A method for sorting fluorescent coded microspheres

Abstract

The present invention discloses a method for sorting fluorescent coded microspheres, wherein different coded microspheres are mixed in a sample to be tested, and corresponding different target molecules are captured respectively to obtain a liquid to be tested; the liquid to be tested is controlled by a fluid control system to sequentially enter the gathering area, real-time detection area, light pattern conversion area, and sorting and separation area of ​​a photoelectric chip to achieve the sorting of fluorescent coded microspheres and corresponding molecules to be tested. The present invention uses a light-conducting material in combination with a light pattern design to change the direction of the dielectrophoretic force by changing the electric field distribution on the plane, thereby changing the force on the coded microspheres, achieving the purpose of positioning, capturing, releasing, and moving the coded microspheres, and improving the efficiency of coded microsphere sorting and immunoassay.

Description

Technical Field

[0001] The invention belongs to the field of biological detection, and in particular relates to a method for sorting fluorescent coded microspheres. Background Art

[0002] The detection of many cancers is closely related to the concentration of cytokines, and sometimes one cancer cell will cause the release of multiple cytokines. In clinical practice, the detection of the number and concentration of interleukins in the patient's serum can be used as an indicator for early cancer diagnosis and after treatment. For example, the measurement of cytokines such as IL-1, IL-2, IL-6, IL-8, IL-10, IL-15, IL-17, etc. can be used for clinical detection of liver cancer, lung cancer, pancreatic cancer and ovarian cancer. For lung cancer, breast cancer and ovarian cancer, most treatment times are less than 1 year from the discovery of the disease to ineffective treatment. If multiple cytokines can be detected in serum, the accuracy of cancer diagnosis and the survival rate of cancer patients can be improved as early as possible.

[0003] Single-cell capture and analysis can accurately obtain specific single cells from a large number of cells, and realize a variety of uses such as disease diagnosis and hybridoma cell screening. Microfluidics technology (also known as microfluidics) can manipulate and process cells in the medium at the single-cell scale, and realize a series of processes such as cell capture, culture, analysis, processing, and screening.

[0004] Encoded microspheres are common carriers for single cell capture. In addition, encoded microspheres can also capture molecules. Encoded microspheres can be combined with target analytes to achieve encoding of target analytes, which is convenient for subsequent detection and sorting. There are many ways to encode microspheres, such as encoding microspheres based on color and intensity.

[0005] The positioning of particles is also important for particle detection and sorting. Generally speaking, the dielectrophoretic force is often used to locate small particles at the micrometer scale, which means that particles will induce charges on the surface under the action of an uneven electric field, and the particles will move to the place where the electrode strength is large when they are subjected to a positive dielectrophoretic force, and move to the place where the electrode strength is small when they are subjected to a negative dielectrophoretic force. By adjusting the frequency applied by the outside world, the direction of the force on the particles can be adjusted.

[0006] The general metal electrophoretic force design uses a micro-electromechanical process to first deposit a metal layer on the glass surface, and then use photoresist to define the electrode shape. After etching and development, a metal pattern (conductive layer) can be formed at the micron scale, and then electrophoretic force is generated in the micron channel (microfluidic pipeline). However, the pattern of the metal electrode is fixed, the shape of the electric field is also fixed, the flexibility is insufficient, and the process is complex, requiring special manufacturing technology and training. Summary of the invention

[0007] The purpose of the present invention is to provide a method for sorting fluorescent coded microspheres, to achieve the positioning, grabbing, releasing, moving and detecting of the coded microspheres, to provide a better method for sorting fluorescent coded microspheres, thereby improving the efficiency of immunoassay.

[0008] The present invention discloses a method for sorting fluorescent coded microspheres, comprising the following steps:

[0009] Mix different coded microspheres in the sample to be tested, and capture the corresponding different target molecules to obtain the test solution;

[0010] The fluid to be tested is controlled to enter the optoelectronic chip through a fluid control system. The optoelectronic chip includes a first transparent electrode and a second transparent electrode. A fluid channel is formed between the first transparent electrode and the second transparent electrode. A photoconductive layer is provided on a side of the second transparent electrode close to the fluid channel. The photoconductive layer can change conductivity under light. The optoelectronic chip is connected to an electrical signal control system, and the aggregation and positioning of fluorescent coded microspheres can be achieved in the light pattern irradiation area.

