Scanning flow cytometer

Abstract

The present invention discloses a scanning flow cytometer, comprising: a liquid flow focusing module, an optical signal excitation detection module, a scanning galvanometer, an acquisition and control board, and a host computer. The present invention focuses the laser into a light spot smaller than the size of a cell, and realizes cell imaging by light spot scanning, which can improve the power density of the imaging light source and effectively excite fluorescence; when the scanning excitation method is adopted in the present invention, the entire laser spot is used for imaging, which has an overall excitation effect and does not have the situation of uneven distribution; at the same time, it can overcome the problem of uneven excitation of the light spot power caused by unstable liquid flow, which leads to a decrease in imaging quality. In the present invention, by setting a detection pinhole on the focal plane of the objective lens, the stray light outside the focal plane can be blocked, and the spherical aberration and chromatic aberration can be eliminated; by using a photomultiplier tube to collect the light signal, very weak signals can be amplified, and the imaging sensitivity can be greatly improved compared with the traditional CCD.

Description

Technical Field

[0001] The invention relates to the technical field of biological particle detection, and in particular to a scanning flow cell imaging analyzer. Background Art

[0002] Flow cytometry is a technique that can rapidly and quantitatively analyze and sort cells or other biological particles arranged in a single file in a liquid flow. When it is necessary to obtain information on cell morphology and internal structure, imaging flow cytometers have greater advantages than traditional flow cytometers. It combines the high throughput of the flow system with the subcellular resolution of the imaging system, which not only makes up for the deficiency of traditional flow cytometers in being unable to distinguish subcellular structural features and morphological information, but also improves the disadvantage of the optical microscopic imaging system that is not suitable for large-scale cell observation due to the low imaging rate. The fine morphological information of cells such as size and shape, spectral information such as probe type and position, and the characteristics of high-throughput detection make it important in biological research such as analyzing cell types, cell cycles, and intracellular signal transduction.

[0003] At present, Amnis, a subsidiary of Merck Millipore in the United States, has produced imaging flow cytometers with good performance, including Image StreamMark II and Flow Sight, which can capture up to 12 high-resolution images of each flow cell in real time, and the detection rate can reach 5000 cells / second. However, the image of each point on the field light source target will be disturbed by the diffraction or scattered light of the adjacent points, resulting in a decrease in imaging quality. In addition, the uneven Gaussian distribution of the laser spot and the disturbance of the liquid flow system will further cause a decrease in imaging accuracy. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a scanning flow cytometer for imaging analysis in view of the deficiencies in the above-mentioned prior art.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is: a scanning flow cytometer, comprising:

[0006] A liquid flow focusing module, which provides a sample flow channel and enables particles or cells to flow in a single file in the sample flow channel through focusing;

[0007] An optical signal excitation detection module, which is used to generate a laser spot and focus it on the detection area of ​​the sample flow channel to excite the particles or cells flowing through the detection area to generate fluorescence and scattered light, and to collect the fluorescence and scattered light signals generated by the particles or cells, and convert the optical signal into an electrical signal;

[0008] A scanning galvanometer, which is used to make the laser spot generated by the detection module excited by the optical signal to perform reciprocating scanning along the orthogonal direction of the particle or cell streamline in the detection area;

[0009] An acquisition and control board, which receives the electrical signal transmitted by the optical signal excitation detection module;

[0010] And a host computer, which receives the signal transmitted by the acquisition and control board and performs imaging.

[0011] Preferably, the laser spot generated by the light signal excitation detection module is smaller than the size of the particle or cell to be detected, so as to satisfy the point-by-point imaging of different regions of the particle or cell.

[0012] Preferably, the scanning galvanometer scans the laser spot back and forth perpendicular to the flow direction of the particles or cells, and the scanning period of the scanning galvanometer is no greater than the ratio of the length of the spot on the particle / cell streamline to the flow velocity of the particles / cells.

[0013] Preferably, the optical signal excitation detection module comprises a main optical path unit, at least one group of laser emission optical paths and at least one group of optical signal detection optical paths corresponding to the laser emission optical paths;

[0014] The laser emission optical path generates laser and transmits it to the main optical path unit. The main optical path unit forms a laser spot and focuses it on the particles or cells in the detection area. The main optical path unit also collects the fluorescence and scattered light generated by the particles or cells in the detection area and transmits it to the optical signal detection optical path. The optical signal detection optical path collects fluorescence and scattered light signals and converts them into electrical signals and then transmits them to the acquisition and control board.

[0015] Preferably, the main optical path unit comprises a first cylindrical lens, a second cylindrical lens, an excitation pinhole, a first dichroic mirror and an objective lens which are sequentially arranged along the incident direction of the laser, and a detection pinhole and a collecting mirror which are sequentially arranged along the optical path direction of the fluorescence and scattered light emitted by the objective lens;

[0016] The laser emission optical path includes a laser;

[0017] The optical signal detection optical path includes an interference filter, a lens and a photomultiplier tube. The light entering the optical signal detection optical path passes through the interference filter and the lens in sequence and then reaches the photomultiplier tube. The photomultiplier tube converts the optical signal into an electrical signal and transmits it to the acquisition and control board.

[0018] Preferably, the laser emitted by the laser emission optical path passes through the first cylindrical lens, the second cylindrical lens, and the excitation pinhole in sequence to form a laser spot, and then is reflected to the scanning galvanometer by the first dichroic mirror, and then is focused on the particles or cells in the detection area by the objective lens;

[0019] The fluorescence and scattered light generated by the particles or cells in the detection area are collected by the objective lens, pass through the first dichroic mirror, and then pass through the detection pinhole and the collecting mirror in sequence, and then enter the optical signal detection optical path.

