A method and device for analyzing whole blood stock solution cells

Abstract

The present invention discloses a method and device for analyzing cells in whole blood stock solution, belonging to the field of cell analysis. The whole blood stock solution is injected into an offset groove of a microfluidic chip; when the cells move, they encounter an obstacle column, and due to the flow resistance between the cells and the obstacle column, the cells migrate to a preset position in the offset groove; the target cells enter a focusing groove through a first filtering hole in a cell filter plate, and the plasma enters the focusing groove through a second filtering hole, and the plasma is located on both sides of the target cells; the remaining part continues to move along the offset groove and flows out from a first liquid outlet; the plasma pushes the target cells to move along the focusing groove to form a single-cell flow; the single-cell flow is optically detected; the detected target cells and plasma flow out from a second liquid outlet. Through the above steps, no additional reagents are used during the formation of the single-cell flow, and the blood can be transfused back into the body after analysis; a pure physical structure is adopted during the formation of the single-cell flow, and problems such as background noise caused by ultrasonic focusing will not occur.

Description

Technical Field

[0001] The present invention relates to the field of cytometers, and more particularly to a method and apparatus for analyzing whole blood stock solution cells. Background Art

[0002] Flow cytometry is a fundamental and versatile tool for cell quantification, biochemical analysis, and cell sorting. Traditionally known as in vitro flow cytometry, it has been widely used in basic biomedical research and clinical practice. In this technique, target cells are obtained from patients or experimental animals, labeled with fluorescent tracers, and then introduced into a sheath fluid stream. Upon excitation by a laser beam in the stream, the emitted fluorescence is detected and analyzed to provide key physical and biochemical information. However, traditional flow cytometry is limited to in vitro analysis and requires blood extraction and processing, which alters the natural biological environment of blood cells. In addition, the limited blood sample volume in small animal models such as rats and rabbits can significantly affect their physiological state through multiple long-term blood draws. To address these limitations, in vivo flow cytometry has been proposed in recent years.

[0003] The basic principle of in vivo flow cytometry involves using plasma as the sheath fluid stream, eliminating the need for additional reagents. When cells flow through capillaries or drainage tubes and pass through a laser slit, exogenous or endogenous chemical contrast agents are excited to generate fluorescence signals that provide cell information. Compared with in vitro flow cytometry, in vivo flow cytometers can perform real-time, non-invasive, and dynamic monitoring of cells in their natural environment. Currently, due to technical limitations, in vivo flow cytometry is mainly used to detect blood in capillaries of organs such as the ear and retina. However, the slow blood flow velocity and long systemic circulation time in these organs limit the amount of blood tests that can be completed in a short time. Alternatively, directly drawing blood from veins or arteries and forming a "single cell" stream by methods such as ultrasound or inertial focusing for flow cytometry detection, although statistically significant, faces challenges such as cell aggregation and high red blood cell count, which introduce background noise and hinder single cell resolution analysis. Summary of the Invention

[0004] To overcome the deficiencies of the prior art, one object of the present invention is to provide a method for analyzing whole blood stock solution cells that directly uses whole blood stock solution without the use of additional reagents and can be reinfused into the human body or animal body after blood analysis.

[0005] To overcome the deficiencies of the prior art, another object of the present invention is to provide an apparatus for analyzing whole blood stock solution cells that directly uses whole blood stock solution without the use of additional reagents and can be reinfused into the human body or animal body after blood analysis.

[0006] One object of the present invention is achieved by adopting the following technical solution:

[0007] A whole blood stock solution cell analysis method comprises the following steps:

[0008] Injecting whole blood stock solution: injecting the whole blood stock solution into the offset slot of the microfluidic chip;

[0009] Cell deviation: the whole blood stock solution moves in the deviation groove along the length direction of the microfluidic chip. The cells in the whole blood stock solution encounter obstacle columns during the movement. Through the flow resistance between the cells and the obstacle columns, the cells migrate to the preset position in the deviation groove;

[0010] Filtering target cells and plasma: target cells in the whole blood stock solution enter the focusing tank through the first filter hole in the cell filter plate, and plasma in the whole blood stock solution enters the focusing tank through the second filter hole, and the plasma is located on both sides of the target cells;

[0011] The remaining part flows out: the remaining part of the whole blood stock solution continues to move along the offset groove and flows out from the first liquid outlet;

[0012] Forming a single cell stream: The plasma pushes the target cells to move along the focusing groove, the width of the focusing groove shrinks, and the target cells are focused to form a single cell stream;

[0013] Optical detection: optical detection of single cell flow;

[0014] Target cells and plasma outflow: The detected target cells and plasma flow out from the second liquid outlet.