[0011] By projecting a light pattern onto the optoelectronic chip, the optoelectronic chip is divided into a gathering area, a real-time detection area, a light pattern conversion area, and a sorting and separation area in sequence; the gathering area arranges the scattered fluorescent coded microspheres into the real-time detection area; in the real-time detection area, the fluorescent coded microspheres are arranged linearly, and the fluorescent coded microspheres are irradiated with excitation light and the fluorescence is collected to identify the fluorescent coded microspheres; in the light pattern conversion area, a corresponding light pattern channel is generated according to the fluorescent coded microsphere information obtained in the real-time detection area, and the fluorescent coded microspheres are guided to the corresponding sorting and separation area; the sorting and separation area includes a plurality of flow channels to separate and guide the fluorescent coded microspheres;

[0012] The liquid to be tested passes through the aggregation area, the real-time detection area, the light pattern conversion area, and the sorting and separation area in sequence to achieve the sorting of the fluorescent coded microspheres and the corresponding molecules to be tested.

[0013] In some embodiments, the distance between the first transparent electrode and the second transparent electrode is 50-200 micrometers.

[0014] In some embodiments, the first transparent electrode and the second transparent electrode are ITO glass.

[0015] In some embodiments, the light-guiding layer is made of TiOPc or hydrogenated amorphous silicon.

[0016] In some embodiments, the fluorescent encoded microspheres are made of two fluorescent dyes in different mixing ratios.

[0017] In some embodiments, the fluorescently encoded microspheres contain iron-containing nanoparticles that can be controlled by an external magnetic field.

[0018] In some embodiments, the surface of the fluorescently encoded microspheres is modified to specifically capture different molecules.

[0019] In some embodiments, the focusing area includes two strip-shaped light patterns, and both of the two strip-shaped light patterns lead to the real-time detection area.

[0020] Compared with the traditional technology of generating electrophoretic force, the present invention combines the light pattern with the photoconductive material to make the generated light pattern become an electrode with variable position and generate dielectrophoretic force, which can produce good grasping for the microspheres in the liquid. It is not only relatively simple in operation, but also simplifies the experimental process and improves the efficiency of particle grasping. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 A schematic diagram of the device used in the present invention;

[0022] Figure 2 The structure diagram of the optoelectronic chip design of the present invention;

[0023] Figure 3 This is an example diagram of the light pattern array in the present invention;

[0024] Figure 4 This is an example diagram of the light pattern array in the present invention;

[0025] Figure 5 This is a schematic diagram of the first part of the light pattern of Example 2 of the present invention;

[0026] Figure 6 It is a schematic diagram of the second part of the light pattern of Example 2 of the present invention;

[0027] Figure 7 This is a schematic diagram of the third part of the light pattern of Embodiment 2 of the present invention;

[0028] Figure 8 Schematic diagram of the overall light pattern of Example 2 of the present invention.

[0029] Figure numerals: 1-optical system, 2-fluid control system, 3-electric signal control system, 4-photoelectric chip, 5-fluid collecting device, 6-light pattern emission source, 7-first excitation light, 8-second excitation light, 9-first glass substrate, 10-first transparent electrode, 11-photoconductive layer, 12-second transparent electrode, 13-second glass substrate, 14-signal generator, 15-light pattern. DETAILED DESCRIPTION

[0030] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0031] It should be noted that, unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of the technical field of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0032] The present invention provides a fluorescent microsphere detection device, the detection device comprising:

[0033] An optoelectronic chip is provided with a first transparent electrode and a second transparent electrode, a fluid channel is formed between the first transparent electrode and the second transparent electrode, a photoconductive layer is provided on a side of the second transparent electrode close to the fluid channel, and the photoconductive layer can change conductivity under light;

[0034] A fluid control system for controlling the injection of the liquid to be tested into the optoelectronic chip;

[0035] An optical system capable of sending a light pattern signal and an excitation light signal to the optoelectronic chip, wherein the light pattern signal is used to adjust the conductivity of the photoconductive layer in the irradiated area, and the excitation light is used to excite the coded microspheres in the liquid to be tested to generate a fluorescent signal;

[0036] An electrical signal control system for adjusting electrical signals in the optoelectronic chip;

[0037] An image processing system for collecting and processing the fluorescent signal emitted by the optoelectronic chip.

