Cytometer and cell detection method

Abstract

The invention provides a cytometer and a cell rotation method, the cytometer comprises a piezoelectric substrate made of a piezoelectric material and a transducer, and the piezoelectric substrate comprises a sample placing area for placing cell sap to be detected; the transducer comprises a first interdigital transducer and a second interdigital transducer which are oppositely arranged on the piezoelectric substrate, and the sample placement area is located between the first interdigital transducer and the second interdigital transducer; wherein the first interdigital transducer and the second interdigital transducer are used for driving the piezoelectric substrate to vibrate so as to spread surface acoustic waves in cell sap in the sample placement area, the width of an area corresponding to the cell sap in the sample placement area is related to the wavelength of the surface acoustic waves, and the surface acoustic waves are used for forming at least two vortexes in the cell sap; the vortex drives cells in the cell sap to rotate orderly, the area width is the width of the cell sap in the propagation direction of the surface acoustic wave, the cells in multiple vortexes can be clearly observed in the cell rotation process, and in-situ control over multiple biological particles is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of cell detection, and in particular to a cytometer and a cell detection method. Background Art

[0002] In the field of cell morphology analysis, cell pathology analysis is currently mainly performed through microscopic examination of cell samples. Traditional cytopathology examination methods based on two-dimensional image analysis cannot detect the cell characteristics in three-dimensional space, resulting in false negative results in cell detection and reducing the sensitivity of screening.

[0003] Currently, cell-based three-dimensional inspection primarily relies on optical tweezers and electric field methods to control cell rotation and manipulation, but this can damage cells or require direct contact with them. While cell inspection methods using acoustofluidics devices can manipulate cell populations without direct contact, existing acoustofluidics technology can typically only achieve in situ manipulation of single biological particles, resulting in low in situ detection throughput. Summary of the Invention

[0004] The present application provides a cytometer and a cell detection method, which aims to use an acoustic fluidic device for cell inspection. Although the method can realize the manipulation of cell groups without direct contact, the existing acoustic fluidic technology can usually only realize the in situ manipulation of single biological particles, resulting in the problem of low in situ detection throughput.

[0005] In a first aspect, the present application provides a cytometer, comprising:

[0006] A piezoelectric substrate, wherein the piezoelectric substrate is made of a piezoelectric material and comprises a sample placement area for placing a cell fluid to be detected;

[0007] a transducer, the transducer comprising a first interdigital transducer and a second interdigital transducer, the first interdigital transducer and the second interdigital transducer being arranged opposite to each other on the piezoelectric substrate, and the sample placement area being located between the first interdigital transducer and the second interdigital transducer;

[0008] In which, the first interdigital transducer and the second interdigital transducer are used to drive the piezoelectric substrate to vibrate so as to propagate surface acoustic waves in the cell fluid in the sample placement area. The width of the area corresponding to the cell fluid in the sample placement area is related to the wavelength of the surface acoustic wave. The surface acoustic wave is used to form at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner. The width of the area is the width of the cell fluid along the propagation direction of the surface acoustic wave.

[0009] In a second aspect, the present application provides a cell detection method, which is applied to a cytometer, wherein the cytometer includes a piezoelectric substrate and a transducer, wherein the piezoelectric substrate is made of a piezoelectric material, and the piezoelectric substrate includes a sample placement area, wherein the sample placement area is used to place the cell fluid to be detected; the transducer includes a first interdigital transducer and a second interdigital transducer, wherein the first interdigital transducer and the second interdigital transducer are arranged relative to each other on the piezoelectric substrate, and the sample placement area is located between the first interdigital transducer and the second interdigital transducer; the method includes:

[0010] Determining the emission parameters of the first and second IDTs according to the width of the area corresponding to the cell fluid in the sample placement area, wherein the area width is the width of the cell fluid along the propagation direction of the surface acoustic wave;

[0011] According to the emission parameters, the first and second interdigital transducers are controlled to drive the piezoelectric substrate to vibrate to propagate surface acoustic waves in the cell fluid. The wavelength of the surface acoustic waves is related to the width of the region. The surface acoustic waves drive the piezoelectric substrate to vibrate and thereby form at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner.

[0012] The present application provides a cytometer. By configuring the width of the area corresponding to the cell fluid in the sample placement area of ​​the cytometer and the surface acoustic wave generated by the piezoelectric substrate driven by the first and second interdigital transducers, the surface acoustic wave simultaneously forms at least two nodes in the cell fluid, allowing the cells to be captured and fixed in two vortices. Ultimately, the cells rotate in an orderly manner under the drive of the vortices. The provided cytometer can clearly observe cells within multiple vortices during cell rotation, achieving in situ manipulation of multiple biological particles, thereby improving both the throughput and accuracy of cell detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following is a brief introduction to the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0014] Figure 1 is a schematic diagram of the structure of the cytometer provided in an embodiment of the present application;

[0015] Figure 2 This is a schematic diagram of the principle of cell rotation provided in an embodiment of the present application;

[0016] Figure 3 is a structural diagram of a transducer provided in an embodiment of the present application;

[0017] Figure 4 is a schematic structural diagram of another cytometer provided in an embodiment of the present application;

[0018] Figure 5 This is a schematic diagram of a cell detection process provided in an embodiment of the present application;

[0019] Figure 6 is a schematic structural diagram of another cytometer provided in an embodiment of the present application;

[0020] Figure 7 This is a schematic diagram of the results of cell detection provided in the examples of the present application;

[0021] Figure 8 This is a schematic flow chart of the steps of a cell detection method provided by the present application;

[0022] Figure 9 This is a schematic diagram of an acoustic field of cell rotation provided in an embodiment of the present application;

[0023] Figure 10 Schematic diagram of the controllable cell rotation method provided in the examples of the present application;

[0024] Figure 11 This is a quantitative evaluation graph of cell morphology parameters observed from multiple angles, as provided in the examples of the present application.

[0025] Description of main components and symbols:

[0026] 10. Cytometer; 11. Piezoelectric substrate; 111. Sample placement area; 12. Transducer; 121. First IDT; 1211. First printed circuit board; 1212. First IDT unit; 122. Second IDT; 1221. Second printed circuit board; 1222. Second IDT unit; 13. Third printed circuit board; 131. First RF interface; 132. First probe; 14. Fourth printed circuit board; 141. Second RF interface; 142. Second probe; 15. Bottom plate; 151. Through hole; 16. Optical device; 17. Cell observation chamber; 18. Fixing device; 181. Pressing plate; 1811. Pipe through hole; 182. First bracket; 1821. First main bracket; 1822. First sub-bracket; 183. Second bracket; 1831. Second main bracket; 1832. Second sub-bracket; 184. Fixing plate; 185. Handle. DETAILED DESCRIPTION

[0027] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0028] The flowcharts shown in the accompanying drawings are for illustrative purposes only and do not necessarily include all contents and operations / steps, nor must they be executed in the order described. For example, some operations / steps may be decomposed, combined, or partially merged, so the actual execution order may vary depending on the actual situation.

[0029] It should be understood that the terms used in this specification are only for the purpose of describing specific embodiments and are not intended to limit the present application. As used in this specification and the appended claims, the singular forms "a", "an", and "the" are intended to include the plural forms unless the context clearly indicates otherwise.

[0030] It should be understood that, in order to facilitate a clear description of the technical solutions of the embodiments of the present application, in the embodiments of the present application, words such as "first" and "second" are used to distinguish between identical or similar items with substantially identical functions and effects. For example, the first interdigital transducer and the second interdigital transducer are merely used to distinguish between different transducers and do not limit their order. Those skilled in the art will understand that words such as "first" and "second" do not limit the quantity and execution order, and words such as "first" and "second" do not necessarily limit them to be different.

[0031] It should be further understood that the term “and / or” used in this specification and the appended claims refers to any and all possible combinations of one or more of the associated listed items, and includes these combinations.

[0032] To facilitate understanding of the embodiments of the present application, some of the terms involved in the embodiments of the present application are briefly explained below.

[0033] 1. Piezoelectric material: Piezoelectric material is a functional material characterized by direct piezoelectric effect and inverse piezoelectric effect. Under the action of mechanical force, the centers of positive and negative charges in the piezoelectric material will shift, resulting in polarization; under the action of an electric field, the piezoelectric material will deform. Based on this principle, when a regular voltage is applied, the piezoelectric material will produce a regular deformation movement, thereby generating vibration. In this process, the acoustic waves generated by the piezoelectric material include surface acoustic waves (SAW) and bulk acoustic waves (BAW), but for the vibration of the material, surface acoustic waves play a key role.