[0015] Furthermore, the whole blood stock solution cell analysis method also includes a collecting and mixing step, which specifically includes: collecting the remaining part of the whole blood stock solution flowing out of the first liquid outlet and the target cells and plasma after detection, and pooling the remaining part of the whole blood stock solution and the target cells and plasma after detection so as to facilitate re-infusion into the body.

[0016] Furthermore, in the cell offset step, the cells are offset to the midline of the microfluidic chip, the first filter hole is arranged at the midline of the microfluidic chip, the offset groove is located above the cell filter plate, and the focusing groove is located below the cell filter plate.

[0017] The second object of the present invention is achieved by adopting the following technical solution:

[0018] A whole blood stock solution cell analysis device for implementing any of the above whole blood stock solution cell analysis methods. The whole blood stock solution cell analysis device includes a microfluidic chip, which includes a cell deflection plate, a cell filtration plate, and a cell focusing plate. The cell deflection plate, the cell filtration plate, and the cell focusing plate are fixedly connected. The cell filtration plate is located between the cell deflection plate and the cell focusing plate. An offset groove is formed between the cell deflection plate and the cell filtration plate, and an obstacle post is provided in the offset groove. A focusing groove is formed between the cell focusing plate and the cell filtration plate. The cell filtration plate is provided with a plurality of first filtration holes and a plurality of second filtration holes. The diameter of the first filtration holes is larger than that of the second filtration holes. The first filtration holes are used for filtering target cells, and the second filtration holes are used for filtering plasma. A plurality of the second filtration holes are located on both sides of the first filtration holes. The focusing groove includes a receiving section, a focusing section, and a straight section. The width of the receiving section is larger than that of the straight section. The focusing section connects the receiving section and the straight section, and the width of the focusing section gradually decreases. The microfluidic chip is further provided with a liquid inlet, a first liquid outlet, and a second liquid outlet. The liquid inlet and the first liquid outlet are respectively communicated with both ends of the offset groove, and the second liquid outlet is communicated with the straight section of the focusing groove.

[0019] Further, the number of the obstacle posts is multiple, and the multiple obstacle posts are arranged in a staggered manner.

[0020] Further, the height of the obstacle post is the same as the depth of the offset groove. The diameter of the obstacle post is 5 - 20 microns, and the distance between the obstacle posts is 30 - 100 microns.

[0021] Further, a plurality of the first filtration holes are located on the same straight line.

[0022] Further, the diameter of the first filtration holes is 7 - 40 microns.

[0023] Further, the diameter of the second filtration holes is 1 - 5 microns.

[0024] Further, the whole blood stock solution cell analysis device further includes a light source, an optical lens group, a first photodetector, and a second photodetector. The light emitted by the light source is irradiated to the end of the straight section through the optical lens group to excite the target cells, and the first photodetector and the second photodetector collect scattered light signals and fluorescence signals.

[0025] Further, when the cell deflection plate, the cell filtration plate, and the cell focusing plate are made of a light-transmitting material, the light source is located on one side of the microfluidic chip, and the first photodetector and the second photodetector are located on the other side of the microfluidic chip.

[0026] Furthermore, when the cell offset plate and the cell filtration plate are made of non-light-transmitting materials, and the cell focusing plate is made of a light-transmitting material, the light source, the first photodetector, and the second photodetector are located on one side of the microfluidic chip.

