Cell analyzer and cell analysis method

Abstract

The invention provides a cell analyzer and a cell analysis method.The cell analyzer comprises a chip layer and a runner layer, a liquid inlet, an impedance measuring unit and a liquid driving unit are arranged in the chip layer, the liquid inlet is communicated with a first surface and a second surface of the chip layer, the impedance measuring unit comprises an electrode plate exposed on the first surface of the chip layer, and the liquid driving unit is arranged on the first surface of the chip layer. The liquid driving unit comprises a driving piece exposed on the first surface of the chip layer, the runner layer is combined with the first surface of the chip layer and comprises a runner and a nozzle, the runner is formed in the face, facing the chip layer, of the runner layer, the liquid inlet, the runner and the nozzle are sequentially communicated, and an outlet of the nozzle is formed in the face, away from the chip layer, of the runner layer. At least one part of the electrode plate is located in the flow channel, and at least one part of the driving plate faces the outlet of the nozzle. The cell analyzer provided by the invention can finish accurate quantitative transportation of liquid to be detected and accurate counting of specific cells, realizes accurate measurement of the number and concentration of cells on a chip, and has the characteristics of miniaturization, automation and low cost.

Description

Technical Field

[0001] The invention belongs to the technical field of integrated circuits and relates to a cell analyzer and a cell analysis method. Background Art

[0002] Whole blood cell analysis has a wide range of applications in disease diagnosis and treatment. Almost all diseases are initially tested with whole blood cell testing. Currently, whole blood cell analysis is generally divided into three categories and five categories. The relatively simple three-category whole blood analysis mainly detects blood particles based on impedance measurement and distinguishes different particles based on different signals. Whole blood analyzers based on this method require more complex automated equipment, including pumps, particle separation microstructures, etc. The equipment is large in size and expensive.

[0003] Therefore, how to provide a miniaturized, automated, low-cost cell analyzer and a corresponding cell analysis method has become an important technical problem that needs to be solved urgently by those skilled in the art.

[0004] It should be noted that the above introduction to the technical background is only for the convenience of providing a clear and complete description of the technical solutions of the present application and for the convenience of understanding by those skilled in the art. It cannot be considered that the above technical solutions are well known to those skilled in the art simply because they are described in the background technology section of the present application. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the prior art, an object of the present invention is to provide a cell analyzer and a cell analysis method, which are used to solve the problems of large size and high cost of cell analysis equipment in the prior art.

[0006] To achieve the above-mentioned object and other related objects, the present invention provides a cell analyzer, comprising:

[0007] A chip layer, comprising a first surface and a second surface arranged opposite to each other, wherein a liquid inlet, an impedance measuring unit and a liquid driving unit are provided in the chip layer, wherein the liquid inlet communicates with the first surface and the second surface, the impedance measuring unit comprises an electrode sheet exposed on the first surface, and the liquid driving unit comprises a driving sheet exposed on the first surface;

[0008] A flow channel layer is combined with the first surface of the chip layer, the flow channel layer includes a flow channel and a nozzle, the flow channel is opened on the side of the flow channel layer facing the chip layer, the liquid inlet, the flow channel and the nozzle are connected in sequence, the outlet of the nozzle is opened on the side of the flow channel layer away from the chip layer, at least a part of the electrode sheet is located in the flow channel, and at least a part of the drive sheet faces the outlet of the nozzle.

[0009] Optionally, the liquid driving unit drives the liquid by thermal bubble driving or piezoelectric driving.

[0010] Optionally, there are multiple flow channels, and there are multiple nozzles, and each flow channel is connected to a corresponding nozzle.

[0011] Optionally, at least two of the flow channels have different passage sizes to achieve separation of cells of different particle sizes.

[0012] Optionally, the impedance measurement unit comprises at least two electrode sheets.

[0013] Optionally, the liquid driving unit further includes a driving circuit integrated in the chip layer and electrically connected to the driving chip.

[0014] Optionally, the number of the nozzles is greater than 100.

[0015] Optionally, the liquid to be tested includes blood.

[0016] Optionally, the flow channel layer comprises a photolithographic polymer material.