[0034] 2. Transducer: A device that converts one form of energy, such as electrical, mechanical, or acoustic energy, into another is called a transducer, also known as an active sensor. The transducer is the core component of ultrasonic equipment, and its characteristic parameters determine the performance of the entire device.

[0035] A transducer is an energy conversion device, and its performance description and evaluation require many parameters. Transducer characteristic parameters include resonant frequency, bandwidth, electromechanical coupling coefficient, electroacoustic efficiency, mechanical quality factor, impedance characteristics, frequency characteristics, directivity, and transmit and receive sensitivity. Different transducers have different performance requirements. During the specific transducer design process, the relevant parameters must be rationally designed based on the specific application.

[0036] 3. Interdigital Transducer (IDT): An IDT is a metal pattern shaped like the crossed fingers of two hands formed on the surface of a piezoelectric substrate. Its function is to achieve acoustic-to-electrical transduction. IDTs are mainly used in pairs in surface acoustic wave devices. When an alternating electrical signal is applied to the input end of a set of IDTs on the piezoelectric substrate, a periodically distributed electric field is generated. Due to the inverse piezoelectric effect, corresponding elastic deformation is stimulated near the surface of the piezoelectric medium, causing the vibration of solid particles and forming surface acoustic waves that propagate along the surface of the substrate. When the surface acoustic wave reaches the other end of the piezoelectric medium, the positive piezoelectric effect generates charges at both ends of the metal electrodes, allowing another set of IDTs to output an alternating electrical signal.

[0037] 4. Standing waves: Two waves propagate in opposite directions but vibrate in the same direction, with the same amplitude and frequency. On the waveform, the positions of the nodes and antinodes of a standing wave remain constant, giving the impression of being "stationary," but their instantaneous values ​​change over time. If the amplitudes of the two waves are equal, the amplitude of the nodes is zero. Every point on the combined wave undergoes simple harmonic oscillations of the same period.

[0038] Since the horizontal distance between two adjacent wave nodes is still half a wavelength, the wavefront of a standing wave consists of a series of antinodes and wave nodes. Although the height of the wavefront at the antinodes varies periodically, the horizontal position of this section is fixed, so the position of the wave nodes of the standing wave is also fixed.

[0039] 5. Acoustic tweezers technology: Acoustic tweezers technology, also known as acoustofluidics, is an all-round manipulation tool in the field of life sciences. It can perform operations such as displacement, rotation, separation, enrichment, washing, sorting, lysis, fusion, and phenotypic detection on exosomes, viruses, cells, sperm, bacteria, microorganisms, parasites, nematodes, zebrafish, organoids, and droplets.

[0040] Acoustic tweezers, leveraging the principles of ultrasound and fluid dynamics, is an innovative crossover technology based on a microfluidic platform. It offers the advantages of non-contact, label-free, and non-invasive manipulation of bioparticles, enabling efficient, precise, and cost-effective manipulation of bioparticles. Furthermore, the corresponding acoustic tweezers platform and tools can be rapidly designed based on the specific application scenario of bioparticle manipulation.

[0041] As an ideal biomedical application tool, acoustic tweezers technology is widely used and has great potential in specific scenarios such as CTC (Circulating Tumor Cell) sorting, cell washing, flow cytometry, exosome enrichment and separation, and iPSC (Induced pluripotent stem cells) directed differentiation.

[0042] The following describes some embodiments of the present application in detail with reference to the accompanying drawings. In the absence of conflict, the following embodiments and features therein may be combined with each other.

[0043] Please refer to Figure 1 , Figure 1 This is a schematic diagram of the structure of the cytometer provided in the embodiment of the present application. Figure 1 As shown, the provided cytometer 10 includes a piezoelectric substrate 11 and a transducer 12. The piezoelectric substrate 11 is made of piezoelectric material and includes a sample placement area 111 for placing the cell fluid to be detected. The transducer 12 includes a first interdigital transducer 121 and a second interdigital transducer 122. The first interdigital transducer 121 and the second interdigital transducer 122 are arranged relative to each other on the piezoelectric substrate 11, and the sample placement area 111 is located between the first interdigital transducer 121 and the second interdigital transducer 122.

[0044] Among them, the first interdigital transducer 121 and the second interdigital transducer 122 are used to drive the piezoelectric substrate 11 to vibrate so as to propagate surface acoustic waves in the cell fluid in the sample placement area 111. The width of the area corresponding to the cell fluid in the sample placement area is related to the wavelength of the surface acoustic wave. The surface acoustic wave is used to form at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner. The area width is the width of the cell fluid along the propagation direction of the surface acoustic wave.

[0045] Specifically, since the provided cytometer 10 includes a piezoelectric substrate 11 made of a piezoelectric material and a first interdigital transducer 121 and a second interdigital transducer 122 arranged relatively on the piezoelectric substrate 11, the first interdigital transducer 121 and the second interdigital transducer 122 simultaneously transmit electrical signals toward the sample placement area 111 of the piezoelectric substrate 11, thereby being able to drive the piezoelectric substrate 11 to vibrate through the inverse piezoelectric effect of the piezoelectric material, thereby propagating the cell fluid in the sample placement area 111. Figure 1 The two surface acoustic waves shown have opposite vibration propagation directions, the same amplitude and the same frequency. The standing wave formed by the superposition of the two surface acoustic waves with opposite propagation directions as traveling waves can form at least two vortices in the cell fluid.

[0046] Please also refer to Figure 1 and Figure 2 , Figure 2 This is a schematic diagram of the principle of cell rotation provided in the embodiment of the present application. Figure 1 for Figure 2 Schematic diagram of the cross section in the AA′ direction. Figure 1 and Figure 2 As shown, the cytometer 10 provided herein designs the width of the area corresponding to the cell fluid in the sample placement area 111 according to the wavelength of the surface acoustic wave (SAW), so that the SAW has at least two nodes in the cell fluid, forming at least two vortexes. Consequently, cells in the cell fluid can be captured and fixed in the vortexes after entering the sample placement area 111, forming a linear arrangement and rotating in the direction of the vortexes.

[0047] It should be noted that if Figure 2 As shown, the cells in the cell fluid rotate in situ with the y-axis as the axis in the vortex. At the same time, when the cell fluid 10 completes the detection of the cells, new cell fluid to be detected can be injected into the sample placement area 111. At this time, the cell fluid that has completed the detection will flow out of the sample placement area 111.

[0048] In some embodiments, the provided piezoelectric substrate 11 is made of any one of a lithium niobate substrate, a lead zinc niobate-lead titanate substrate, a lead magnesium zirconate titanate substrate, a lead zinc niobate titanate substrate, a lead nickel niobate titanate substrate, or a polyvinylidene fluoride substrate. The piezoelectric properties of the piezoelectric material can enable the piezoelectric substrate 11 to cooperate with the first interdigital transducer 121 and the second interdigital transducer 122 to realize the transmission of surface acoustic waves.

[0049] Exemplarily, the provided piezoelectric substrate 11 is made of a piezoelectric material of a lithium niobate substrate. Since the lithium niobate substrate has excellent piezoelectric properties, for example, a 128-degree Y-cut lithium niobate substrate can cooperate well with the first interdigital transducer 121 and the second interdigital transducer 122 to realize the transmission of surface acoustic waves. At the same time, the lithium niobate substrate has good light transmittance, which can also facilitate the provided cytometer 10 to observe the state of cells rotating in the vortex.

[0050] It should be noted that the width of the area corresponding to the cell fluid in the sample placement area is related to the wavelength of the surface acoustic wave. The correlation can be specifically that the area width is n times the wavelength of the surface acoustic wave, where n is an integer greater than or equal to 1, so that the surface acoustic wave forms at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner. The area width is the width of the cell fluid along the propagation direction of the surface acoustic wave. The two vortices can provide real-time reference during cell detection, thereby improving the accuracy of cell inspection and at the same time improving the efficiency of cell detection.

[0051] It should also be noted that the region width is n times the wavelength of the surface acoustic wave, and n may also be a decimal greater than 1, such as 1.5, 1.6, etc.

[0052] Of course, it is understandable that in order for the surface acoustic wave to form at least two vortices in the cell fluid, and the vortices drive the cells in the cell fluid to rotate in an orderly manner, the area width can be equal to or slightly smaller than the wavelength of the surface acoustic wave. The "slightly smaller" is, for example, an area width equal to 0.99λ-0.98λ, where λ is the wavelength of the surface acoustic wave. That is, the area width that can generate two vortices is within the scope of protection of this application, regardless of whether it is the best effect.