[0027] Compared with the prior art, the method for analyzing whole blood cell suspension of the present invention passes the whole blood cell suspension through the offset groove of the microfluidic chip; the whole blood cell suspension moves in the offset groove along the length direction of the microfluidic chip, and the cells in the whole blood cell suspension encounter the obstacle posts during the movement. Through the flow resistance between the cells and the obstacle posts, the cells migrate to a preset position in the offset groove; the target cells in the whole blood cell suspension enter the focusing groove through the first filtration holes in the cell filtration plate, and the plasma in the whole blood cell suspension enters the focusing groove through the second filtration holes, and the plasma is located on both sides of the target cells; the remaining part in the whole blood cell suspension continues to move along the offset groove and flows out from the first liquid outlet; the plasma pushes the target cells to move along the focusing groove, the width of the focusing groove converges, and the target cells are focused to form a single-cell flow; the single-cell flow is optically detected; the detected target cells and plasma flow out from the second liquid outlet. Through the above steps, no additional reagents are required during the formation of the single-cell flow, and the blood can be reinfused into the human body or animal body after analysis; the separation and dilution steps during the formation of the single-cell flow adopt a pure physical structure, and problems such as background noise caused by ultrasonic focusing in the prior art will not occur. Description of the Drawings

[0028] Figure 1 is a flowchart of the method for analyzing whole blood cell suspension of the present invention;

[0029] Figure 2 is a perspective view of the microfluidic chip of the whole blood cell suspension analysis device of the present invention;

[0030] Figure 3 is Figure 2 an exploded view of the microfluidic chip;

[0031] Figure 4 is Figure 2 another perspective exploded view of the microfluidic chip;

[0032] Figure 5 is Figure 2 a cross-sectional perspective view of the microfluidic chip;

[0033] Figure 6 is Figure 2 a schematic diagram of the working state of the microfluidic chip;

[0034] Figure 7 is Figure 2 a schematic diagram of the working state of the cell offset plate of the microfluidic chip;

[0035] Figure 8 It is a distribution diagram of the number of cells when there are no obstacle columns on the microfluidic chip;

[0036] Figure 9 It is a distribution diagram of the number of cells when there are obstacle columns on the microfluidic chip;

[0037] Figure 10 It is Figure 2 a schematic diagram of the working state of the cell filter plate of the microfluidic chip of

[0038] Figure 11 It is Figure 2 a schematic diagram of the working state of the cell focusing plate of the microfluidic chip of

[0039] Figure 12 It is a schematic diagram of the working state of the first embodiment of the whole blood stock solution cell analysis device of the present invention;

[0040] Figure 13 It is a schematic diagram of the working state of the second embodiment of the whole blood stock solution cell analysis device of the present invention.

[0041] In the figure: 10, microfluidic chip; 11, cell offset plate; 110, liquid inlet; 111, first liquid outlet; 112, second liquid outlet; 113, offset groove; 114, obstacle column; 12, cell filter plate; 120, first filter hole; 121, second filter hole; 122, through hole; 13, cell focusing plate; 130, focusing groove; 131, receiving section; 132, focusing section; 133, straight section; 14, first joint; 15, second joint; 16, third joint; 20, light source; 30, optical mirror group; 40, first photodetector; 50, second photodetector; 60, first dichroic mirror; 70, second dichroic mirror. Specific embodiments

[0042] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0043] It should be noted that when a component is referred to as "fixed to" another component, it can be directly on the other component or there may also be another intermediate component through which it is fixed. When a component is considered to be "connected to" another component, it can be directly connected to the other component or there may also be another intermediate component at the same time. When a component is considered to be "disposed on" another component, it can be directly disposed on the other component or there may also be another intermediate component at the same time. The terms "vertical", "horizontal", "left", "right" and similar expressions used herein are for illustrative purposes only.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which this invention belongs. The terms used in the description of the present invention herein are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0045] Please refer to Figure 1 , the method for analyzing cells in whole blood stock solution of the present invention includes the following steps:

[0046] Injecting whole blood stock solution: Inject the whole blood stock solution through the offset groove 113 of the microfluidic chip 10;

[0047] Cell offset: The whole blood stock solution moves in the offset groove 113 along the length direction of the microfluidic chip 10. The cells in the whole blood stock solution encounter the obstacle posts 114 during the movement. Through the flow resistance between the cells and the obstacle posts 114, the cells migrate to a preset position in the offset groove 113;

[0048] Filtering target cells and plasma: The target cells in the whole blood stock solution enter the focusing groove 130 through the first filtering holes 120 in the cell filtering plate 12, and the plasma in the whole blood stock solution enters the focusing groove 130 through the second filtering holes 121. The plasma is located on both sides of the target cells;

[0049] Outflow of the remaining part: The remaining part in the whole blood stock solution continues to move along the offset groove 113 and flows out from the first liquid outlet 111;

[0050] Forming a single-cell flow: The plasma pushes the target cells to move along the focusing groove 130. The width of the focusing groove 130 converges, and the target cells are focused to form a single-cell flow;

[0051] Optical detection: Optical detection is performed on the single-cell flow;

[0052] Outflow of target cells and plasma: The detected target cells and plasma flow out from the second liquid outlet 112.