[0017] The present invention also provides a cell analysis method, comprising the following steps:

[0018] The liquid to be tested is added into the liquid inlet, and a driving plate facing the nozzle outlet is used to drive the liquid to be tested to be ejected from the nozzle and drive the liquid to be tested to flow in the flow channel;

[0019] Using an impedance measurement unit to measure the impedance of cells flowing through the impedance measurement unit in the flow channel and calculate the number of specific cells;

[0020] Calculate the total volume of the liquid to be tested sprayed based on the single spray volume and the number of sprays of a single nozzle;

[0021] The concentration of the specific cells in the liquid to be tested is calculated based on the calculated number of specific cells and the total volume of the liquid to be tested that is ejected.

[0022] As described above, the cell analyzer of the present invention includes a chip layer and a flow channel layer, wherein the chip layer includes a first surface and a second surface arranged oppositely, a liquid inlet, an impedance measurement unit and a liquid drive unit are provided in the chip layer, the liquid inlet is connected to the first surface and the second surface of the chip layer, the impedance measurement unit includes an electrode sheet exposed on the first surface of the chip layer, the liquid drive unit includes a drive sheet exposed on the first surface of the chip layer, the flow channel layer is combined with the first surface of the chip layer and includes a flow channel and a nozzle, the flow channel is opened on the side of the flow channel layer facing the chip layer, the liquid inlet, the flow channel and the nozzle are connected in sequence, the outlet of the nozzle is opened on the side of the flow channel layer away from the chip layer, at least a part of the electrode sheet is located in the flow channel, and at least a part of the drive sheet faces the outlet of the nozzle. The cell analyzer of the present invention can complete the precise quantitative transportation of the liquid to be tested and the accurate counting of specific cells, realize the precise measurement of the number and concentration of cells on the chip, and has the characteristics of miniaturization, automation and low cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 FIG. 1 is a schematic diagram of a cross-sectional structure of a cell analyzer according to an embodiment of the present invention.

[0024] Figure 2 FIG. 4 is a plan view of a cell analyzer according to an embodiment of the present invention.

[0025] Component number description

[0026] 100 Chip Layer

[0027] 101 Liquid inlet

[0028] 102 Impedance measurement unit

[0029] 102a Electrode sheet

[0030] 103 Liquid drive unit

[0031] 103a Driver Plate

[0032] 200 Runner Layer

[0033] 200a First photolithographic polymer material layer

[0034] 200b second photolithographic polymer material layer

[0035] 201 Runner

[0036] 202 Nozzle

[0037] A. First surface

[0038] B Second surface

[0039] M cells DETAILED DESCRIPTION

[0040] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0041] Please refer to 1 to Figure 2 It should be noted that the illustrations provided in this embodiment are only used to illustrate the basic concept of the present invention in a schematic manner, and the drawings only show components related to the present invention rather than being drawn according to the number, shape and size of components in actual implementation. In actual implementation, the type, quantity and proportion of each component may be changed arbitrarily, and the component layout may also be more complicated.

[0042] The present invention provides a cell analyzer, please refer to Figure 1 , which is a schematic diagram of the cross-sectional structure of the cell analyzer in one embodiment, includes a chip layer 100 and a flow channel layer 200 .

[0043] Specifically, the chip layer 100 includes a first surface A and a second surface B that are arranged opposite to each other. A liquid inlet 101, an impedance measurement unit 102 and a liquid driving unit 103 are provided in the chip layer 100. The liquid inlet 101 connects the first surface A and the second surface B. The impedance measurement unit 102 includes an electrode sheet 102a exposed on the first surface A. The liquid driving unit 103 includes a driving sheet 103a exposed on the first surface A.

[0044] Specifically, the flow channel layer 200 is combined with the first surface A of the chip layer 100, and the flow channel layer 200 includes a flow channel 201 and a nozzle 202. The flow channel 201 is opened on the side of the flow channel layer 200 facing the chip layer 100, the liquid inlet 101, the flow channel 201 and the nozzle 202 are connected in sequence, and the outlet of the nozzle 202 is opened on the side of the flow channel layer 200 away from the chip layer 100, at least a part of the electrode sheet 102a is located in the flow channel 201, and at least a part of the driving sheet 103a faces the outlet of the nozzle 202.