[0038] An array of light patterns is formed on a large area of ​​photoconductive material. Due to the characteristics of the photoconductive material, the resistance of the photoconductive material decreases where light is irradiated, and electrons move from the conductive layer below to the upper surface of the photoconductive layer, resulting in conductive and non-conductive areas on the plane. The target particles are fixed on the plane using the dielectrophoresis effect of microparticles at the microfluidic scale to record the fluorescence of the particles themselves and the fluorescence of molecules captured on the particle surface. The fluorescence signals generated by the coded microspheres include the fluorescence of the fluorescent coded microspheres themselves, as well as the fluorescence signals of the fluorescent molecules modified or captured on the surface of the coded microspheres.

[0039] The detection device of the coded microspheres of the present invention can realize accurate positioning of the coded microspheres and improve the detection efficiency. The light-conducting material used in the present invention can change the direction of the dielectrophoretic force by changing the pattern of the light pattern and the electric field distribution on the plane, thereby changing the force on the coded microspheres, achieving the purpose of positioning, grabbing, releasing, and moving the coded microspheres, and improving the efficiency of immunoassay.

[0040] The present invention also proposes a method for sorting fluorescent coded microspheres, using the detection device for the coded microspheres as described above, comprising the following steps:

[0041] Different coded microspheres are mixed into the sample to capture the corresponding different target molecules;

[0042] The working area of ​​the optoelectronic chip is divided into a gathering area, a real-time detection area, a light pattern conversion area, and a sorting and separation area through light pattern projection;

[0043] The gathering area includes two crossed bars, the intersection of which is located in the real-time detection area, and the scattered fluorescent coded microspheres are arranged into a straight line and gathered in the real-time detection area;

[0044] In the real-time detection area, the fluorescent coded microspheres are arranged linearly, and the fluorescent coded microspheres are irradiated with excitation light and fluorescence is collected to identify the fluorescent coded microspheres;

[0045] In the light pattern conversion area, a corresponding light pattern channel is generated according to the fluorescence coded microsphere information obtained in the real-time detection area, and the fluorescence coded microsphere is directed to the corresponding sorting and separation area;

[0046] The sorting and separation zone includes a plurality of flow channels for separating and exporting the introduced fluorescent coded microspheres.

[0047] In some embodiments, the present invention discloses a method for sorting fluorescently encoded microspheres, comprising the following steps:

[0048] Mix different coded microspheres in the sample to be tested, and capture the corresponding different target molecules to obtain the test solution;

[0049] The fluid to be tested is controlled to enter the optoelectronic chip through a fluid control system. The optoelectronic chip includes a first transparent electrode and a second transparent electrode. A fluid channel is formed between the first transparent electrode and the second transparent electrode. A photoconductive layer is provided on a side of the second transparent electrode close to the fluid channel. The photoconductive layer can change conductivity under light. The optoelectronic chip is connected to an electrical signal control system, and the aggregation and positioning of fluorescent coded microspheres can be achieved in the light pattern irradiation area.

[0050] By projecting a light pattern onto the optoelectronic chip, the optoelectronic chip is divided into a gathering area, a real-time detection area, a light pattern conversion area, and a sorting and separation area in sequence; the gathering area arranges the scattered fluorescent coded microspheres into the real-time detection area; in the real-time detection area, the fluorescent coded microspheres are arranged linearly, and the fluorescent coded microspheres are irradiated with excitation light and the fluorescence is collected to identify the fluorescent coded microspheres; in the light pattern conversion area, a corresponding light pattern channel is generated according to the fluorescent coded microsphere information obtained in the real-time detection area, and the fluorescent coded microspheres are guided to the corresponding sorting and separation area; the sorting and separation area includes a plurality of flow channels to separate and guide the fluorescent coded microspheres;

[0051] The liquid to be tested passes through the aggregation area, the real-time detection area, the light pattern conversion area, and the sorting and separation area in sequence to achieve the sorting of the fluorescent coded microspheres and the corresponding molecules to be tested.

[0052] Compared with the traditional technology of generating dielectrophoretic force, the present invention combines the light pattern with the photoconductive material to make the generated light pattern become an electrode with variable position and generate dielectrophoretic force, which can produce good grasping for the microspheres in the liquid. It is not only relatively simple in operation, but also simplifies the experimental process and improves the efficiency of particle grasping.