[0053] The width of the area corresponding to the presence of cell fluid in the sample placement area is related to the wavelength of the surface acoustic wave. It can also be understood that the difference between the area width and n times the wavelength of the surface acoustic wave is within a preset difference range. For example, when n is equal to 1.5, three vortices can be formed in the cell fluid, when n is equal to 2, four vortices can be formed in the cell fluid, and when n is equal to 2.5, five vortices can be formed in the cell fluid. From a mathematical formula, the relationship between the area width and the wavelength of the surface acoustic wave is as follows:

[0054] Δ=0.5×k×λ-a

[0055] Where Δ represents a preset difference, for example, a value between -10 μm and 10 μm, k represents the number of eddies, λ represents the wavelength of the surface acoustic wave, and a represents the area width. The cytometer 10 provided in this embodiment of the present application can adjust the wavelength and area width of the surface acoustic wave according to the desired number of eddies, thereby achieving multi-channel cell detection.

[0056] It can also be understood that the ratio of the area width to n times the wavelength of the surface acoustic wave is within a preset magnification range. For example, when n is equal to 1.5, 2, and 2.5 in the above analysis, three vortices, four vortices, and five vortices can be formed in the cell, respectively. The relationship between the area width and the wavelength of the surface acoustic wave is as follows:

[0057]

[0058] Where p represents a preset ratio, for example, a value selected from a preset magnification range of 0.95 to 1.05, k represents the number of eddies, λ represents the wavelength of the surface acoustic wave, and a represents the area width. The cytometer 10 provided in the embodiment of the present application can adjust the wavelength and area width of the surface acoustic wave according to the desired number of eddies, thereby achieving multi-channel cell detection.

[0059] In some embodiments, the difference between the area width and one wavelength of the surface acoustic wave is within a preset difference range, so that two vortices are formed in the cell fluid. Since the number of vortices formed in the cell fluid is related to the number of nodes of the surface acoustic wave in the sample placement area 111, by controlling the difference between the area width and the wavelength of the surface acoustic wave to be within the preset difference range, it is ensured that the two nodes in the surface acoustic wave are located in the sample placement area, thereby forming two vortices in the cell fluid. When using the cytometer 10 provided in the embodiment of the present application for cell detection, it is possible to increase the throughput of cell detection while also conducting experiments such as dual-channel comparative detection to improve the accuracy of cell detection.

[0060] For example, the preset difference range can be -10 μm to 10 μm. When the area width is greater than or less than the wavelength of the surface acoustic wave within 10 μm, the surface acoustic wave can have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortices in the cell fluid of the sample placement area 111.

[0061] It should be noted that the preset difference range is related to the unit of wavelength. In some embodiments, for example, the preset difference range can be -6μm to 6μm. When the area width is greater than or less than the wavelength of the surface acoustic wave within 6μm, it can further ensure that the surface acoustic wave can have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortices in the cell fluid of the sample placement area 111.

[0062] For example, the area width of the cytometer 10 may range from 195 μm to 205 μm, and the wavelength of the surface acoustic wave emitted by the first IDT 121 and the second IDT 122 may range from 194.8 μm to 204.4 μm. The area width and surface acoustic wave wavelength selected within the aforementioned area width range and emitted surface acoustic wave wavelength range ensure that two nodes of the surface acoustic wave are located in the sample placement area 111 of the cytometer 10, thereby forming two vortexes in the cell fluid in the sample placement area 111.

[0063] In some embodiments, the ratio of the area width to the wavelength of the surface acoustic wave is within a preset magnification range so that two vortices are formed in the cell fluid. Since the number of vortices formed in the cell fluid is related to the number of nodes of the surface acoustic wave in the sample placement area 111, by controlling the ratio of the area width to the wavelength of the surface acoustic wave within a preset magnification range, it is ensured that the two nodes in the surface acoustic wave are located in the sample placement area, thereby forming two vortices in the cell fluid. When using the cytometer 10 provided in the embodiment of the present application for cell detection, it is possible to increase the throughput of cell detection while also conducting experiments such as dual-channel comparative detection to improve the accuracy of cell detection.

[0064] Exemplarily, the preset magnification range is 0.95 to 1.05. When the ratio of the area width being greater than or less than the surface acoustic wave does not exceed 5% of the wavelength of the surface acoustic wave, the surface acoustic wave can have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortices in the cell fluid of the sample placement area 111.

[0065] It should be noted that, in some embodiments, the preset magnification range is 0.97 to 1.03. When the ratio of the area width being greater than or less than the surface acoustic wave does not exceed 3% of the wavelength of the surface acoustic wave, it can further ensure that the surface acoustic wave can have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortices in the cell fluid of the sample placement area 111, and the double helix has the best detection effect.

[0066] Exemplarily, the waveform of the surface acoustic wave is a sine wave. By sending a sinusoidal excitation signal to the first interdigital transducer 121 and the second interdigital transducer 122, the first interdigital transducer 121 and the second interdigital transducer 122 drive the piezoelectric substrate 11 to vibrate so that the surface acoustic wave propagating in the cell fluid in the sample placement area 111 is a sine wave.

[0067] Exemplarily, the excitation radio frequency range of the surface acoustic wave is 19.41 MHz to 19.81 MHz, thereby forming two vortexes in the cell fluid. By controlling the excitation radio frequency range of the surface acoustic wave, for example, by setting the excitation radio frequency range for the surface acoustic wave emitted by the first IDT 121 and the second IDT 122, the difference / ratio between the wavelength and the area width of the ultimately generated surface acoustic wave can be made to conform to a preset difference range / preset magnification range, thereby forming two vortexes in the cell fluid of the sample placement area 111.

[0068] Exemplarily, the peak-to-peak voltage of the surface acoustic wave is 7.5 volts to 17.5 volts, thereby forming two vortexes in the cell fluid. By controlling the driving voltage of the surface acoustic wave, for example, by setting a magnification range between the driving voltage and the preset peak voltage of the surface acoustic wave in the first and second interdigital transducers 121 and 122, the difference / ratio between the wavelength and the area width of the ultimately generated surface acoustic wave can conform to the preset difference range / preset magnification range, thereby forming two vortexes in the cell fluid of the sample placement area 111.

[0069] For example, please refer to Figure 3 , Figure 3 This is a schematic diagram of the structure of a transducer provided in an embodiment of the present application. Figure 3 As shown, the first IDT 121 and the second IDT 122 are composed of two rows of bus bars (bus bars), each row of bus bars has a plurality of electrodes, and these electrodes are spaced apart from each other to form an IDT.

[0070] It should be noted that, in some embodiments, the finger width of the first IDT 121 and the second IDT 122 ranges from 35.6 μm to 41.8 μm. The finger width of the IDT refers to the width of its electrodes, such as Figure 3 As shown by x in FIG. By limiting the finger width range of the first IDT 121 and the second IDT 122, it is further ensured that the surface acoustic waves generated by the piezoelectric material driven by the first IDT 121 and the second IDT 122 can have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortexes in the cell fluid in the sample placement area 111.

[0071] It should be noted that, in some embodiments, the finger spacing range of the first interdigital transducer 121 and the second interdigital transducer 122 is 58.1 μm to 64.1 μm. The finger spacing of the interdigital transducer refers to the distance between two adjacent electrodes above and below. By limiting the finger spacing range of the first interdigital transducer 121 and the second interdigital transducer 122, it can be further ensured that the surface acoustic wave generated by the piezoelectric material driven by the first interdigital transducer 121 and the second interdigital transducer 122 can have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortexes in the cell fluid of the sample placement area 111.

[0072] It should be noted that, in some embodiments, the aperture of the first IDT 121 and the second IDT 122 ranges from 5 mm to 15 mm. The aperture of the IDT refers to the acoustic aperture through which the IDT transmits signals, such as Figure 3 W in the figure, where 10 mm is taken as the optimal embodiment of the present application. By limiting the apertures of the first IDT 121 and the second IDT 122, it is further ensured that the surface acoustic waves generated by the piezoelectric material driven by the first IDT 121 and the second IDT 122 can have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortexes in the cell fluid of the sample placement area 111.