[0053] Specifically, in the step of injecting the whole blood stock solution, the microfluidic chip 10 is provided with a liquid inlet 110. The liquid inlet 110 is located at the end of the offset groove 113 and is in communication with the outside. The whole blood stock solution is the blood extracted from the human body or animal body. The whole blood stock solution includes plasma, target cells and other cells. In this embodiment, the target cells are white blood cells and red blood cells.

[0054] During the cell offset process, the preset position of the offset groove 113 is the central area of the offset groove 113. The purpose of the cells migrating to the preset position in the offset groove 113 is to enable more target cells to pass through the first filtration holes 120 and filter into the focusing groove 130 during the filtration step, improve the filtration efficiency of the target cells, and prevent the target cells from flowing out of the microfluidic chip 10 along the first liquid outlet 111 without entering the focusing groove 130.

[0055] In the step of filtering the target cells and plasma, the diameter of the first filtration holes 120 is larger than that of the second filtration holes 121, so that the target cells enter the focusing groove 130 along the first filtration holes 120, and the plasma enters the focusing groove 130 along the second filtration holes 121. The second filtration holes 121 are located on both sides of the first filtration holes 120, so that the plasma entering the focusing groove 130 is located on both sides of the target cells. The plasma on both sides pushes the target cells to move, and cooperates with the shape of the focusing groove 130 to form a single-cell flow of the target cells. The offset groove 113 is located above the cell filter plate 12, and the focusing groove 130 is located below the cell filter plate 12. So that the target cells and plasma are automatically filtered under the action of gravity.

[0056] In the step of flowing out the remaining part, a large number of cells in the whole blood cells randomly flow out through the first liquid outlet 111, which plays a role in diluting the whole blood stock solution, and the cells flowing out from the first liquid outlet 111 can be reused and input into the human body or animal body.

[0057] In the step of forming a single-cell flow, the plasma pushes the target cells to move along the focusing groove 130. The receiving section 131 of the focusing groove 130 receives the filtered target cells and plasma. The width of the focusing section 132 converges, so that the target cells are focused to form a single-cell flow; the straight section 133 enables the single-cell flow to move in a straight line, and a detection area is formed at the end of the straight section 133 to detect the single-cell flow.

[0058] In the optical detection step, the light emitted by the light source 20 passes through the optical lens group 30 and irradiates to the detection area to form an excitation light path, a shaped light spot is formed in the detection area, the target cells are excited to generate scattered light signals or fluorescence signals, and the first photodetector 40 and the second photodetector 50 collect the scattered light signals or fluorescence signals and perform analysis to realize flow cytometry detection.

[0059] In the target cell and plasma outflow step: The detected target cells and plasma flow out from the second liquid outlet 112 for reuse, or for input into the human body or an animal body.

[0060] The whole blood stock solution cell analysis method further includes a collection and mixing step, specifically: collecting the remaining part of the whole blood stock solution flowing out from the first liquid outlet 111, the detected target cells, and plasma, and pooling the remaining part of the whole blood stock solution, the detected target cells, and plasma for re-infusion into the body.

[0061] Please continue to refer to Figures 2 to 13 , this application also discloses a whole blood stock solution cell analysis device for implementing the above whole blood stock solution cell analysis method. The whole blood stock solution cell analysis device includes a microfluidic chip 10, a light source 20, an optical lens group 30, a first photodetector 40, and a second photodetector 50.