[0053] Compared with the method of using metal electrodes to generate electric fields, the production method is simpler and the material cost is lower. The photoconductive layer material can be TiOPc, and it only needs to be spin-coated to form a photoconductive layer with a thickness of less than 1 micron. Compared with the traditional use of metal electrodes, it is necessary to enter a clean room, deposit a conductive layer, and repeatedly expose and develop to establish a metal pattern. The spin coating method is simpler and costs less, and does not require expensive machinery and equipment.

[0054] Compared with flow cytometer, flow cytometer passes the particles to be tested through the laser irradiation area at an extremely fast speed, measures the fluorescence of the particles, and distinguishes the number, size, and brightness of the particles. Due to its high speed, errors are prone to occur in measurement. Although it is known for its high throughput, its accuracy still needs to be improved.

[0055] Using the patented method, the moving particles can be quickly fixed at the position to be measured, presented in an array, and observed under a microscope after laser irradiation, and then the fluorescence is recorded, which further improves the accuracy. Secondly, the originally scattered particles can be gathered into a straight line after the light pattern, and then identified and sorted, so as to effectively screen the captured particles.

[0056] In addition, the most important consumable in this device is the microfluidic chip, but it has a low production cost and a simple structure. It can be used to locate and grasp various particles. It is easy to operate and can effectively reduce costs.

[0057] The device used in the fluorescent coding microsphere sorting method of the present invention is as follows Figure 1 As shown, it includes an optical system 1, a fluid control system 2, an electrical signal control system 3, an optoelectronic chip 4, a fluid collection device 5 and an image processing system (not shown).

[0058] like Figure 2 As shown, the optoelectronic chip 4 is provided with a first glass substrate 9, a first transparent electrode 10 attached to the first glass substrate 9, a second glass substrate 13, a second transparent electrode 12 attached to the second glass substrate 13, and a photoconductive layer 11 attached to the second transparent electrode 12. A fluid channel is formed between the first transparent electrode and the photoconductive layer, which is connected to the fluid control system 2. The photoconductive layer 11 is located on the side of the second transparent electrode 12 close to the fluid channel, and can change the conductivity under light. The optoelectronic chip of the present invention has the characteristics of microfluidics.

[0059] The distance between the first transparent electrode 10 and the second transparent electrode 12 is between 50 and 200 micrometers, preferably about 100 micrometers. The first transparent electrode 10 and the second transparent electrode 12 are respectively connected to the signal generator 14 in the electric signal control system 3 .

[0060] The transparent electrode is preferably an ITO electrode, but may also be other materials such as nanosilver. The photoconductive layer will conduct electricity after being illuminated, and will be an electric field barrier when not illuminated. After being illuminated, the conductivity of the photoconductive layer in the illuminated portion increases, and the charge is transferred from the ITO glass to the photoconductive layer, and the illuminated area is like a virtual electrode. In some embodiments, the material of the photoconductive layer may be TiOPc or other materials whose conductivity increases due to illumination.

[0061] The fluid control system 2 is used to control the injection of the liquid to be tested into the fluid channel of the optoelectronic chip 4 and control the flow rate. The optical system sends a light pattern signal to the optoelectronic chip, and projects a plurality of light patterns 15 on the optoelectronic chip, such as a light array (such as Figure 3 or Figure 4 As shown). At least one excitation light source is provided in the optical system, which can emit excitation light to the light pattern area to excite the fluorescent coded microspheres in the liquid to be tested or the fluorescent molecules on the surface thereof to generate fluorescent signals.

[0062] In some embodiments, the optical system can introduce one or two light patterns and two excitation lights. The light patterns can be used to generate virtual electrodes on the optoelectronic chip to manipulate the coded microspheres. The excitation lights can excite the fluorescence of the coded fluorescent microspheres themselves and the fluorescence of molecules captured on the surface of the microspheres.

[0063] Figure 3 and Figure 4 It is an arrangement of different arrays of light patterns. The circular array is the projected pattern that is reduced by 20 times and then projected onto the optoelectronic chip, so that the size of the light spot on the chip matches the size of the particles to be captured. For example, the size of the coded microsphere is 5 microns, so when designing the light spot array pattern, it is designed to be a light spot array of about 100 microns.

[0064] In some embodiments, the optical system projects the light pattern emitted by the light pattern emission source 6 onto the optoelectronic chip to form a light spot array composed of multiple light spot areas. The conductivity of the photoconductive layer in the light spot area is increased, and electrons can move to its surface to form a virtual electrode. An uneven electric field is formed in the light spot area, so that the fluorescent coded microspheres in the liquid to be tested are moved closer to the light spot array, thereby achieving the positioning of the fluorescent coded microspheres.