[0073] It should be noted that, in some embodiments, the number of electrode pairs of the first IDT 121 and the second IDT 122 is 40. Figure 3 A partial schematic diagram of the electrodes of the IDT in FIG. 1 shows that the first IDT 121 and the second IDT 122 include three electrode pairs. By limiting the number of electrode pairs of the first IDT 121 and the second IDT 122, it is further ensured that the surface acoustic waves generated by the piezoelectric material driven by the first IDT 121 and the second IDT 122 have two nodes located in the sample placement area 111 of the cytometer 10, thereby forming two vortexes in the cell fluid in the sample placement area 111.

[0074] In some embodiments, please refer to Figure 3 The first IDT 121 includes a first printed circuit board 1211 and a first interdigital unit 1212 disposed on a first side surface of the first printed circuit board 1211. The second IDT 122 includes a second printed circuit board 1221 and a second interdigital unit 1222 disposed on a first side surface of the second printed circuit board 1221. The first printed circuit board 1211 and the second printed circuit board 1221 are both disposed on the piezoelectric substrate 11, and the first sides of the first printed circuit board 1211 and the second printed circuit board 1221 are in contact with the piezoelectric substrate 11.

[0075] The cytometer 10 provided in the present application adds a first IDT 121 and a second IDT 122 to a printed circuit board, and then after the first IDT 121 and the second IDT 122 are aligned, they can jointly drive the piezoelectric substrate 11 to generate surface acoustic waves to form two vortices in the cell fluid.

[0076] It should be noted that, in some embodiments, Figure 4 As shown, Figure 4 This is a schematic diagram of the structure of another cytometer provided in an embodiment of the present application. A first printed circuit board 1211 and a second printed circuit board 1221 are integrally formed. The integrally formed circuit board body includes a hollow portion located between the first interdigital unit 1212 and the second interdigital unit 1222. The hollow portion corresponds to the sample placement area. This eliminates the need for alignment of the first and second interdigital transducers during each use, improving the efficiency of the provided cytometer 10.

[0077] It should be noted that, in some embodiments, the first IDT 121 includes a first interdigital unit 1212 and a metal layer on the surface of the first interdigital unit 1212. The second IDT 122 includes a second interdigital unit 1222 and a metal layer on the surface of the second interdigital unit 1222. The first interdigital unit 1212 and the second interdigital unit 1222 are both in contact with the piezoelectric substrate 11.

[0078] In some embodiments, the cytometer 10 further includes a signal generator, which is electrically connected to the first IDT 121 and the second IDT 122, respectively, and is used to control the first IDT 121 and the second IDT 122 to drive the piezoelectric substrate 11 to vibrate so as to propagate surface acoustic waves toward the cell fluid in the sample placement area 111. The signal generator sends an excitation electrical signal to the first IDT 121 and the second IDT 122, thereby controlling the first IDT 121 and the second IDT 122 to drive the piezoelectric substrate 11 to vibrate so as to propagate surface acoustic waves toward the cell fluid in the sample placement area 111, thereby forming at least two vortexes in the cell fluid.

[0079] Exemplarily, the signal generator can be any one of a magnetoelectric signal generator, a Hall signal generator and a photoelectric signal generator. By adjusting the parameters of the excitation electrical signal emitted by the signal generator, the first interdigital transducer 121 and the second interdigital transducer 122 can be controlled to drive the piezoelectric substrate 11 to vibrate so as to propagate surface acoustic waves to the cell fluid in the sample placement area 111. Therefore, the embodiment of the present application does not limit the type of signal generator.

[0080] For example, please refer to Figure 4The provided cytometer also includes a third printed circuit board 13 and a fourth printed circuit board 14. The third printed circuit board 13 is provided with a first radio frequency interface 131 (SMA interface) and a first probe 132. The first radio frequency connection port 131 is connected to the signal generator for receiving a first driving signal sent by the signal generator. The first probe 132 is connected to the first interdigital transducer 121 for inputting the first driving signal to the first interdigital transducer 121 to control the first interdigital transducer 121 to drive the piezoelectric substrate 11 to vibrate so as to transmit surface acoustic waves to the cell fluid in the sample placement area 111. A second RF interface 141 and a second probe 142 are provided on the fourth printed circuit board 14. The second RF connection port 141 is connected to the signal generator for receiving a second driving signal sent by the signal generator. The second probe 142 is connected to the second IDT 122 for inputting the second driving signal to the second IDT 122 to control the second IDT 122 to drive the piezoelectric substrate 11 to vibrate so as to propagate surface acoustic waves to the cell fluid in the sample placement area 111.

[0081] Through the arrangement of the third printed circuit board 13 and the fourth printed circuit board 14, the cytometer 10 provided in the present application can adjust the driving signals of the first interdigital transducer 121 and the second interdigital transducer 122 in real time through the signal generator, thereby ensuring that two eddy currents are generated in the cytometer.

[0082] It should be noted that, in some embodiments, Figure 4 As shown, there are through holes on both sides of the bottom plate 15 of the cytometer 10 for allowing the first probe 132 and the second probe 142 to be connected to the first IDT 121 and the second IDT 122 respectively, thereby realizing the integration of the cytometer 10.

[0083] In some embodiments, please refer to Figure 4 The provided cytometer 10 may further include a base plate 15 and an optical device 16. The piezoelectric substrate 11 is disposed on the base plate 15, which has a through hole 151 therein corresponding to the cell fluid. The optical device 16 is disposed within the through hole of the base plate 15, and is configured to accurately observe cell parameters of the cells during rotation at predetermined intervals through the through hole of the base plate 15, as the vortex drives the cells in the cell fluid to rotate in an orderly manner.

[0084] Please refer to Figure 5 , Figure 5 This is a schematic diagram of a cell detection process provided in the embodiment of the present application. Figure 5As shown, the provided cytometer 10 can cause cells in the cell fluid to rotate orderly through two vortexes from a chaotic state. During the cell rotation process, an optical device 16 is provided below the piezoelectric substrate 11 to observe the specific parameters of each cell in the two vortexes, thereby enabling accurate cell screening of each cell.

[0085] For example, the preset interval angle ranges from 5° to 60°. The preset interval angle for cell observation can be set according to the speed of the eddy current to adjust the interval time of the optical image of the cell taken by the optical device 16. The size of the preset interval angle is set according to the accuracy required for cell screening, for example Figure 4 By using 45 degrees as the preset interval angle, the parameters of each cell can be observed in all aspects, so that the provided cytometer 10 has more precise accuracy in cell detection.

[0086] Exemplarily, the cell parameters include at least one or more of nuclear solidity, nuclear circularity, and nuclear-to-cytoplasmic ratio (N / C). By using the cytometer 10 provided in the present application, key parameters for cell detection, such as nuclear solidity, nuclear circularity, and nuclear-to-cytoplasmic ratio, can be obtained through the optical device 16, thereby improving the cell detection effect of the cytometer while also reducing the manufacturing cost of the cytometer.

[0087] Exemplarily, the optical device 16 includes a polarizer, which ensures that only light in a vertical direction can be reflected into a cell detection device, such as a microscope, so that the optical image of the cells obtained will not cause ghosting, thereby improving the accuracy of cell detection.

[0088] Exemplarily, the base plate 15 can be made of any metal material such as copper, aluminum or stainless steel. The base plate 15 can not only support and stabilize the piezoelectric substrate 11 of the cytometer 10, but also dissipate heat generated by the vibration of the piezoelectric substrate 11 driven by the inverse piezoelectric effect.

[0089] In some embodiments, please also refer to Figure 1 and Figure 4 The provided cytometer 10 further includes a cell observation chamber 17, which is used to be placed in the sample placement area 111. The cell observation chamber 17 includes a cell fluid cavity and an inlet and an outlet connected to the cell fluid cavity (please refer to Figure 2 The direction from the inlet to the outlet is roughly parallel to the direction of the vortex. After the cell fluid is pumped into the cell fluid cavity of the cell observation chamber 17 from the inlet via a cell pump, the cells in the cell fluid are able to move in an orderly manner through the vortex within the sample placement area 111 and finally escape through the outlet of the cell observation chamber 17. The cytometer provided by the design of the cell observation chamber 17 can provide a carrier for cell detection.

[0090] Exemplarily, the direction from the inlet to the outlet of the cell observation chamber 17 is roughly parallel to the direction of the vortex, such as the angle difference between the inlet of the cell observation chamber and the direction of the vortex is within ±10°, thereby ensuring that the cells in the cell fluid flowing into the cell observation chamber can rotate in an orderly manner in the direction of the vortex.

[0091] Exemplarily, the side wall thickness of the shell of the cell observation chamber ranges from 2 mm to 8 mm, such as 5 mm, thereby ensuring that the generated surface acoustic wave can be accurately transmitted into the cell observation chamber to form at least two vortices.

