displacement by dielectrophoresis, comprising the steps of: 1) injecting cells by using a fluid drive to direct a fluid containing cells through an inlet of the microfluidic chip; 2) connecting the focusing electrodes to the output terminal of the function generator via a conductor and periodically applying a high-frequency voltage to the focusing electrodes to focus the cells in the fluid at the position of the signal monitoring point of the electrode angle; 3) detecting an optical signal by aligning the optical signal collection point with the monitoring point 5, and processing the detected signal with a computer program to detect a signal that is determined to be a target cell or not; 4) a step of selecting cells in which, if the cells are target cells, a relay is triggered to communicate with the selection electrodes and output a high frequency voltage, thereby shifting the target cells from their centerline position along the selection electrodes, and, if the cells are determined by the program to be non-target cells, the relay is not turned on by the program, thereby allowing the non-target cells to continue moving along the original centerline position of the focusing electrodes without being shifted; 5) Separating non-target cells and target cells in the fluid by periodically applying a high frequency voltage to the separation electrodes, thereby causing the non-target cells and target cells in the fluid to flow toward different outlets.

[0031] In another preferred embodiment, the cell is one or more of a biological cell such as a yeast cell, an E. coli cell, or a HeLa cell.

[0032] In another preferred embodiment, the flow rate of the fluid is 0.01 to 50 microliters / minute, the magnitude of the flow rate being related to the width and height of the direct passage, and preferably, the injection flow rate is 10 to 40 microliters / minute.

[0033] Taking yeast as an example, the yeast is washed three times with pure water, and the pure water is used as the injection buffer solution, and the optimum parameters are a sinusoidal AC voltage of 16 volts and a frequency of 10 megahertz.

[0034] In another preferred embodiment, the duration of the applied high-frequency voltage is adjusted based on parameters such as the fluid flow rate in the flow field, and the loading time is adjusted based on the distance c or more between the target cell and the center line of the two pairs of electrodes. to make It is necessary that the following is satisfied.

[0035] The detection of an optical signal in step 3) is detection of a Raman signal or detection of a fluorescent signal.

[0036] There is a delay time between the detection of the optical signal in step 3) and the sorting in step 4), and the delay time is calculated and determined by the formula: Interval time = Liquid flow length / Liquid flow velocity. The liquid flow length refers to the actual distance traveled by the detected liquid from the signal detection point to the sorting operation point.

[0037] If the sorting and separation electrodes are not triggered, the system method can be used as a flow cytometry detection, but a signal is detected for each cell flowing through the detection point. Effect of the Invention

[0038] The present invention has the following beneficial effects: 1) The chip microchannel has a simple structure, a single direct channel, and is easy to handle. 2) The response time of the sorting electrodes is on the order of milliseconds, allowing for high-throughput sorting efficiency. 3) By integrating the focusing electrodes, the efficiency of cell capture and detection can be guaranteed, and the cell injection flow rate can be significantly improved, effectively preventing cells from settling when injected into the microtube, improving the injection efficiency, and effectively extending the time that the system operates stably. [Brief description of the drawings]

[0039] [Figure 1] Schematic diagram of a cell sorting chip based on deterministic lateral displacement by dielectrophoresis. Reference symbols: 1 focusing electrode group, 2 sorting electrode group, 3 separation electrode group, 4 microchannel, 4.1 inlet, 4.2 collection port, 4.3 waste port, 5 signal detection point. [Diagram 2] Schematic diagram of each parameter of the electrodes. a and b each represent the electrode angle, d represents the distance between the electrodes, D represents the electrode width, c represents the distance between the center lines of two electrode pairs, and L represents the width of the electrode pair. [Diagram 3] FIG. 13 is a structural schematic diagram of another electrode group. [Figure 4]Schematic diagram of a cell sorting device based on deterministic lateral displacement by dielectrophoresis. Reference symbols: 1 microfluidic chip, 2 function generator, 3 computer, 4 relay, 5 injection pump. [Diagram 5] FIG. 1 is a schematic diagram showing the process of cell sorting based on deterministic lateral displacement by dielectrophoresis. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] Detailed Description of the Invention The present invention will now be described in more detail with reference to the drawings and examples. It should be understood that these examples are merely illustrative of the present invention and are not intended to limit the scope of the present invention.

[0041] The structure of the microfluidic chip is shown in Figure 1. The microfluidic chip includes a microchannel 4, which includes an inlet 4.1, a direct passage, a collection port 4.2, and a waste port 4.3. Three pairs of electrode arrays, including a focusing electrode group 1, a sorting electrode group 2, and a separation electrode group 3, are integrated at the bottom of the microchannel 4. The focusing electrode group is used for focusing cells and detecting signals, the sorting electrode group is used for sorting single cells, and the separation electrode group is used for separating single cells.