[0065] In some embodiments, the surface of the fluorescent coded microspheres is modified so that different molecules can be captured specifically. After the fluorescent coded microspheres are positioned, the first excitation light 7 and the second excitation light 8 are emitted to the light spot array through the optical system to respectively excite the fluorescence of the fluorescent coded microspheres and the molecules captured by the fluorescent coded microspheres; the fluorescence emitted by the fluorescent coded microspheres and their surfaces is collected through the image collection system, and the concentration of the molecule to be measured is obtained by comparing it with the known data.

[0066] The electrical signal control system is used to adjust the electrical signals in the optoelectronic chip, such as voltage and frequency, to control the movement of the fluorescent microspheres. The image processing system is used to collect and process the fluorescent signals emitted by the optoelectronic chip, and can observe the fluorescence of the coded microspheres and the molecules on the surface of the microspheres in real time, identify which microspheres they are, and which molecules are captured.

[0067] In some embodiments, the coded microspheres are made of two fluorescent dyes in different mixing ratios, and the two different fluorescent dyes are mixed according to seven different concentrations, which can reach 49 types.

[0068] The working principle of the present invention is as follows: when in use, an electrical signal is input from the outside, and by adjusting the voltage, current, and waveform of the input electrical signal, and by matching the projected light pattern, an uneven electric field is formed in the fluid channel of the optoelectronic chip. When the fluorescent coded microspheres are in this space, they are affected by the dielectrophoretic force, and then positioned and moved in a horizontal position. Due to the effect of the dielectrophoretic force, the coded microspheres are driven to approach these light spot arrays. After the coded microspheres are positioned, the excitation light of the corresponding wavelength that can excite the coded microspheres and the microsphere surface in the optical system is turned on to generate and identify fluorescent signals.

[0069] The following is further described in conjunction with specific embodiments.

[0070] Example 1

[0071] See also Figure 1 and Figure 2 The photoelectric positioning coded microsphere detection device may include a photoelectric chip, a signal generator (specific electrical signal control system), a group of optical beam reduction lenses (reduced by 20 times), a light pattern control projection system (both of which realize the specific optical system), a camera (CCD) (specific image processing system), and a micropump for controlling liquid injection (specific fluid control system).

[0072] The optoelectronic chip uses ITO conductive glass as a transparent electrode, including an upper ITO electrode and a lower ITO electrode. The ITO electrodes are respectively arranged on a glass substrate, and a layer of photoconductive material is coated on the lower ITO layer. The distance between the upper ITO electrode and the lower ITO electrode is maintained at a distance of 100 microns. The area of ​​the ITO electrode is 1cm×1cm. The upper ITO layer and the lower ITO layer are connected to a signal generator. When used, an external signal is input. By adjusting the voltage, current, and waveform of the input signal, and matching the projected light pattern, an uneven electric field is formed in the space between the electrodes. When the nanospheres are in this space, they are affected by the dielectrophoretic force, and then positioned in the horizontal position.

[0073] After being illuminated, the conductivity of the photoconductive material increases, and the charge is transferred from the ITO glass to the photoconductive layer. The illuminated area is like a virtual electrode. In other words, the light pattern is in the form of an electrode with a variable shape. When the array light pattern is shone on the photoconductive material, the light pattern above becomes conductive. Due to the effect of the dielectrophoretic force, the coded microspheres are driven to approach these light spot arrays. After the coded microspheres are positioned, the lasers with corresponding wavelengths that excite the coded microspheres and the microsphere surface are turned on. For example, the laser that excites the coded microsphere particles has a wavelength of 638nm, and the laser that excites the molecules on the surface of the coded microsphere particles has a wavelength of 532nm. The fluorescence color of the particles and the fluorescence color of the surface are captured by an optical camera. The two fluorescence signals are combined with each other and matched with the known data (how much brightness corresponds to how much concentration), so that the concentration of the molecule to be measured can be obtained.

[0074] After measuring a batch of data, turn off the voltage and current of the signal generator, turn off the illumination pattern, turn off the 532nm and 638nm lights, use an automatic micropump to fill in the liquid, rinse out this batch of solution, load the next batch of solution containing the coded microspheres to be tested, and then measure the next batch of data.