[0062] Please continue to refer to Figure 2 , the microfluidic chip 10 includes a cell offset plate 11, a cell filter plate 12, a cell focusing plate 13, a first connector 14, a second connector 15, and a third connector 16. The cell offset plate 11, the cell filter plate 12, and the cell focusing plate 13 are made of micron-scale materials such as glass, silicon wafers, PDMS, engineering plastics, etc. The cell focusing plate 13 is processed with materials having good optical transparency such as quartz or other glass. The cell offset plate 11, the cell filter plate 12, and the cell focusing plate 13 are connected by curing glue connection or molecular bond connection. The first connector 14, the second connector 15, and the third connector 16 are made of stainless steel, titanium alloy, or other engineering plastic materials with good biocompatibility.

[0063] Please continue to refer to Figure 3 , Figure 4 and Figures 7 to 9 , the cell offset plate 11 is used to offset the cells in the whole blood stock solution so that the target cells can be exactly filtered into the focusing groove 130.

[0064] The cell offset plate 11 is provided with a liquid inlet 110, a first liquid outlet 111, and a second liquid outlet 112. The liquid inlet 110 is located at one end of the cell offset plate 11, and the first liquid outlet 111 and the second liquid outlet 112 are located at the other end of the cell offset plate 11. The liquid inlet 110 is used to install the first connector 14 so that the whole blood stock solution can be injected into the offset groove 113. The first liquid outlet 111 is used to install the second connector 15 for outputting the remaining liquid after filtering the target cells and plasma, so as to facilitate subsequent recycling. The second liquid outlet 112 is used to install the third connector 16 for outputting the analyzed target cells and plasma, so as to facilitate subsequent recycling. Specifically, the liquid inlet 110 and the first liquid outlet 111 are communicated with the offset groove 113 and are located at both ends of the offset groove 113, and the second liquid outlet 112 is communicated with the focusing groove 130.

[0065] On the side of the cell offset plate 11 facing the cell filter plate 12, there is also an offset groove 113. The depth of the offset groove 113 is 20 - 100 microns. A plurality of obstacle columns 114 are provided in the offset groove 113, and the plurality of obstacle columns 114 are etched out of position at equal distances. The diameter of the obstacle column 114 is 5 - 20 microns, and the distance between the obstacle columns 114 is 30 - 100 microns. The cell offset plate 11 utilizes the flow resistance between the cells and the obstacle columns 114 to complete the migration of the cells to the middle position of the flow channel. That is, in the width direction (y-direction) of the flow channel groove, the number of cells in the middle part of the groove is significantly more than that in the two sides of the groove, as Figure 7 shown. If there are no obstacle columns 114 in the groove of the cell offset plate 11, the cell number distribution along the width direction (y-direction) of the flow channel is as Figure 8 shown. When there are obstacle columns 114 in the groove of the cell offset plate 11, the cell number distribution along the width direction (y-direction) of the flow channel is as Figure 9 shown.

[0066] Please continue to refer to Figure 4 and Figure 10 shown. The cell filter plate 12 is provided with a first filter hole 120, a second filter hole 121, and a through hole 122. The first filter hole 120 is used for filtering target cells, the second filter hole 121 is used for plasma filtration, and the through hole 122 is used for collecting the analyzed plasma and target cells. The diameter of the first filter hole 120 is larger than that of the second filter hole 121. A plurality of second filter holes 121 are located on both sides of the first filter hole 120. Specifically, a plurality of first filter holes 120 are arranged in a straight line, a plurality of second filter holes 121 are located on both sides of the straight line, and the plurality of second filter holes 121 are evenly distributed. The aperture of the first filter hole 120 is 7 - 40 microns, and the aperture of the second filter holes 121 distributed on both sides is 1 - 5 microns.

[0067] Please continue to refer to Figure 3 and Figure 11As shown, the cell focusing plate 13 is provided with a focusing groove 130. The focusing groove 130 is provided with a receiving section 131, a focusing section 132, and a straight section 133. The focusing section 132 connects the receiving section 131 and the straight section 133. The widths of the receiving section 131 and the straight section 133 are fixed, and the width of the receiving section 131 is greater than the width of the straight section 133. The width of the focusing section 132 gradually decreases from the receiving section 131 to the straight section 133, and the focusing of target cells is achieved through a converging structure. The depth of the focusing groove 130 is 10 - 100 microns, and the width of the straight section 133 is 10 - 200 microns. Biological particles such as cells, under the action of plasma, achieve hydrodynamic focusing at the position of the straight section 133.