[0045] Specifically, the liquid driving unit 103 drives the liquid to be tested to be ejected from the nozzle 202 through the driving sheet 103 a exposed on the first surface A of the chip layer 100 , and drives the liquid to be tested to flow in the flow channel 201 .

[0046] As an example, the chip layer 100 includes a silicon substrate or other suitable semiconductor substrate, and the liquid driving unit 103 also includes a driving circuit integrated in the chip layer 100 and electrically connected to the driving chip 103a. When the liquid to be tested is added to the liquid inlet 101 and naturally flows into the flow channel 201 and the nozzle 202, the spraying of the nozzle 202 is controlled by an electrical signal to complete the precise quantitative transportation of the liquid without the need for an external liquid driving device, which can greatly reduce the volume of the cell analysis instrument.

[0047] Specifically, the total volume of the liquid to be tested sprayed can be calculated according to the single spray volume and the number of sprays of a single nozzle.

[0048] In some embodiments, the liquid driving unit 103 drives the liquid by thermal bubble driving. The driving plate 103a includes a heating film. The heating film is heated by the electrical signal of the driving circuit. The instantaneous high temperature of the heating film vaporizes the liquid above, generating bubbles to drive the liquid to flow and spray out from the nozzle 202. Then, the subsequent liquid is replenished under the action of capillary force, thereby providing power for the continuous flow of the liquid.

[0049] In another embodiment, the liquid driving unit 103 may also drive the liquid by piezoelectric drive. The driving plate 103a includes a piezoelectric crystal plate. The electrical signal of the driving circuit causes the electric driving plate 103a to deform, and the liquid to be tested is squeezed and sprayed out from the nozzle 202. The subsequent liquid is then replenished under the action of capillary force, thereby providing power for the continuous flow of the liquid.

[0050] It should be noted that when the liquid driving unit 103 drives the liquid in a piezoelectric manner, the chip layer 100 is provided with a back cavity (not shown) at a corresponding position of the piezoelectric crystal sheet to provide a deformation space.

[0051] Specifically, the impedance measurement unit 102 is used to count specific cells in the liquid to be tested. The measurement principle is that when the liquid to be tested flows in the flow channel 201, if a cell M passes through the impedance measurement unit 102, it will change the voltage difference between the electrode sheets 102a, thereby generating an impedance signal, wherein different cells such as red blood cells and white blood cells generate different impedance signals when passing through the electrode sheets of the impedance measurement unit 102, so that specific cells can be counted according to the number of times a specific impedance signal appears.

[0052] In some embodiments, the impedance measurement unit 102 includes two electrode sheets 102a.

[0053] In other embodiments, the impedance measurement unit 102 includes more than two electrode sheets 102a, for example, 4, 6 or 8 electrode sheets 102a, wherein a greater number of electrodes leads to a more accurate impedance measurement result, but a larger area may be occupied, which can be weighed based on actual needs.

[0054] Specifically, the concentration of the specific cells in the liquid to be tested can be further calculated based on the count of the specific cells and the calculated total volume of the liquid to be tested that is sprayed.

[0055] In some embodiments, in order to increase the flux, the number of the flow channels 201 is multiple, the number of the nozzles 202 is multiple, and each of the flow channels 201 is connected to a corresponding nozzle 202 .

[0056] As an example, see Figure 2 , which is a plan view of the cell analyzer in one embodiment, includes two nozzles 202 , two flow channels 201 and two impedance measurement units 102 .

[0057] In some embodiments, the number of the nozzles 202 is preferably greater than 100, wherein the more the flow channels 201 and the more the nozzles 202 are, the higher the throughput of the cell analyzer and the faster the analysis speed.

[0058] Specifically, the nozzle 202 and the impedance measurement unit 102 constitute a drive detection unit. In some embodiments, the drive detection unit is relatively small, for example, the width is less than 50 microns, so that hundreds or thousands of drive detection units can be integrated on a single chip, which can achieve rapid counting of cells in the liquid to be tested.

[0059] As an example, micromachining technology is used to complete micron-scale flow channel processing, so as to achieve single-cell counting of the liquid to be tested and increase the accuracy of counting.