[0075] Example 2

[0076] A real-time fluorescence coded microsphere sorting method, the light pattern used is as follows Figure 8As shown in the figure, the working area of ​​the optoelectronic chip is divided into three parts: the first part is the particle aggregation area and real-time detection area, the second part is the light pattern conversion area, and the third part is the sorting and separation area. Figure 5 , Figure 6 , Figure 7 shown.

[0077] When three types of coded microspheres (for example, numbered (1), (2), and (3)) are mixed in a sample, different molecules will be captured. The coded microspheres with captured molecules on them enter the optoelectronic chip through an automated micropump. Since the coded microspheres are scattered and random, they need to be arranged in a straight line for real-time detection.

[0078] like Figure 5 As shown, virtual electrodes are formed where the light pattern is illuminated. When particles encounter the light pattern, they are pushed toward the center by negative electrophoretic force, thus gathering the scattered particles in the middle and forming a straight line. Then they enter the real-time detection area, where they are illuminated by light of 638nm wavelength to excite the particles. The real-time monitoring system then calibrates which type (1)(2)(3) the incoming coded microspheres belong to.

[0079] like Figure 6 The light pattern conversion area shown is actually equivalent to a light conversion switch. Figure 5 The particle obtained is (1), then we form Figure 6 In the figure (a), the coded microsphere moves upward. Figure 5 The particle obtained is (2), then Figure 6 The light pattern in the image is not projected, allowing the particle to move straight forward. Figure 5 The particle obtained is (3), then we form Figure 6 In the figure (b), the coded microsphere moves downward.

[0080] like Figure 7 In the sorting area shown, there are three light pattern channels, allowing the coded microspheres to enter three different sorting areas, thereby achieving the separation and detection of different coded microspheres and the different target molecules they capture.

[0081] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0082] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A method for sorting fluorescent coded microspheres, characterized in that: The following steps are involved: Mix different coded microspheres in the sample to be tested, and capture the corresponding different target molecules to obtain the test solution; The fluid to be tested is controlled to enter the optoelectronic chip through a fluid control system. The optoelectronic chip includes a first transparent electrode and a second transparent electrode. A fluid channel is formed between the first transparent electrode and the second transparent electrode. A photoconductive layer is arranged on a side of the second transparent electrode close to the fluid channel. The photoconductive layer can change conductivity under light. The optoelectronic chip is connected to an electrical signal control system. The fluorescent coded microspheres can be gathered and positioned in the light pattern irradiation area. The material of the photoconductive layer is TiOPc or hydrogenated amorphous silicon. By projecting a light pattern onto the optoelectronic chip, the optoelectronic chip is divided into a gathering area, a real-time detection area, a light pattern conversion area, and a sorting and separation area in sequence; the gathering area arranges the scattered fluorescent coded microspheres into the real-time detection area; in the real-time detection area, the fluorescent coded microspheres are arranged linearly, and the fluorescent coded microspheres are irradiated with excitation light and the fluorescence is collected to identify the fluorescent coded microspheres; in the light pattern conversion area, a corresponding light pattern channel is generated according to the fluorescent coded microsphere information obtained in the real-time detection area, and the fluorescent coded microspheres are guided to the corresponding sorting and separation area; the sorting and separation area includes a plurality of flow channels to separate and export the fluorescent coded microspheres, and the fluorescent coded microspheres contain iron-containing nanoparticles that can be manipulated by an external magnetic field; The liquid to be tested passes through the aggregation area, the real-time detection area, the light pattern conversion area, and the sorting and separation area in sequence to achieve the sorting of the fluorescent coded microspheres and the corresponding molecules to be tested.

2. The sorting method according to claim 1, characterized in that: The distance between the first transparent electrode and the second transparent electrode is 50-200 micrometers.

3. The sorting method according to claim 1, characterized in that: The first transparent electrode and the second transparent electrode are ITO glass.

4. The sorting method according to claim 1, characterized in that: The fluorescent coding microspheres are made of two fluorescent dyes in different mixing ratios.

5. The sorting method according to claim 1, characterized in that: The surface of the fluorescent coding microsphere is modified so as to be able to specifically capture different molecules.

6. The sorting method according to claim 1, characterized in that: The gathering area includes two strip-shaped light patterns, and both of the two strip-shaped light patterns lead to the real-time detection area.

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