[0092] For example, the cell fluid cavity is in the shape of a rectangular parallelepiped, and the width of the cell fluid cavity ranges from 195 μm to 205 μm. When the cell fluid cavity is in the shape of a rectangular parallelepiped (please refer to Figure 2 ), where the width of the cell fluid cavity is consistent with the width of the sample placement area 111. By setting the cell fluid cavity width to 195 μm to 205 μm and adjusting the wavelength of the surface acoustic wave, at least two vortices can be formed within the sample placement area 111.

[0093] Exemplarily, the length of the cell fluid cavity ranges from 10 mm to 20 mm, such as 15 mm. Sufficient length of the cell fluid cavity ensures that the provided cytometer 10 has enough time to observe cells during cell detection, thereby ensuring the accuracy of cell detection.

[0094] Exemplarily, the height of the cell fluid cavity ranges from 50 μm to 70 μm, such as 60 μm. The height of the cell fluid cavity is related to the amplitude of the surface acoustic wave. Setting the height of the cell fluid cavity can ensure that the cells have sufficient rotation space in the vortex.

[0095] Exemplarily, the shell material of the cell observation chamber 17 is polydimethylsiloxane. By using polydimethylsiloxane to make the shell of the cell observation chamber 17 , it can be ensured that the cell fluid in the cell observation chamber can stably move in the sample placement area 111 .

[0096] For example, please refer to Figure 6 , Figure 6 This is a schematic diagram of the structure of another cytometer provided in the embodiment of the present application. Figure 6 As shown, the cytometer 10 further includes a fixing device 18, which is used to fix the cell observation chamber 17 to the sample placement area 111. To improve the stability of the provided cytometer 10 during use, the setting of the fixing device 18 can prevent the cell observation chamber 17 from shaking due to slight collisions during use of the cytometer 10, ensuring that the cells in the cell observation chamber can stably rotate in the vortex.

[0097] For example, please refer to Figure 6 The fixing device 18 includes a pressure plate 181, which is connected to the bottom plate 15 of the cytometer 10 and is used to fix the cell observation chamber 17 to the sample placement area 111. The pressure plate 181 is provided with a pipe through-hole 1811, which is used to pass a cell fluid inlet pipe and a cell fluid outflow pipe. The cell fluid inlet pipe and the cell fluid outflow pipe are respectively connected to the inlet and outlet of the cell fluid cavity. This further improves the stability of the cell observation chamber 17 while also preventing the cell pump from affecting the stability of the cell observation chamber 17 when injecting cell fluid into the cell observation chamber 17.

[0098] For example, the provided cytometer 10 can be detachably connected between the pressure plate 181 and the base plate 15 of the cytometer 10 by providing threaded holes. The pressure plate 181 and the base plate 15 can also be connected by adsorption to a wall. The embodiment of the present application does not limit the connection method between the pressure plate 181 and the base plate 15.

[0099] It should be noted that, in some embodiments, the fixing device 18 further includes a fixing plate 184 , which is disposed between the pressing plate 181 and the cavity of the cell observation chamber 17 , thereby stabilizing the connection between the pressing plate 181 and the cell observation chamber 17 .

[0100] It should be noted that, in some embodiments, the fixing plate 184 is made of acrylic, which can reduce the development cost of the cytometer 10 while improving the stability of the cytometer 10 .

[0101] It should be noted that, in some embodiments, the fixing device 18 further includes: a first bracket 182 and a second bracket 183. The first bracket 182 includes a first main bracket 1821 and a first sub-bracket 1822. The first sub-bracket 1822 is mounted on the first main bracket 1821. The first main bracket 1821 and the first sub-bracket 1822 are used to cooperate with each other to support the side wall of the cell observation chamber 17. The first main bracket 1821 is connected to the bottom plate 15 of the cytometer 10. The second bracket 183 is arranged relative to the first bracket 182. The second bracket 183 includes a second main bracket 1831 and a second sub-bracket 1832. The second sub-bracket 1832 is mounted on the second main bracket 1831. The second main bracket 1831 and the second sub-bracket 1832 are used to cooperate with each other to support the side wall of the cell observation chamber 17. The second main bracket 1831 is connected to the bottom plate 15 of the cytometer 10. By disposing the first bracket 182 and the second bracket 183 , the side surface of the cell observation chamber 17 can be firmly stabilized in the sample placement area 111 , thereby comprehensively improving the stability of the cytometer 10 .

[0102] It should be noted that, in some embodiments, the fixing device 18 further includes a handle 185, which is connected to the first main support 1821 of the first support 182 and the second main support 1831 of the second support 183. Thus, the cytometer can be moved safely and stably via the handle 185.

[0103] The present invention provides a cytometer that, by adjusting the width of the cell fluid region within the sample placement area and the surface acoustic wave (SAW) generated by a piezoelectric substrate driven by a first interdigital transducer and a second interdigital transducer, simultaneously generates at least two nodes in the cell fluid, trapping cells in two vortexes. Ultimately, the cells rotate in an orderly manner driven by the vortexes, thereby improving cell detection throughput.

[0104] Please refer to Figure 4 , Figure 4 FIG1 is a schematic diagram of the structure of another cytometer provided in an embodiment of the present application. The cytometer 10 provided includes a base plate 15, an optical device 16, a piezoelectric substrate 11, a transducer 12, and a cell observation chamber 17. A through hole 151 is defined in the base plate 15, and the optical device 16 is disposed within the through hole 151 of the base plate 15. The piezoelectric substrate 11 is disposed on the base plate 15 and is made of a piezoelectric material. The piezoelectric substrate 15 includes a sample placement area 111. The transducer 12 includes a first interdigital transducer 121 and a second interdigital transducer 122. The first interdigital transducer 121 and the second interdigital transducer are relatively arranged on the piezoelectric substrate 11, and the sample placement area 111 is located between the first interdigital transducer 121 and the second interdigital transducer 122. The cell observation chamber 17 is used to be placed in the sample placement area 111. The cell observation chamber 17 includes a cell fluid cavity and an inlet and an outlet connected to the cell fluid cavity. The cell fluid to be detected flows into the cell fluid cavity through the inlet and flows out of the cell fluid cavity through the outlet.

[0105] The first interdigital transducer 121 and the second interdigital transducer 122 are used to drive the piezoelectric substrate 11 to vibrate so as to propagate surface acoustic waves in the cell fluid in the sample placement area 111. The width of the region corresponding to the presence of the cell fluid in the sample placement area 111 is related to the wavelength of the surface acoustic wave. The surface acoustic wave is used to form at least two vortices in the cell fluid, and the vortices drive the cells in the cell fluid to rotate in an orderly manner. The region width is the width of the cell fluid along the propagation direction of the surface acoustic wave, and in the process of the vortex driving the cells in the cell fluid to rotate in an orderly manner, the cell parameters of the cells during the rotation process are observed at a preset interval angle through the optical device 16.

[0106] Specifically, since the provided cytometer 10 includes a piezoelectric substrate 11 made of a piezoelectric material and a first interdigital transducer 121 and a second interdigital transducer 122 arranged relatively on the piezoelectric substrate 11, the first interdigital transducer 121 and the second interdigital transducer 122 simultaneously transmit electrical signals toward the sample placement area 111 of the piezoelectric substrate 11, thereby being able to drive the piezoelectric substrate 11 to vibrate through the inverse piezoelectric effect of the piezoelectric material, thereby propagating the cell fluid in the sample placement area. Figure 1 The two acoustic waves shown have opposite vibration directions, the same amplitude, and the same frequency. The standing wave formed by the superposition of two surface acoustic waves with opposite propagation directions as traveling waves can form at least two eddies in the cell fluid. At the same time, by setting the optical device 16 in the through hole 151 of the bottom plate 15, the provided cytometer 10 can make the cells in the cell fluid rotate in an orderly manner through the two eddies. Figure 7 As shown, Figure 7 This is a schematic diagram of the results of cell detection provided in the examples of this application. Figure 7 As shown, compared with the traditional method of using pathological slides for microscopic examination, the provided cytometer 10 is clearer and more obvious in the detection of various cell parameters. Therefore, it can be proved that the provided cytometer can clearly observe the specific parameters of each cell in the two vortexes during the cell rotation, and then can perform accurate cell screening on each cell.

[0107] The relationship between the region width and the wavelength of the surface acoustic wave, as well as the definition of the parameters of the surface acoustic wave, can be referred to the above embodiments and will not be elaborated here.