[0042] Figure 2 is a schematic diagram of each parameter of the electrodes. a and b both represent the electrode angle, d represents the distance between the electrodes, D represents the electrode width, c represents the distance between the center lines of the two electrode pairs, and L represents the width of the electrode pairs.

[0043] Figure 3 is a schematic diagram of the structure of another electrode group. Note that the selection electrode group has only one electrode. EXAMPLES

[0044] Example 1 Selection of high oil-producing yeast cells by Raman flow cytometry

[0045] Experimental preparation: One milliliter of cultured yeast cells was placed in a centrifuge tube and centrifuged at 5,000 rpm for 5 minutes, the supernatant was discarded, 1 milliliter of pure water was added and resuspended, then centrifuged again and centrifuged at 5,000 rpm for 5 minutes. After washing three times, 1 milliliter of pure water was added and resuspended. Depending on the concentration of the bacteria, this was diluted 1,000 to 10,000 times, a certain concentration of surfactant (e.g., PF127 at a final concentration of 0.5%) was added to prevent the cells from adsorbing to the electrode, and after uniform mixing, it was injected into a syringe.

[0046] The Raman measuring instrument was turned on, and the laser light was turned on so that the laser light coincided with the signal monitoring point 5 .

[0047] The structure of the microfluidic chip used is as shown in Figure 3. The parameters of the electrodes in the microfluidic chip were: electrode pair width L 1 mm, electrode width D 25 microns, interelectrode distance d 25 microns, electrode angles a and b both 30 degrees, and center line distance c of the two electrode pairs 35 microns.

[0048] A microfluidic chip 1, a function generator 2, a computer 3, a relay 4, and an injection pump 5 were connected as shown in Fig. 4, and a program organized in the computer was executed to make output channel 2 of the function generator control output channel 1, to make channel 2 output a square wave with a voltage of 20 volts and a frequency of 5 Hz, and to make output channel 1 output a sine wave with a voltage of 16 volts and a frequency of 10 MHz. At the same time, the collection and analysis of the Raman signal were controlled by the program, and when a target cell was detected, the on-state of relay 4 was controlled by the program.

[0049] During the test: The above-mentioned treated bacterial liquid was injected into the microchannel using the injection pump 5, and the yeast cells in the liquid were gradually moved to the tip of the electrode pair under the focusing action of the focusing electrodes. The flow rate of the microchannel was about 40 microliters / min, the electrode current parameters were voltage 16 volts, frequency 10 megahertz, and the focusing trap efficiency reached more than 95%.

[0050] When the yeast cells moved to the signal monitoring point, the Raman spectrum was collected and the collected spectrum was simply analyzed by the program. If it was a target cell, the Raman spectrum reflected the characteristic peak of fats and oils, so the program determined that it was a target cell, and the program actively controlled the relay to turn on the sorting electrodes and move the target cell down by presetting the delay and load duration. If it was a non-target cell, the Raman spectrum did not show the characteristic peak of fats and oils, so the relay was not activated, there was no voltage load on the sorting electrodes, and the cell continued to move along the center line.

[0051] The role of the separation electrodes is to displace the cells on either side of the centerline of the separation electrodes further from the centerline, so that target cells are positioned below the channel and non-target cells are positioned above the channel.

[0052] Finally, the high oil producing yeast cells were collected at the collection port, and the low or no oil producing yeast cells were in the waste port.

[0053] Test End: The microfluidic chip channels were rinsed twice with absolute ethanol and then placed in an oven to dry and be reused.

[0054] Example 2 Detection of high-oil-producing yeast cells by Raman flow cytometry

[0055] Experimental preparation: One milliliter of cultured yeast cells was placed in a centrifuge tube and centrifuged at 5,000 rpm for 5 minutes, the supernatant was discarded, 1 milliliter of pure water was added and resuspended, then centrifuged again and centrifuged at 5,000 rpm for 5 minutes. After washing three times, 1 milliliter of pure water was added and resuspended. Depending on the concentration of the bacteria, this was diluted 1,000 to 10,000 times, a certain concentration of surfactant (e.g., PF127 at a final concentration of 0.5%) was added to prevent the cells from adsorbing to the electrode, and after uniform mixing, it was injected into a syringe.

[0056] The Raman measuring instrument was turned on, and the laser light was turned on so that the laser light coincided with the signal monitoring point 5 .

[0057] The structure of the microfluidic chip used is as shown in Figure 3. The parameters of the electrodes in the microfluidic chip were: electrode pair width L 1 mm, electrode width D 25 microns, interelectrode distance d 25 microns, electrode angles a and b both 30 degrees, and center line distance c of the two electrode pairs 35 microns.