[0060] In some embodiments, at least two of the flow channels 201 have different passing sizes to achieve separation of cells of different particle sizes, that is, large and small cells can enter different flow channels respectively. In specific applications, the flow channels can be designed with corresponding sizes for cells of specific sizes.

[0061] The cell analyzer of the present invention can be manufactured using semiconductor micro-electromechanical technology. For example, a silicon substrate or other suitable semiconductor substrate is first provided, and the required circuit structure is manufactured using a CMOS process. Then, the driving plate of the liquid driving unit 103 and the electrode plate of the impedance measurement unit 102 are processed on the substrate using a micro-electromechanical (MEMS) process, and then the flow channel layer 200 is manufactured.

[0062] In some embodiments, the flow channel layer 200 includes a photolithographic polymer material, such as photosensitive polyimide, photosensitive epoxy resin, etc.

[0063] In some embodiments, the flow channel layer 200 is made of SU8 material, which is a polymer material whose main components are epoxy resin and photosensitizer. It has the advantages of high resolution, high precision, high stability, etc., and can prepare micron-level structures.

[0064] In some embodiments, the manufacturing of the flow channel layer 200 includes the following steps:

[0065] (1) bonding a first photolithographic polymer material layer 200a on the chip layer 100, and patterning the first photolithographic polymer material layer 200a by a photolithography process to obtain the flow channel 201, wherein the bonding may be to adhere the first photolithographic polymer material layer 200a to the chip layer 100, and the first photolithographic polymer material layer 200a is tightly adhered to the surface of the chip layer 100 under the action of atmospheric pressure;

[0066] (2) Bonding a second photolithographic polymer material layer 200b onto the first photolithographic polymer material layer 200a, and patterning the second photolithographic polymer material layer 200b using a photolithography process to obtain the nozzle 202, wherein the bonding can be to adhere the second photolithographic polymer material layer 200b to the first photolithographic polymer material layer 200a, and the second photolithographic polymer material layer 200b is tightly adhered to the surface of the first photolithographic polymer material layer 200a under the action of atmospheric pressure.

[0067] The present invention also provides a cell analysis method, comprising the following steps:

[0068] The liquid to be tested is added into the liquid inlet, and a driving plate facing the nozzle outlet is used to drive the liquid to be tested to be ejected from the nozzle and drive the liquid to be tested to flow in the flow channel;

[0069] Using an impedance measurement unit to measure the impedance of cells flowing through the impedance measurement unit in the flow channel and calculate the number of specific cells;

[0070] Calculate the total volume of the liquid to be tested sprayed based on the single spray volume and the number of sprays of a single nozzle;

[0071] The concentration of the specific cells in the liquid to be tested is calculated based on the calculated number of specific cells and the total volume of the liquid to be tested that is ejected.

[0072] In one application scenario, the liquid to be tested is blood. In other application scenarios, the liquid to be tested may also be other liquids containing cells, or other liquids containing biological particles.

[0073] The following uses blood particle analysis as an example to illustrate the cell analysis process:

[0074] (1) Blood containing various particles is added into the chip layer 100 through the liquid inlet 101, and the blood naturally fills the entire flow channel until the nozzle 202. Due to the surface tension of the liquid, the liquid stops flowing after filling the entire flow channel;

[0075] (2) Start the thermal bubble drive nozzle to stimulate the liquid above the thermal bubble drive sheet to spray to the outside of the chip, driving the blood at the liquid inlet to continuously flow through the electrode sheet. At the same time, the electrode impedance is detected. If particles such as red blood cells and white blood cells pass through the electrode, different impedance signals will be generated. The blood particles are analyzed by counting different cells, and the total volume of the sprayed blood is calculated based on the single nozzle single spray volume and the number of sprays, and then the blood particle concentration is calculated.