[0108] For example, the preset interval angle ranges from 5° to 60°. The preset interval angle for cell observation can be set according to the speed of the eddy current to adjust the interval time of the optical image of the cell taken by the optical device 16. The size of the preset interval angle is set according to the accuracy required for cell screening, for example Figure 4 By using 45 degrees as the preset interval angle, the parameters of each cell can be observed in all aspects, so that the provided cytometer 10 has more precise accuracy in cell detection.

[0109] Exemplarily, the cell parameters include at least one or more of nuclear density, nuclear roundness and nuclear-cytoplasmic ratio. By using the cytometer 10 provided in the present application, key parameters for cell detection, such as nuclear density, nuclear roundness and nuclear-cytoplasmic ratio, can be obtained through the optical device 16, thereby improving the cell detection effect of the cytometer while also reducing the manufacturing cost of the cytometer.

[0110] Exemplarily, the optical device 16 includes a polarizer, which ensures that only light in a vertical direction can be reflected into a cell detection device, such as a microscope, so that the optical image of the cells obtained will not cause ghosting, thereby improving the accuracy of cell detection.

[0111] The present application provides a cytometer. By setting the width of the area corresponding to the cell fluid in the sample placement area of ​​the cytometer and the surface acoustic wave formed by the piezoelectric substrate driven by the first interdigital transducer and the second interdigital transducer, the surface acoustic wave simultaneously forms at least two wave nodes in the cell fluid, so that the cells are captured and fixed in two vortexes. Ultimately, the cells rotate in an orderly manner under the drive of the vortexes, and the parameters of the cell rotation process can be clearly observed through optical devices. The provided cytometer can clearly observe cells in multiple vortexes during the cell rotation process, realize in situ manipulation of multiple biological particles, thereby improving the cell detection throughput and the accuracy of cell detection.

[0112] Please refer to Figure 8 , Figure 8 The present invention provides a flowchart illustrating the steps of a cell detection method. The method is applied to the cytometer provided in any embodiment of the present invention. The cytometer includes a piezoelectric substrate and a transducer. The piezoelectric substrate is made of a piezoelectric material and includes a sample placement area for placing the cell fluid to be detected. The transducer includes a first interdigital transducer and a second interdigital transducer, which are arranged relative to each other on the piezoelectric substrate, and the sample placement area is located between the first interdigital transducer and the second interdigital transducer.

[0113] like Figure 8 As shown, the provided method includes steps S101 to S102.

[0114] S101. Determine emission parameters of the first IDT and the second IDT according to the width of the area corresponding to the cell fluid in the sample placement area, where the area width is the width of the cell fluid along the propagation direction of the surface acoustic wave.

[0115] Specifically, the cytometer can determine the parameters of the surface acoustic wave propagating in the sample placement area by obtaining the width of the cell fluid in the sample placement area. Based on the surface acoustic wave parameters, the transmission parameters of the first and second interdigital transducers can be determined. This ensures that the generated surface acoustic wave has the same number of nodes located in the sample placement area as the number of eddy currents to be generated.

[0116] S102. Control the first interdigital transducer and the second interdigital transducer to drive the piezoelectric substrate to vibrate according to the emission parameters to propagate surface acoustic waves in the cell fluid. The wavelength of the surface acoustic wave is related to the width of the area. The surface acoustic wave drives the piezoelectric substrate to vibrate and then forms at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner.

[0117] Specifically, the cytometer can control the first and second interdigital transducers to drive the piezoelectric substrate to vibrate through the inverse piezoelectric effect to propagate surface acoustic waves in the cell fluid based on the determined emission parameters of the first and second interdigital transducers. Since the surface acoustic wave has at least two nodes in the sample placement area, at least two vortices can be formed in the cell fluid, allowing the cells in the cell fluid to rotate in an orderly manner in the vortices, and also to complete the detection of the cells by acquiring optical images of the rotating cells. The provided method can also improve the accuracy of cell detection while improving the cell detection throughput.

[0118] It should be noted that the width of the area corresponding to the cell fluid in the sample placement area is related to the wavelength of the surface acoustic wave. The correlation can be specifically that the area width is n times the wavelength of the surface acoustic wave, where n is an integer greater than or equal to 1, so that the surface acoustic wave forms at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner. The area width is the width of the cell fluid along the propagation direction of the surface acoustic wave. The two vortices can provide real-time reference during cell detection, thereby improving the accuracy of cell inspection and at the same time improving the efficiency of cell detection.

[0119] It should also be noted that the region width is n times the wavelength of the surface acoustic wave, and n may also be a decimal greater than 1, such as 1.5, 1.6, etc.

[0120] Of course, it is understandable that in order for the surface acoustic wave to form at least two vortices in the cell fluid, and the vortices drive the cells in the cell fluid to rotate in an orderly manner, the area width can be equal to or slightly smaller than the wavelength of the surface acoustic wave. The "slightly smaller" is, for example, an area width equal to 0.99λ-0.98λ, where λ is the wavelength of the surface acoustic wave. That is, the area width that can generate two vortices is within the scope of protection of this application, regardless of whether it is the best effect.

[0121] The width of the area corresponding to the presence of cell fluid in the sample placement area is related to the wavelength of the surface acoustic wave. It can also be understood that the difference between the area width and n times the wavelength of the surface acoustic wave is within a preset difference range. For example, when n is equal to 1.5, three vortices can be formed in the cell fluid, when n is equal to 2, four vortices can be formed in the cell fluid, and when n is equal to 2.5, five vortices can be formed in the cell fluid. From a mathematical formula, the relationship between the area width and the wavelength of the surface acoustic wave is as follows:

[0122] Δ=0.5×k×λ-a

[0123] Where Δ represents a preset difference, for example, a value between -10 μm and 10 μm, k represents the number of eddies, λ represents the wavelength of the surface acoustic wave, and a represents the area width. The cytometer provided in the embodiments of the present application can adjust the wavelength and area width of the surface acoustic wave according to the desired number of eddies, thereby enabling multi-channel cell detection.

[0124] It can also be understood that the ratio of the area width to n times the wavelength of the surface acoustic wave is within a preset magnification range. For example, when n is equal to 1.5, 2, and 2.5 in the above analysis, three vortices, four vortices, and five vortices can be formed in the cell, respectively. The relationship between the area width and the wavelength of the surface acoustic wave is as follows:

[0125]

[0126] Where p represents a preset ratio, for example, a number selected from a preset magnification range of 0.95 to 1.05, k represents the number of eddies, λ represents the wavelength of the surface acoustic wave, and a represents the area width. The cytometer provided in the embodiments of the present application can adjust the wavelength and area width of the surface acoustic wave according to the desired number of eddies, thereby achieving multi-channel cell detection.

[0127] In some embodiments, the difference between the area width and one wavelength of the surface acoustic wave is within a preset difference range, so that two vortices are formed in the cell fluid. Since the number of vortices formed in the cell fluid is related to the number of nodes of the surface acoustic wave in the sample placement area, by controlling the difference between the area width and the wavelength of the surface acoustic wave to be within a preset difference range, it is ensured that the two nodes in the surface acoustic wave are located in the sample placement area, thereby forming two vortices in the cell fluid. When using the cytometer provided in the embodiment of the present application for cell detection, it is possible to increase the throughput of cell detection while also conducting experiments such as dual-channel comparative detection to improve the accuracy of cell detection.

[0128] For example, the preset difference range can be -10 μm to 10 μm. When the area width is greater than or less than the wavelength of the surface acoustic wave within 10 μm, the surface acoustic wave can have two nodes located in the sample placement area of ​​the cytometer, thereby forming two vortices in the cell fluid in the sample placement area.

[0129] It should be noted that the preset difference range is related to the unit of wavelength. In some embodiments, for example, the preset difference range can be -6μm to 6μm. When the area width is greater than or less than the wavelength of the surface acoustic wave within 6μm, it can further ensure that the surface acoustic wave can have two nodes located in the sample placement area of ​​the cytometer, thereby forming two vortices in the cell fluid in the sample placement area.

[0130] For example, the cytometer's zone width can range from 195 μm to 205 μm, and the wavelengths of the surface acoustic waves emitted by the first and second IDTs can range from 194.8 μm to 204.4 μm. Within the aforementioned zone width ranges and emitted surface acoustic wave wavelength ranges, the zone widths and surface acoustic wave wavelengths selected ensure that two nodes of the surface acoustic wave are located in the sample placement area of ​​the cytometer, thereby forming two vortices in the cytosol in the sample placement area.