[0058] A microfluidic chip 1, a function generator 2, a computer 3, a relay 4, and an injection pump 5 were connected as shown in Figure 4, and a program organized in the computer was executed to make the output channel 2 of the function generator control the output channel 1, to make the channel 2 output a square wave with a voltage of 20 volts and a frequency of 5 Hz, and to make the output channel 1 output a sine wave with a voltage of 16 volts and a frequency of 10 MHz. At the same time, the collection and analysis of the Raman signal were controlled by the program.

[0059] The above-mentioned treated bacterial liquid was injected into the microchannel using the injection pump 5, and the yeast cells in the liquid were gradually moved to the tip of the electrode pair under the focusing action of the focusing electrodes. The flow rate of the microchannel was about 40 microliters / min, the electrode current parameters were voltage 16 volts, frequency 10 megahertz, and the focusing trap efficiency reached more than 95%.

[0060] When the yeast cells migrated to the signal monitoring point, Raman spectra were collected and the collected spectra were simply analyzed by the program.

[0061] Finally, the yeast cells after detection were collected at a collection port.

[0062] Test End: The microfluidic chip channels were rinsed twice with absolute ethanol and then placed in an oven to dry and be reused. The collected Raman spectra could then be later analysed to obtain relevant information, such as the proportion of high oil-producing yeast cells.

[0063] Example 3 Flow cytometric sorting of fluorescent cells

[0064] Experimental preparation: One milliliter of the cultured E. coli containing a partially expressible fluorescent protein was placed in a centrifuge tube and centrifuged at 5,000 rpm for 5 minutes, the supernatant was discarded, 1 milliliter of pure water was added and resuspended, then centrifuged again and centrifuged at 5,000 rpm for 5 minutes. After washing three times, 1 milliliter of pure water was added and resuspended. Depending on the concentration of the bacteria, this was diluted 1,000 to 10,000 times, a certain concentration of surfactant (e.g., PF127 at a final concentration of 0.5%) was added to prevent the cells from adsorbing to the electrode, and after uniform mixing, it was injected into a syringe.

[0065] The fluorescent imager was turned on and the laser light was turned on so that it coincided with the signal monitoring point 5 .

[0066] The structure of the microfluidic chip used is as shown in Figure 3. The width L of the electrode pair was 1 mm, the electrode width D was 25 microns, the interelectrode distance d was 25 microns, the electrode angles a and b were both 15 degrees, and the distance c of the center lines of the two electrode pairs was 50 microns.

[0067] A microfluidic chip 1, a function generator 2, a computer 3, a relay 4, and an injection pump 5 were connected as shown in Fig. 4, and a program organized in the computer was executed to make the output channel 2 of the function generator control the output channel 1, to make channel 2 output a square wave with a voltage of 20 volts and a frequency of 0.2 Hz, and to make output channel 1 output a sine wave with a voltage of 16 volts and a frequency of 1 MHz. At the same time, the collection and analysis of the fluorescent signal were controlled by the program, and when a target cell was detected, the on-state of relay 4 was controlled by the program.

[0068] During the test: The above-mentioned treated bacterial liquid was injected into the microchannel using the injection pump 5, and the E. coli cells in the liquid were gradually moved to the tip of the electrode pair under the focusing action of the focusing electrodes. The flow rate of the microchannel was about 10 microliters / min, the electrode current parameters were voltage 16 volts, frequency 1 megahertz, and the focusing trap efficiency reached over 90%.

[0069] When the E. coli cells move to the signal monitoring point, the E. coli containing the fluorescent protein are excited by the laser light and emit a fluorescent signal, and this fluorescent signal is collected by the computer, and the relay is actively controlled by the program to turn on the selection electrodes, and the target cells are moved down by presetting the delay and load duration. Since the E. coli not containing the fluorescent protein do not excite a fluorescent signal when passing the monitoring point, the program is controlled not to activate the relay, there is no voltage load on the selection electrodes, and the cells continue to move along the center line.

[0070] The role of the separation electrodes is to displace the cells on either side of the centerline of the separation electrodes further from the centerline, so that target cells are positioned below the channel and non-target cells are positioned above the channel.

[0071] Finally, the fluorescent E. coli were collected at the collection port, and the non-fluorescent E. coli cells were in the waste port.

[0072] Test End: The microfluidic chip channels were rinsed twice with absolute ethanol and then placed in an oven to dry and be reused.

[0073] In light of the above teachings of the present invention, it will be understood that those skilled in the art can make various changes or modifications to the present invention, and the equivalents thereof are within the scope defined by the claims appended hereto.

