Whole blood stock solution cell analysis method and device

Abstract

The invention discloses a whole blood stock solution cell analysis method and a whole blood stock solution cell analysis device, and belongs to the field of cell analysis. The cells encounter the obstacle column in the moving process, and the cells migrate to the preset position in the offset groove through the flow resistance between the cells and the obstacle column; target cells enter the focusing groove through first filtering holes in the cell filtering plate, plasma enters the focusing groove through second filtering holes, and the plasma is located on the two sides of the target cells; the rest part continuously moves along the offset groove and flows out from the first liquid outlet; the plasma pushes the target cells to move and focus along the focusing groove to form single cell flow; performing optical detection on the single cell flow; the detected target cells and plasma flow out of the second liquid outlet, through the steps, no extra reagent is needed in the single cell flow forming process, and blood can be infused into the body again after being analyzed; a pure physical structure is adopted in the single cell flow forming process, and the problems of background noise and the like caused by ultrasonic focusing cannot be generated.

Description

Technical Field

[0001] The present invention relates to the field of cytometers, and in particular to a whole blood stock solution cell analysis method and device. Background Art

[0002] Flow cytometry is a fundamental and versatile tool for cell quantification, biochemical analysis, and cell sorting. Traditionally referred to 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 flow, the emitted fluorescence is detected and analyzed to provide critical physical and biochemical information. However, conventional 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 amount of blood samples in small animal models such as rats and rabbits can significantly affect their physiological status 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 the use of plasma as a sheath fluid stream, eliminating the need for additional reagents. As cells flow through capillaries or drainage tubes and pass through the laser slit, exogenous or endogenous chemical contrast agents are excited to produce fluorescent signals that provide cellular information. Compared with in vitro flow cytometry, in vivo flow cytometry allows 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 the capillaries of organs such as the ear and retina. However, the slow blood flow rate and long systemic circulation time in these organs limit the amount of blood testing that can be completed in a short period of time. Alternatively, blood is drawn directly from veins or arteries and a "single cell" stream is formed for flow cytometry detection through methods such as ultrasound or inertial focusing. Although statistically significant, it faces challenges such as cell aggregation and high proportion of red blood cell counts, which introduce background noise and hinder single-cell resolution analysis. Summary of the invention

[0004] In order to overcome the shortcomings of the prior art, one of the objectives of the present invention is to provide a whole blood stock solution cell analysis method that directly uses whole blood stock solution without using additional reagents and can be re-injected into the human body or animal body after blood analysis.

[0005] In order to overcome the deficiencies of the prior art, the second object of the present invention is to provide a whole blood stock solution cell analysis device that directly uses whole blood stock solution without the need for additional reagents and can be reintroduced into the human body or animal body after blood analysis.

[0006] One of the purposes of the present invention is achieved by the following technical solution: A whole blood stock solution cell analysis method comprises the following steps: 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 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 outflow: The detected target cells and plasma flow out from the second liquid outlet.

[0007] 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.

[0008] 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.

[0009] The second object of the present invention is achieved by adopting the following technical solution: A whole blood stock solution cell analysis device, used to implement any of the above-mentioned whole blood stock solution cell analysis methods, the whole blood stock solution cell analysis device comprises a microfluidic chip, the microfluidic chip comprises a cell offset plate, a cell filter plate and a cell focusing plate, 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, and the second filter hole is used to filter plasma. A plurality of the 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 section. The width of the receiving section is greater than the width of the straight section. The focusing section connects the receiving section and the straight section. 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 section of the focusing groove.

[0010] Furthermore, there are multiple obstacle columns, and the multiple obstacle columns are staggered.

[0011] Furthermore, 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.

[0012] Furthermore, a plurality of the first filter holes are located on the same straight line.

[0013] Furthermore, the diameter of the first filter hole is 7-40 microns.

[0014] Furthermore, the diameter of the second filter hole is 1-5 microns.

[0015] Furthermore, 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 to 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.

[0016] Furthermore, 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.

[0017] Furthermore, 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.