[0131] In some embodiments, the ratio of the area width to the wavelength of the surface acoustic wave is within a preset magnification range so that two vortices are formed in the cell fluid. Since the number of vortices formed in the cell fluid is related to the number of nodes of the surface acoustic wave in the sample placement area, by controlling the ratio of the area width to the wavelength of the surface acoustic wave within a preset magnification range, it is ensured that the two nodes in the surface acoustic wave are located in the sample placement area, thereby forming two vortices in the cell fluid. When using the cytometer provided in the embodiment of the present application for cell detection, it is possible to increase the throughput of cell detection while also conducting experiments such as dual-channel comparative detection to improve the accuracy of cell detection.

[0132] Exemplarily, the preset magnification range is 0.95 to 1.05. When the ratio of the area width being greater than or less than the surface acoustic wave does not exceed 5% of the wavelength of the surface acoustic wave, the surface acoustic wave can have two nodes located in the sample placement area of ​​the cytometer, thereby forming two vortices in the cell fluid in the sample placement area.

[0133] It should be noted that, in some embodiments, the preset magnification range is 0.97 to 1.03. When the ratio of the area width being greater than or less than the surface acoustic wave does not exceed 3% of the wavelength of the surface acoustic wave, it can further ensure that the surface acoustic wave can have two nodes located in the sample placement area of ​​the cytometer, thereby forming two vortices in the cell fluid in the sample placement area, and the double helix has the best detection effect.

[0134] Exemplarily, the waveform of the surface acoustic wave is a sine wave, and by sending a sinusoidal excitation signal to the first interdigital transducer and the second interdigital transducer, the first interdigital transducer and the second interdigital transducer drive the piezoelectric substrate 11 to vibrate so that the surface acoustic wave propagating in the cell fluid in the sample placement area is a sine wave.

[0135] Exemplarily, the excitation radio frequency range of the surface acoustic wave is 19.41 MHz to 19.81 MHz, thereby forming two vortexes in the cell fluid. By controlling the excitation radio frequency range of the surface acoustic wave, for example, by setting the excitation radio frequency ranges for the surface acoustic wave emission of the first and second interdigital transducers, the difference / ratio between the wavelength and the region width of the ultimately generated surface acoustic wave can be adjusted to meet a preset difference range / preset magnification range, thereby forming two vortexes in the cell fluid of the sample placement area.

[0136] Exemplarily, the peak-to-peak voltage of the surface acoustic wave is 7.5 volts to 17.5 volts, thereby forming two cellular vortices in the cell fluid. By controlling the driving voltage of the surface acoustic wave, for example, by setting a magnification range between the driving voltage and the preset peak voltage of the surface acoustic wave in the first and second interdigital transducers, the difference / ratio between the wavelength and the area width of the ultimately generated surface acoustic wave can be adjusted to meet the preset difference range / preset magnification range, thereby forming two vortices in the cell fluid of the sample placement area.

[0137] It should be noted that, in some embodiments, please refer to Figure 9 , Figure 9 The method provided in this application numerically simulates the acoustic field of different area widths and cross sections of the sample placement area. Figure 9 (a) Shows the maximum sound pressure changing with the height and width of the cell observation chamber. Figure 9 The three insets in (b) show the acoustic pressure at three different cell observation chamber heights with the same region width of 200 μm, where the black arrows indicate the nodes. Figure 9 (c) Ⅰ represents the first-order sound pressure in the cell observation chamber with a size of 60 μm (height) and 200 μm (width). As can be seen in the figure, two wave nodes are formed in the sample observation chamber. Figure 9 II in (c) represents the time-averaged second-order velocity at the same size. As can be seen in the figure, two vortices are formed in the sample observation chamber, and the arrows in the figure indicate the directions of the vortices.

[0138] In some embodiments, please refer to Figure 4 The cytometer also includes a base plate and an optical device. The piezoelectric substrate is arranged on the base plate, and a through hole corresponding to the cell fluid is provided in the base plate; the optical device is arranged in the through hole of the base plate; the method also includes: in the process of eddy current driving the cells in the cell fluid to rotate in an orderly manner, obtaining an optical path image of the cells in the rotation process at a preset interval angle through the optical device; and obtaining cell parameters of the cells based on the optical path image.

[0139] Please refer to Figure 7 Compared with the traditional method of using pathological slides for microscopic examination, the provided method can detect various cell parameters more clearly and obviously. The provided method can clearly observe the specific parameters of each cell in the two vortexes during the cell rotation, and thus can perform accurate cell screening on each cell.

[0140] Exemplarily, the preset interval angle ranges from 5° to 60°. The preset interval angle for cell observation can be set according to the speed of the eddy current to adjust the interval time of the optical image of the cell taken by the optical device. The size of the preset interval angle is set according to the accuracy required for cell screening, for example Figure 4 By setting 45 degrees as the preset interval angle, the parameters of each cell can be observed in all aspects, making the provided cytometer more precise and accurate in cell detection.

[0141] It should be noted that, in some embodiments, please refer to Figure 10 , Figure 10 Schematic diagram of the controllable cell rotation method provided in the embodiment of the present application. Figure 10 As shown, the method provided in this application can accurately control the rotation of cells in two vortexes, and the parameters of the cell rotation process can also be clearly observed through the provided method.

[0142] Exemplarily, the cell parameters include at least one or more of nuclear density, nuclear roundness and nuclear-cytoplasmic ratio. By using the cytometer provided in the present application, key parameters for cell detection, such as nuclear density, nuclear roundness and nuclear-cytoplasmic ratio, can be obtained through optical devices, thereby improving the cell detection effect of the cytometer while also reducing the manufacturing cost of the cytometer.

[0143] It should be noted that, in some embodiments, please refer to Figure 11 , Figure 11 This is a quantitative evaluation graph of cell morphology parameters observed from multiple angles provided in the examples of this application. Figure 11 It can be seen that the nuclear density, nuclear roundness and nuclear-cytoplasmic ratio of 30 cells can be monitored at an interval of 45°, and thus the cells in the multi-vortex flow can be accurately and comprehensively evaluated.

[0144] Exemplarily, the optical device includes a polarizer, which ensures that only light in a vertical direction can be reflected into a cell detection device, such as a microscope, so that the optical image of the cells obtained will not cause ghosting, thereby improving the accuracy of cell detection.

[0145] The present application provides a cell detection method, which can clearly observe cells in multiple vortices during cell rotation, thereby improving the cell detection throughput and the accuracy of cell detection.

[0146] A computer-readable storage medium is also provided in an embodiment of the present application. The computer-readable storage medium stores a computer program. The computer program includes program instructions. The processor executes the program instructions to implement the steps of the cell detection method provided in the above embodiment.

[0147] The specific implementation of the above operations can be found in the previous embodiments and will not be repeated here.

[0148] The computer-readable storage medium may be an internal storage unit of the computer device in the aforementioned embodiment, such as a hard disk or memory of the computer device. The computer-readable storage medium may also be an external storage device of the computer device, such as a plug-in hard disk, a smart media card (SMC), a secure digital (SD) card, a flash memory card, etc. equipped on the computer device.

[0149] Since the computer program stored in the computer-readable storage medium can execute any of the cell detection methods provided in the embodiments of the present application, the beneficial effects that can be achieved by any of the cell detection methods provided in the embodiments of the present application can be achieved. Please refer to the previous embodiments for details and will not be repeated here.

[0150] The serial numbers of the embodiments of the present application are for description only and do not represent the advantages or disadvantages of the embodiments. The above are only specific implementation methods of the present application, but the scope of protection of the present application is not limited thereto. Any technician familiar with the technical field can easily think of various equivalent modifications or replacements within the technical scope disclosed in this application, and these modifications or replacements should be included in the scope of protection of this application. Therefore, the scope of protection of this application should be based on the scope of protection of the claims.

Claims

1. A cytometer, characterized in that The cytometer comprises: A piezoelectric substrate, wherein the piezoelectric substrate is made of a piezoelectric material and comprises a sample placement area for placing a cell fluid to be detected; a transducer, the transducer comprising a first interdigital transducer and a second interdigital transducer, the first interdigital transducer and the second interdigital transducer being arranged opposite to each other on the piezoelectric substrate, and the sample placement area being located between the first interdigital transducer and the second interdigital transducer; In which, the first interdigital transducer and the second interdigital transducer are used to drive the piezoelectric substrate to vibrate so as to propagate surface acoustic waves in the cell fluid in the sample placement area. The width of the area corresponding to the cell fluid in the sample placement area is related to the wavelength of the surface acoustic wave. The surface acoustic wave is used to form at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner. The width of the area is the width of the cell fluid along the propagation direction of the surface acoustic wave.