Claims

1. A microchannel including one inlet, a direct passage, and two outlets; At the bottom of the direct passage, three sets of electrode array pairs, namely, a focusing electrode group (1), a selection electrode group (2), and a separation electrode group (3), are sequentially stacked; The focusing electrode group (1) is used for focusing cells and detecting signals, The sorting electrode group (2) is used for sorting a single cell, The separation electrode group (3) is used for separating single cells. A cell sorting microfluidic chip based on deterministic lateral displacement by dielectrophoresis, comprising: The focusing electrode group (1) and the separation electrode group (3) each include at least one electrode pair, and the selection electrode group (2) is at least one electrode; Each electrode of the focusing electrode group (1) has two sides in the shape of ">", Each electrode of the separated electrode group (3) is in the shape of "<", A microfluidic chip characterized in that the center lines of the electrode pairs of the focusing electrode group (1) and the separation electrode group (3) are not collinear, so that individual cells from the sorting electrode group (2) can be further displaced from the center line of the electrode pair of the separation electrode group (3) within the separation electrode group (3).

2. the included angle between the two sides of each electrode of the focusing electrode group (1) is expressed as a+b, which is the sum of electrode angle a and electrode angle b, where electrode angle a is the angle between one side of the electrode and the center line of the electrode pair of the focusing electrode group (1), electrode angle b is the angle between the other side of the electrode and the center line of the electrode pair of the focusing electrode group (1), and the ranges of a and b are each 0 to 90 degrees; The microfluidic chip described in claim 1, characterized in that the included angle between the two sides of each electrode of the separated electrode group (3) is expressed as a+b, which is the sum of electrode angle a and electrode angle b, where electrode angle a is the angle between one side of the electrode and the center line of the electrode pair of the separated electrode group (3), electrode angle b is the angle between the other side of the electrode and the center line of the electrode pair of the separated electrode group (3), and the ranges of electrode angles a and b are each 0 to 90 degrees.

3. 3. The microfluidic chip according to claim 2, wherein the electrode angles a and b each range from 15 to 45 degrees.

4. The microfluidic chip according to claim 3 , wherein the electrode angle a and the electrode angle b are not the same value.

5. The microfluidic chip according to claim 3 , wherein the electrode angle a and the electrode angle b have the same value.

6. The microfluidic chip according to any one of claims 2 to 5, wherein the separation electrode group (3) and the focusing electrode group (1) have the same structure, the directions of the included angles are different, and the value of the included angle between the center lines of the electrode pairs of the selection electrode group (2) and the focusing electrode group (1) is the same as the electrode angle α.

7. The microfluidic chip according to claim 1 , wherein the material of the electrode group is ITO, carbon, graphene, or a metal electrode.

8. The microfluidic chip according to any one of claims 1 to 7, a function generator for generating sinusoidal AC voltages of different frequencies and voltages; a computer for controlling the execution of the program and the acquisition and collection of signals by the associated optical instruments, for controlling the parameters output by the function generator, and for controlling the switching of the relay; a relay for controlling switching of the control circuit; a fluid drive unit as a driving force source for driving the fluid to enter the microchannel; A cell sorting device based on deterministic lateral displacement by dielectrophoresis, comprising:

9. The cell sorting apparatus according to claim 8 , wherein the fluid drive device comprises a syringe pump or a gravity drive device.

10. Utilizing the microfluidic chip according to any one of claims 1 to 7 or the device according to claim 8 or 9, 1) injecting cells by using a fluid drive to direct a fluid containing cells through an inlet of the microfluidic chip; 2) connecting the focusing electrodes to the output terminal of the function generator via a conductor and periodically applying a high-frequency voltage to the focusing electrodes to focus the cells in the fluid at the position of the signal monitoring point of the electrode angle; 3) detecting an optical signal by aligning the optical signal collection point with the monitoring point, and processing the detected signal with a computer program to detect a signal that is determined to be a target cell or not; 4) if the cell is a target cell, a relay is triggered to communicate with the selection electrode group and a high frequency voltage is outputted, thereby shifting the target cell along the selection electrode group from the center line of the electrode pair of the focusing electrode group, and if the cell is determined by the program to be a non-target cell, the relay is not turned on by the program, thereby selecting the non-target cell, so that the non-target cell continues to move along the original center line of the electrode pair of the focusing electrode group without being shifted; 5) applying a high-frequency voltage periodically to the separation electrode group to move non-target cells and target cells in the fluid away from the center line of the electrode pair of the separation electrode group, thereby separating the cells to flow toward different outlets; A method for sorting cells based on deterministic lateral displacement by dielectrophoresis, comprising:

11. 11. The method for cell sorting based on deterministic lateral displacement by dielectrophoresis according to claim 10, wherein the cells are one or more of yeast cells, E. coli cells, or HeLa cells.

12. 12. The method for cell sorting based on deterministic lateral displacement by dielectrophoresis according to claim 10, wherein the flow rate of the fluid is 0.01 to 50 microliters per minute.

13. 12. The method for sorting cells based on deterministic lateral displacement by dielectrophoresis according to claim 10 or 11, wherein the detection of an optical signal in step 3) is detection of a Raman signal or detection of a fluorescent signal.

Images