[0018] Compared with the prior art, the whole blood stock solution cell analysis method of the present invention injects the whole blood stock solution into the offset groove of the microfluidic chip; the whole blood stock solution moves in the offset groove along the length direction of the microfluidic chip, and 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 offset groove; the target cells in the whole blood stock solution enter the focusing groove through the first filter hole in the cell filter plate, and the plasma in the whole blood stock solution enters the focusing groove through the second filter hole, and 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 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 is narrowed, 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 needed in the process of forming the single cell flow, and the blood can be re-injected into the human body or animal body after analysis; the separation and dilution steps in the process of forming the single cell flow adopt a purely physical structure, and will not produce problems such as background noise caused by ultrasonic focusing in the prior art. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a flow chart of the whole blood stock solution cell analysis method of the present invention; Figure 2 A three-dimensional diagram of a microfluidic chip of a whole blood stock solution cell analysis device of the present invention; Figure 3 for Figure 2 An exploded view of a microfluidic chip; Figure 4 for Figure 2 Another perspective exploded view of the microfluidic chip; Figure 5 for Figure 2 A three-dimensional cross-sectional view of a microfluidic chip; Figure 6 for Figure 2 Schematic diagram of the use status of the microfluidic chip; Figure 7 for Figure 2 Schematic diagram of the working state of the cell displacement plate of the microfluidic chip; Figure 8 This is the cell number distribution diagram when there is no obstacle column in the microfluidic chip; Fig. 9 This is the cell number distribution diagram when the microfluidic chip has an obstacle column; Fig.10 for Figure 2 Schematic diagram of the working state of the cell filter plate of the microfluidic chip; Fig.11 for Figure 2 Schematic diagram of the working state of the cell focusing plate of the microfluidic chip; Fig.12 It is a working schematic diagram of the first embodiment of the whole blood stock solution cell analysis device of the present invention; Fig.13 FIG. 1 is a schematic diagram of the operation of the second embodiment of the whole blood stock solution cell analysis device of the present invention.

[0020] In the figure: 10, microfluidic chip; 11, cell offset plate; 110, liquid inlet; 111, first liquid outlet; 112, second liquid outlet; 113, offset slot; 114, obstacle column; 12, cell filter plate; 120, first filter hole; 121, second filter hole; 122, through hole; 13, cell focusing plate; 130, focusing slot; 131, receiving section; 132, focusing section; 133, straight line section; 14, first joint; 15, second joint; 16, third joint; 20, light source; 30, optical lens group; 40, first photodetector; 50, second photodetector; 60, first dichroic mirror; 70, second dichroic mirror. DETAILED DESCRIPTION

[0021] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0022] It should be noted that when a component is referred to as being "fixed to" another component, it may be directly on the other component or there may be another intermediate component through which it is fixed. When a component is considered to be "connected to" another component, it may be directly connected to the other component or there may be another intermediate component at the same time. When a component is considered to be "set on" another component, it may be directly set on the other component or there may be another intermediate component at the same time. The terms "vertical", "horizontal", "left", "right" and similar expressions used herein are for illustrative purposes only.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. 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 of the related listed items.

[0024] See also Figure 1 The whole blood stock solution cell analysis method of the present invention comprises the following steps: Injecting whole blood stock solution: injecting the whole blood stock solution through the offset groove 113 of the microfluidic chip 10; Cell deviation: the whole blood stock solution moves in the deviation groove 113 along the length direction of the microfluidic chip 10. The cells in the whole blood stock solution encounter the obstacle column 114 during the movement. Due to the flow resistance between the cells and the obstacle column 114, the cells migrate to the preset position in the deviation groove 113. Filtering target cells and plasma: target cells in the whole blood stock solution enter the focusing tank 130 through the first filtering holes 120 in the cell filtering plate 12, and plasma in the whole blood stock solution enters the focusing tank 130 through the second filtering holes 121, and the plasma is located on both sides of the target cells; The remaining part flows out: 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; 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 is reduced, and the target cells are focused to form a single cell flow; Optical detection: optical detection of single cell flow; Target cells and plasma outflow: The detected target cells and plasma flow out from the second liquid outlet 112 .

[0025] 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 the liquid inlet 110 is connected to the outside. The whole blood stock solution is blood extracted from a human 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.