[0068] Please continue to refer to Figure 12 , in the first embodiment of the whole blood stock solution cell analysis device, the cell offset plate 11, the cell filter plate 12, and the cell focusing plate 13 are made of light-transmitting materials. The light source 20 is located on one side of the microfluidic chip 10, and the first photodetector 40 and the second photodetector 50 are located on the other side of the microfluidic chip 10. The light of the light source 20 passes through the optical mirror group 30 to excite a shaped light spot in the detection area of the straight section 133 of the microfluidic chip 10. When biological particles such as cells after hydrodynamic focusing pass through the excited shaped light spot, scattered light signals or fluorescence signals will be generated. The first photodetector 40 and the second photodetector 50 receive the scattered light signals or fluorescence signals and analyze the light signals.

[0069] Please continue to refer to Figure 13 , in the second embodiment of the whole blood stock solution cell analysis device, the cell offset plate 11 and the cell filter plate 12 are made of light-impermeable materials, and the cell focusing plate 13 is made of light-transmitting materials. The light source 20, the optical mirror group 30, the first photodetector 40, the second photodetector 50, the first dichroic mirror 60, and the second dichroic mirror 70 are located on the same side of the microfluidic chip 10, and the epi-illumination method is used for flow cytometry detection. The light of the light source 20 passes through the optical mirror group 30 and the first dichroic mirror 60 and irradiates to the detection area to excite a shaped light spot. When biological particles such as cells after hydrodynamic focusing pass through the excited shaped light spot, scattered light signals or fluorescence signals will be generated. The scattered light signals or fluorescence signals are transmitted through the second dichroic mirror 70 to the first photodetector 40 and the second photodetector 50 to receive the scattered light signals or fluorescence signals and analyze the light signals.

[0070] When using the whole blood stock solution cell analysis device, as Figure 6As shown, the whole blood stock solution is injected into the offset groove 113 of the microfluidic chip 10 through the first connector 14; the whole blood stock solution moves in the offset groove 113 along the length direction of the microfluidic chip 10. During the movement, the cells in the whole blood stock solution encounter the obstacle posts 114, and due to the flow resistance between the cells and the obstacle posts 114, the cells migrate to a preset position in the offset groove 113; the target cells in the whole blood stock solution enter the focusing groove 130 through the first filtration holes 120 in the cell filtration plate 12, and the plasma in the whole blood stock solution enters the focusing groove 130 through the second filtration holes 121. The plasma is located on both sides of the target cells. The remaining part of the whole blood stock solution continues to move along the offset groove 113 and flows out from the first liquid outlet 111. The plasma pushes the target cells to move along the focusing groove 130. The width of the focusing groove 130 converges, and the target cells are focused to form a single-cell flow, and the single-cell flow is subjected to optical detection; the detected target cells and plasma flow out from the second liquid outlet 112.

[0071] During the process of forming a single-cell flow by the whole blood stock solution cell analysis device, no additional reagents are required, and the blood can be transfused back into the human body or animal body after analysis; during the process of forming a single-cell flow, steps such as separation and dilution adopt a pure physical structure, and problems such as background noise caused by ultrasonic focusing in the prior art will not occur.

[0072] The above embodiments only represent several implementation manners of the present invention, and the description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can be made. These are all equivalent modifications and evolutions made to the above embodiments based on the essence of the present invention, and these all belong to the protection scope of the present invention.