[0076] The cell analyzer of the present invention utilizes inkjet-driven liquid technology to realize miniaturization, automation and low cost of whole blood analysis or other cell analysis equipment, thereby making the analysis equipment a home health care detection device, realizing home-based disease pre-detection, and avoiding high hospital load and cross infection during pandemics. The cell analyzer of the present invention can be manufactured based on CMOS-MEMS inkjet technology, with integrated impedance electrodes to realize on-chip measurement of liquid particles to be tested, high-throughput electrodes and inkjet nozzles integrated at the chip level to realize rapid analysis of the liquid to be tested, and accurate quantification of the ejection volume of a single nozzle of an inkjet printing nozzle to realize precise control of the analysis volume of the liquid to be tested.

[0077] In summary, the cell analyzer of the present invention includes a chip layer and a flow channel layer, wherein the chip layer includes a first surface and a second surface arranged oppositely, a liquid inlet, an impedance measurement unit and a liquid drive unit are provided in the chip layer, the liquid inlet communicates with the first surface and the second surface of the chip layer, the impedance measurement unit includes an electrode sheet exposed on the first surface of the chip layer, the liquid drive unit includes a drive sheet exposed on the first surface of the chip layer, the flow channel layer is combined with the first surface of the chip layer and includes a flow channel and a nozzle, the flow channel is opened on the side of the flow channel layer facing the chip layer, the liquid inlet, the flow channel and the nozzle are connected in sequence, the outlet of the nozzle is opened on the side of the flow channel layer away from the chip layer, at least a part of the electrode sheet is located in the flow channel, and at least a part of the drive sheet faces the outlet of the nozzle. The cell analyzer of the present invention can complete the precise quantitative transportation of the liquid to be tested and the accurate counting of specific cells, realize the precise measurement of the number and concentration of cells on the chip, and has the characteristics of miniaturization, automation and low cost. Therefore, the present invention effectively overcomes various shortcomings in the prior art and has a high industrial utilization value.

[0078] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A cell analyzer, characterized in that: include: A chip layer, comprising a first surface and a second surface arranged opposite to each other, wherein a liquid inlet, an impedance measuring unit and a liquid driving unit are provided in the chip layer, wherein the liquid inlet communicates with the first surface and the second surface, the impedance measuring unit comprises an electrode sheet exposed on the first surface, and the liquid driving unit comprises a driving sheet exposed on the first surface; A flow channel layer is combined with the first surface of the chip layer, the flow channel layer includes a flow channel and a nozzle, the flow channel is opened on the side of the flow channel layer facing the chip layer, the liquid inlet, the flow channel and the nozzle are connected in sequence, the outlet of the nozzle is opened on the side of the flow channel layer away from the chip layer, at least a part of the electrode sheet is located in the flow channel, and at least a part of the drive sheet faces the outlet of the nozzle.

2. The cell analyzer according to claim 1, characterized in that: The liquid driving unit drives the liquid in a manner including thermal bubble driving or piezoelectric driving.

3. The cell analyzer according to claim 1, characterized in that: There are multiple flow channels, multiple nozzles, and each flow channel is connected to a corresponding nozzle.

4. The cell analyzer according to claim 3, characterized in that: At least two of the flow channels have different passage sizes to achieve separation of cells with different particle sizes.

5. The cell analyzer according to claim 1, characterized in that: The impedance measurement unit includes at least two electrode sheets.

6. The cell analyzer according to claim 1, characterized in that: The liquid driving unit also includes a driving circuit integrated in the chip layer and electrically connected to the driving chip.

7. The cell analyzer according to claim 1, characterized in that: The number of the nozzles is greater than 100.

8. The cell analyzer according to claim 1, characterized in that: The liquid to be tested includes blood.

9. The cell analyzer according to claim 1, characterized in that: The flow channel layer includes a photolithographic polymer material.

10. A cell analysis method, characterized in that: The following steps are involved: The liquid to be tested is added into the liquid inlet, and a driving plate facing the nozzle outlet is used to drive the liquid to be tested to be ejected from the nozzle and drive the liquid to be tested to flow in the flow channel; Using an impedance measurement unit to measure the impedance of cells flowing through the impedance measurement unit in the flow channel and calculate the number of specific cells; Calculate the total volume of the liquid to be tested sprayed based on the single spray volume and the number of sprays of a single nozzle; The concentration of the specific cells in the liquid to be tested is calculated based on the calculated number of specific cells and the total volume of the liquid to be tested that is ejected.

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