[0026] During the cell deviation process, the preset position of the deviation groove 113 is the central area of ​​the deviation groove 113. The purpose of the cell migration to the preset position in the deviation groove 113 is to enable more target cells to be filtered into the focusing groove 130 through the first filter hole 120 in the filtering step, thereby improving the filtering efficiency of the target cells and preventing the target cells from flowing out of the microfluidic chip 10 along the first liquid outlet 111 without entering the focusing groove 130.

[0027] In the step of filtering target cells and plasma, the diameter of the first filter hole 120 is larger than the diameter of the second filter hole 121, so that the target cells enter the focusing groove 130 along the first filter hole 120, and the plasma enters the focusing groove 130 along the second filter hole 121. The second filter hole 121 is located on both sides of the first filter hole 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 the target cells form a single cell flow in accordance with the shape of the focusing groove 130. 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. The target cells and plasma are automatically filtered under the action of gravity.

[0028] In the remaining outflow step, a large number of 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 injected into the human body or animal body.

[0029] 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 is reduced to focus the target cells to form a single-cell flow. The straight line segment 133 makes the single-cell flow move along a straight line, and a detection area is formed at the end of the straight line segment 133 to detect the single-cell flow.

[0030] In the optical detection step, the light emitted by the light source 20 passes through the optical lens group 30 to irradiate the detection area to form an excitation light path, forming a shaped light spot in the detection area, and the target cells are excited to generate scattered light signals or fluorescent signals. The first photodetector 40 and the second photodetector 50 collect the scattered light signals or fluorescent signals and analyze them to realize flow cytometry detection.

[0031] In the target cell and plasma outflow step: the detected target cells and plasma flow out from the second liquid outlet 112 so as to be reused and injected into the human body or animal body.

[0032] 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 111 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.

[0033] Please continue reading Figures 2 to 13 The present application also discloses a whole blood stock solution cell analysis device for implementing the above-mentioned 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.

[0034] Please continue reading Figure 2 The microfluidic chip 10 includes a cell displacement plate 11, a cell filter plate 12, a cell focusing plate 13, a first joint 14, a second joint 15 and a third joint 16. The cell displacement plate 11, the cell filter plate 12 and the cell focusing plate 13 are made of micron-grade materials such as glass, silicon wafer, PDMS, engineering plastics and the like. The cell focusing plate 13 is made of materials with good optical transmittance such as quartz or other glass. The cell displacement plate 11, the cell filter plate 12 and the cell focusing plate 13 are connected by curing glue connection or molecular bonding connection. The first joint 14, the second joint 15 and the third joint 16 are made of stainless steel or titanium alloy or other engineering plastic materials with good biocompatibility.

[0035] Please continue reading Figure 3 , Figure 4 as well as Figures 7 to 9 The cell shifting plate 11 is used to shift the cells in the whole blood stock solution so that the target cells can be filtered exactly into the focusing tank 130 .

[0036] The cell deviation 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 deviation plate 11, and the first liquid outlet 111 and the second liquid outlet 112 are located at the other end of the cell deviation 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 deviation 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 for subsequent recycling. The second liquid outlet 112 is used to install the third connector 16 for outputting the target cells and plasma after analysis for subsequent recycling. Specifically, the liquid inlet 110 and the first liquid outlet 111 are connected to the deviation groove 113 and are located at both ends of the deviation groove 113. The second liquid outlet 112 is connected to the focusing groove 130.

[0037] The cell deflection plate 11 is also provided with a deflection groove 113 on the side facing the cell filter plate 12, and the depth of the deflection groove 113 is 20 to 100 microns. A plurality of obstacle columns 114 are provided in the deflection groove 113, and the plurality of obstacle columns 114 are formed by equidistant staggered etching. The diameter of the obstacle column 114 is 5 to 20 microns, and the spacing between the obstacle columns 114 is 30 to 100 microns. The cell deflection plate 11 utilizes the flow resistance between the cells and the obstacle columns 114 to complete the migration of 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 greater than that on the two sides of the groove, such as Figure 7 If there is no obstruction column 114 in the groove of the cell offset plate 11, the number of cells along the width direction (y direction) of the flow channel is distributed as follows: Figure 8 When there is an obstacle column 114 in the groove of the cell deviation plate 11, the number of cells along the width direction (y direction) of the flow channel is distributed as follows: Fig. 9shown.