Claims

1. A whole blood stock solution cell analysis method, characterized in that: The following steps are involved: Injecting whole blood stock solution: injecting the whole blood stock solution into the offset slot of the microfluidic chip; Cell deviation: the whole blood stock solution moves in the deviation groove along the length direction of the microfluidic chip. The cells in the whole blood stock solution encounter obstacle columns during the movement. Through the flow resistance between the cells and the obstacle columns, the cells migrate to the preset position in the deviation groove; Filtering target cells and plasma: target cells in the whole blood stock solution enter the focusing tank through the first filter hole in the cell filter plate, and plasma in the whole blood stock solution enters the focusing tank through the second filter hole, and the plasma is located on both sides of the target cells. The diameter of the first filter hole is greater than the diameter of the second filter hole. A plurality of the first filter holes are arranged in a straight line, and a plurality of the second filter holes are located on both sides of the first filter hole. The remaining part flows out: the remaining part of the whole blood stock solution continues to move along the offset groove and flows out from the first liquid outlet; Forming a single cell stream: The plasma pushes the target cells to move along the focusing groove, the width of the focusing groove shrinks, and the target cells are focused to form a single cell stream; Optical detection: optical detection of single cell flow; Target cells and plasma flow out: The detected target cells and plasma flow out from the second liquid outlet; Collecting and mixing: collecting the remaining part of the whole blood stock solution flowing out of the first liquid outlet and the target cells and plasma after detection, and pooling the remaining part of the whole blood stock solution and the target cells and plasma after detection so as to be re-infused into the body.

2. The whole blood stock solution cell analysis method according to claim 1, characterized in that: In the cell shifting step, the cells are shifted to the midline of the microfluidic chip, the first filter hole is arranged at the midline of the microfluidic chip, the shifting groove is located above the cell filter plate, and the focusing groove is located below the cell filter plate.

3. A whole blood stock solution cell analysis device, used to implement the whole blood stock solution cell analysis method according to any one of claims 1 to 2, the whole blood stock solution cell analysis device comprising a microfluidic chip, characterized in that: The microfluidic chip includes a cell offset plate, a cell filter plate and a cell focusing plate, wherein the cell offset plate, the cell filter plate and the cell focusing plate are fixedly connected, the cell filter plate is located between the cell offset plate and the cell focusing plate, an offset groove is formed between the cell offset plate and the cell filter plate, an obstacle column is arranged in the offset groove, a focusing groove is formed between the cell focusing plate and the cell filter plate, the cell filter plate is provided with a plurality of first filter holes and a plurality of second filter holes, the diameter of the first filter hole is greater than the diameter of the second filter hole, the first filter hole is used to filter target cells, the second filter hole is used to filter plasma, a plurality of second filter holes are located on both sides of the first filter hole, the focusing groove includes a receiving section, a focusing section and a straight line section, the width of the receiving section is greater than the width of the straight line section, the focusing section connects the receiving section and the straight line section, and the width of the focusing section gradually decreases, the microfluidic chip is also provided with a liquid inlet, a first liquid outlet and a second liquid outlet, the liquid inlet and the first liquid outlet are respectively connected to the two ends of the offset groove, and the second liquid outlet is connected to the straight line section of the focusing groove.

4. The whole blood stock solution cell analysis device according to claim 3, characterized in that: There are multiple obstacle columns, and the multiple obstacle columns are staggered.

5. The whole blood stock solution cell analysis device according to claim 4, characterized in that: The height of the obstacle column is the same as the depth of the offset groove, the diameter of the obstacle column is 5 to 20 microns, and the spacing between the obstacle columns is 30 to 100 microns.

6. The whole blood stock solution cell analysis device according to claim 3, characterized in that: The plurality of first filter holes are located on the same straight line.

7. The whole blood stock solution cell analysis device according to claim 3, characterized in that: The diameter of the first filter hole is 7-40 microns.

8. The whole blood stock solution cell analysis device according to claim 4, characterized in that: The diameter of the second filter hole is 1-5 microns.

9. The whole blood stock solution cell analysis device according to claim 3, characterized in that: The whole blood stock solution cell analysis device also includes a light source, an optical lens group, a first photodetector and a second photodetector. The light emitted by the light source passes through the optical lens group and irradiates the end of the straight segment to excite the target cells. The first photodetector and the second photodetector collect scattered light signals and fluorescence signals.

10. The whole blood stock solution cell analysis device according to claim 9, characterized in that: When the cell deviation plate, the cell filter plate and the cell focusing plate are made of light-transmitting materials, the light source is located on one side of the microfluidic chip, and the first photodetector and the second photodetector are located on the other side of the microfluidic chip.

11. The whole blood stock solution cell analysis device according to claim 9, characterized in that: When the cell deviation plate and the cell filter plate are made of non-light-transmitting materials, and the cell focusing plate is made of light-transmitting materials, the light source, the first photodetector, and the second photodetector are located on one side of the microfluidic chip.

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