2. The cytometer according to claim 1, wherein The difference between the area width and the wavelength of the surface acoustic wave is within a preset difference range, or the ratio of the area width to the wavelength of the surface acoustic wave is within a preset ratio range, so that two vortices are formed in the cell fluid.

3. The cytometer according to claim 2, characterized in that The preset difference range is -10 μm to 10 μm; or, the preset magnification range is 0.95 to 1.

05.

4. The cytometer according to claim 2, wherein The width of the region ranges from 195 μm to 205 μm, and the wavelength of the surface acoustic waves emitted by the first IDT and the second IDT ranges from 194.8 μm to 204.4 μm.

5. The cytometer according to claim 2, characterized in that The excitation radio frequency range of the surface acoustic wave is 19.41 MHz to 19.81 MHz; or, the peak-to-peak voltage of the surface acoustic wave is 7.5 volts to 17.5 volts.

6. The cytometer according to claim 1, characterized in that The finger width range of the first interdigital transducer and the second interdigital transducer is 35.6μm to 41.8μm; or, the finger spacing range of the first interdigital transducer and the second interdigital transducer is 58.1μm to 64.1μm; or, the aperture of the first interdigital transducer and the second interdigital transducer is 10mm; or, the number of electrode pairs of the first interdigital transducer and the second interdigital transducer is 40.

7. The cytometer according to claim 1, wherein The first interdigital transducer comprises: a first printed circuit board and a first interdigital unit arranged on a first side surface of the first printed circuit board; the second interdigital transducer comprises: a second printed circuit board and a second interdigital unit arranged on a first side surface of the second printed circuit board; The first printed circuit board and the second printed circuit board are both disposed on the piezoelectric substrate, and the first side surfaces of the first printed circuit board and the second printed circuit board are in contact with the piezoelectric substrate.

8. The cytometer according to claim 7, characterized in that The first printed circuit board and the second printed circuit board are integrally formed, and the integrally formed circuit board body includes a hollow portion located between the first interdigital unit and the second interdigital unit, and the hollow portion corresponds to the sample placement area.

9. The cytometer according to claim 1, wherein The cytometer further comprises: A signal generator is electrically connected to the first IDT and the second IDT respectively, and is used to control the first IDT and the second IDT to drive the piezoelectric substrate to vibrate so as to propagate the surface acoustic wave to the cell fluid in the sample placement area.

10. The cytometer according to claim 9, characterized in that The cytometer further comprises: a third printed circuit board, on which a first radio frequency interface and a first probe are provided, wherein the first radio frequency interface is connected to the signal generator and is used to receive a first driving signal sent by the signal generator, and the first probe is connected to the first interdigital transducer and is used to input the first driving signal to the first interdigital transducer to control the first interdigital transducer to drive the piezoelectric substrate to vibrate so as to propagate the surface acoustic wave to the cell fluid in the sample placement area; A fourth printed circuit board is provided with a second radio frequency interface and a second probe, the second radio frequency connection port is connected to the signal generator for receiving a second drive signal sent by the signal generator, the second probe is connected to the second interdigital transducer for inputting the second drive signal to the second interdigital transducer to control the second interdigital transducer to drive the piezoelectric substrate to vibrate so as to propagate the surface acoustic wave to the cell fluid in the sample placement area.

11. The cytometer according to claim 1, wherein The cytometer further comprises: A bottom plate, the piezoelectric substrate is arranged on the bottom plate, and the bottom plate is provided with a through hole corresponding to the cell fluid; An optical device is arranged in the through hole of the bottom plate, and is used to observe the cell parameters of the cells during the rotation process at preset interval angles through the optical device when the vortex drives the cells in the cell fluid to rotate in an orderly manner.

12. The cytometer according to claim 11, characterized in that The preset interval angle has an angle range of 5° to 60°; or, the cell parameters include at least one or more of nuclear density, nuclear roundness and nuclear-cytoplasmic ratio; or, the optical device includes a polarizer.

13. The cytometer according to claim 1, wherein The cytometer further comprises: A cell observation chamber is used to be placed in the sample placement area, and the cell observation chamber includes a cell fluid cavity and an inlet and an outlet connected to the cell fluid cavity; the direction from the inlet to the outlet is roughly parallel to the direction of the vortex.

14. The cytometer according to claim 13, wherein: The side wall thickness of the shell of the cell observation chamber ranges from 2 mm to 8 mm; or, the cell fluid cavity is in the shape of a rectangular parallelepiped, and the width of the cell fluid cavity ranges from 195 μm to 205 μm; or, the length of the cell fluid cavity ranges from 10 mm to 20 mm; or, the height of the cell fluid cavity ranges from 50 μm to 70 μm.

15. The cytometer according to claim 14, characterized in that The cytometer further comprises: A fixing device is used to fix the cell observation chamber on the sample placement area.

16. The cytometer according to claim 15, characterized in that The fixing device includes a pressing plate, which is connected to the bottom plate of the cytometer and is used to fix the cell observation chamber in the sample placement area. The pressing plate is provided with a pipeline through hole, and the pipeline through hole is used to pass a cell fluid inflow tube and a cell fluid outflow tube. The cell fluid inflow tube and the cell fluid outflow tube are respectively connected to the inlet and outlet of the cell fluid cavity.

17. The cytometer according to claim 16, wherein: The fixing device further comprises: a first bracket, the first bracket comprising a first main bracket and a first sub-bracket, the first sub-bracket being mounted on the first main bracket, the first main bracket and the first sub-bracket being used to cooperate in supporting a side wall of the cell observation chamber, and the first main bracket being connected to a bottom plate of the cytometer; The second bracket is arranged relative to the first bracket, the second bracket includes a second main bracket and a second sub-bracket, the second sub-bracket is installed on the second main bracket, the second main bracket and the second sub-bracket are used to cooperate to support the side wall of the cell observation chamber, and the second main bracket is connected to the bottom plate of the cytometer.

18. The cytometer according to claim 1, wherein The cytometer further comprises: A bottom plate, wherein a through hole is provided in the bottom plate, and the piezoelectric substrate is arranged on the bottom plate; an optical device, wherein the optical device is disposed in the through hole of the base plate; A cell observation chamber, the cell observation chamber is used to be placed in the sample placement area, the cell observation chamber includes a cell fluid cavity and an inlet and an outlet connected to the cell fluid cavity, the cell fluid to be detected flows into the cell fluid cavity through the inlet and flows out of the cell fluid cavity through the outlet; Wherein, in the process of the vortex driving the cells in the cell fluid to rotate in an orderly manner, the cell parameters of the cells in the rotation process are observed at preset interval angles through the optical device.

19. A cell detection method, characterized in that: The invention is applied to a cytometer, the cytometer comprising a piezoelectric substrate and a transducer, the piezoelectric substrate being made of a piezoelectric material and comprising a sample placement area for placing a cell fluid to be detected; the transducer comprising a first interdigital transducer and a second interdigital transducer, the first interdigital transducer and the second interdigital transducer being arranged oppositely on the piezoelectric substrate, the sample placement area being located between the first interdigital transducer and the second interdigital transducer; the method comprising: Determining the emission parameters of the first and second IDTs according to the width of the area corresponding to the cell fluid in the sample placement area, wherein the area width is the width of the cell fluid along the propagation direction of the surface acoustic wave; According to the emission parameters, the first and second interdigital transducers are controlled to drive the piezoelectric substrate to vibrate to propagate surface acoustic waves in the cell fluid. The wavelength of the surface acoustic waves is related to the width of the region. The surface acoustic waves drive the piezoelectric substrate to vibrate and thereby form at least two vortices in the cell fluid. The vortices drive the cells in the cell fluid to rotate in an orderly manner.

20. The method according to claim 19, characterized in that The cytometer further includes a base plate and an optical device, the piezoelectric substrate is disposed on the base plate, and a through hole corresponding to the cell fluid is provided in the base plate; the optical device is disposed in the through hole of the base plate; and the method further includes: In the process of the eddy current driving the cells in the cell fluid to rotate in an orderly manner, obtaining optical path images of the cells in the rotation process at preset interval angles through the optical device; The cell parameters of the cell are obtained according to the optical path image; wherein the angle range of the preset interval angle is 5° to 60°; or, the cell parameters include at least one or more of nuclear density, nuclear roundness and nuclear-cytoplasmic ratio; or, the optical device includes a polarizer.

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