[0038] Please continue reading Figure 4 as well as Fig.10 As 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 filtering plasma, and the through hole 122 is used for collecting plasma and target cells after analysis. The diameter of the first filter hole 120 is greater than the diameter 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 a plurality of second filter holes 121 are evenly distributed. The aperture of the first filter hole 120 is 7 to 40 microns, and the aperture of the second filter holes 121 distributed on both sides is 1 to 5 microns.

[0039] Please continue reading Figure 3 as well as Fig.11 As shown, the cell focusing plate 13 is provided with a focusing groove 130, and 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 the target cells is achieved by the folding structure. The depth of the focusing groove 130 is 10 to 100 microns, and the width of the straight section 133 is 10 to 200 microns. Under the action of plasma, biological particles such as cells achieve hydrodynamic focusing at the position of the straight section 133.

[0040] Please continue reading Fig.12 In the first embodiment of the whole blood stock solution cell analysis device, the cell deviation 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 lens group 30 to excite the shaped light spot in the detection area of ​​the straight segment 133 of the microfluidic chip 10. When the biological particles such as cells after fluid dynamics focusing pass through the excited shaped light spot, a scattered light signal or a fluorescent signal is generated. The first photodetector 40 and the second photodetector 50 receive the scattered light signal or the fluorescent signal and analyze the light signal.

[0041] Please continue reading Fig.13In 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 opaque materials, and the cell focusing plate 13 is made of translucent materials. The light source 20, the optical lens 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 flow cytometry is performed using an incident illumination method. The light of the light source 20 passes through the optical lens group 30 and the first dichroic mirror 60 to irradiate the detection area to excite the shaping light spot. When the biological particles such as cells after fluid dynamics focusing pass through the excitation shaping light spot, a scattered light signal or a fluorescent signal is generated. The scattered light signal or the fluorescent signal is transmitted to the first photodetector 40 and the second photodetector 50 through the second dichroic mirror 70 to receive the scattered light signal or the fluorescent signal, and the light signal is analyzed.

[0042] When using a whole blood stock cell analysis device, Figure 6 As shown, the whole blood stock solution is injected into the offset groove 113 of the microfluidic chip 10 through the first joint 14; the whole blood stock solution moves in the offset groove 113 along the length direction of the microfluidic chip 10, and the cells in the whole blood stock solution encounter the obstacle column 114 during the movement. Through the flow resistance between the cells and the obstacle column 114, the cells migrate to the preset position in the offset groove 113; the target cells in the whole blood stock solution enter the focusing groove 130 through the first filter hole 120 in the cell filter plate 12, and the plasma in the whole blood stock solution enters the focusing groove 130 through the second filter hole 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 is contracted, and the target cells are focused to form a single cell flow, and the single cell flow is optically detected; the detected target cells and plasma flow out from the second liquid outlet 112.

[0043] No additional reagents are required in the process of forming a single-cell flow by the whole blood stock solution cell analysis device, and the blood can be re-injected into the human body or animal body after analysis; the separation and dilution steps in the process of forming a single-cell flow adopt a purely physical structure, and will not produce problems such as background noise caused by ultrasonic focusing in the prior art.

[0044] The above 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 modifications and improvements can be made without departing from the concept of the present invention, which are equivalent modifications and improvements made to the above embodiments based on the essential technology of the present invention, and all of them 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 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 outflow: The detected target cells and plasma flow out from the second liquid outlet.

2. The whole blood stock solution cell analysis method according to claim 1, characterized in that: 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.

3. 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.

4. A whole blood stock solution cell analysis device, used for implementing the whole blood stock solution cell analysis method according to any one of claims 1 to 3, 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.

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

6. The whole blood stock solution cell analysis device according to claim 5, 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.

7. The whole blood stock solution cell analysis device according to claim 4, characterized in that: The plurality of first filter holes are located on the same straight line.

8. The whole blood stock solution cell analysis device according to claim 4, characterized in that: The diameter of the first filter hole is 7-40 microns.

9. 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.

10. The whole blood stock solution cell analysis device according to claim 4, 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.

11. The whole blood stock solution cell analysis device according to claim 10, 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.

12. The whole blood stock solution cell analysis device according to claim 10, 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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