[0020] Preferably, the number of the laser emission optical paths is N, and N is greater than 1, and the laser emission optical path further comprises a reflector or a dichroic mirror, so that the lasers emitted by the N lasers are all guided into the first cylindrical mirror through the reflector or the dichroic mirror;

[0021] The number of the optical signal detection optical paths is M, and the optical signal detection optical paths further include a dichroic mirror, so as to guide the light of M different wavelength ranges emitted by the collecting mirror into M different photomultiplier tubes through the dichroic mirror.

[0022] Preferably, the scanning galvanometer includes a bracket with an opening in the middle, a reflective plate suspended in the opening of the bracket by two symmetrically distributed elastic arms, and a piezoelectric plate arranged on the side arm of the bracket, and the piezoelectric plate is used to generate an inverse piezoelectric effect under the action of an electric field, thereby driving the reflective plate to twist.

[0023] Preferably, the liquid flow focusing module comprises a sample flow channel and a sheath liquid flow channel arranged at the periphery of the sample flow channel, and the sheath liquid in the sheath liquid flow channel causes the particles or cells in the sample flow channel to present a single-file flow state.

[0024] Preferably, the liquid flow focusing module includes a sample flow channel, a first piezoelectric element arranged on the sample flow channel for achieving focusing of particles or cells within the plane of the sample flow channel, and a second piezoelectric element arranged on the sample flow channel for achieving focusing of particles or cells in the depth direction of the sample flow channel.

[0025] The beneficial effects of the present invention are:

[0026] 1. The present invention focuses the laser into a light spot smaller than the size of the cell, and uses light spot scanning to achieve cell imaging, which can increase the power density of the imaging light source and effectively stimulate fluorescence;

[0027] 2. Traditional imaging uses field light sources, and the image of each point on the target will be disturbed by the diffraction or scattered light of adjacent points. The present invention uses point scanning technology to decompose the target into countless points in two-dimensional space, and uses a very fine laser beam to scan the image point by point and line by line, and then combines it into an overall plane image through a computer, so that the clarity and precision of the image are improved (the field light source used in traditional imaging illuminates a field, and all points on the cell are illuminated and imaged at the same time. The present invention uses point irradiation, and only a local point is illuminated each time during the scanning process, and other parts are not illuminated, so there is no interference from other parts, thereby overcoming the defect that traditional imaging is easily disturbed by the diffraction or scattered light of adjacent points).

[0028] 3. After shaping, the intensity of the laser spot is generally Gaussian distributed. If there is an uneven distribution of the spot intensity in the area where the cells pass, the imaging quality will be reduced. When the scanning excitation method is adopted in the present invention, the entire laser spot is used for imaging, which has an overall excitation effect and does not have the problem of uneven distribution. At the same time, it can overcome the problem of uneven excitation of the spot power caused by unstable liquid flow, which leads to reduced imaging quality.

[0029] 4. In the present invention, by arranging a detection pinhole on the focal plane of the objective lens, stray light outside the focal plane can be blocked, thereby eliminating spherical aberration and chromatic aberration.

[0030] 5. The present invention uses a photomultiplier tube to collect light signals, which can amplify very weak signals, and the imaging sensitivity is greatly improved compared to traditional CCDs.

[0031] 6. In some embodiments of the present invention, sound waves are used to focus cells, so that the flow rate is slowed down, which is beneficial to improving the subsequent detection sensitivity and imaging quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 is a schematic structural diagram of a scanning flow cytometric imaging analyzer in an embodiment of the present invention;

[0033] Figure 2 It is a schematic diagram of the principle of scanning particles or cells by a laser spot in an embodiment of the present invention;

[0034] Figure 3 is a schematic structural diagram of a scanning flow cytometric imaging analyzer in Example 1 of the present invention;

[0035] Figure 4 Schematic diagram of the structure of the laser emission optical path in Example 2 of the present invention;

[0036] Figure 5 It is a schematic diagram of the structure of the optical signal detection optical path in Embodiment 2 of the present invention;

[0037] Figure 6 is a schematic structural diagram of a scanning flow cytometric imaging analyzer in Example 2 of the present invention;

[0038] Figure 7 is a schematic structural diagram of a scanning galvanometer in Embodiment 3 of the present invention;

[0039] Figure 8 This is a schematic diagram of the structural principle of the liquid flow focusing module in Example 4 of the present invention;

[0040] Fig. 9 is a schematic diagram of a liquid flow focusing module in Example 5 of the present invention;

[0041] Fig.10 Schematic diagram of the focusing principle of the liquid flow focusing module in Example 5 of the present invention.

[0042] Description of reference numerals:

[0043] 1—Liquid flow focusing module; 2—Main optical path unit; 3—Laser emission optical path; 4—Optical signal detection optical path; 5—Scanning galvanometer; 6—Acquisition and control board; 7—Upper computer; 8—Velocity sensor; 9—Particles or cells; 10—Sample flow channel; 11—Sheath liquid flow channel; 12—First piezoelectric element; 13—Second piezoelectric element; 20—First cylindrical lens; 21—Second cylindrical lens; 22—Excitation pinhole; 23—First dichroic mirror; 24—Objective lens; 25—Detection pinhole; 2 6—collecting mirror; 30—laser; 31—first laser; 32—reflecting mirror; 33—second laser; 34—second dichroic mirror; 35—third laser; 36—third dichroic mirror; 40—interference filter; 41—lens; 42, 46, 47, 48—photomultiplier tube; 43—fourth dichroic mirror; 44—fifth dichroic mirror; 50—bracket; 51—elastic arm; 52—reflecting plate; 53—side arm; 54—piezoelectric plate; 55—mounting plate. DETAILED DESCRIPTION

[0044] The present invention is further described in detail below in conjunction with embodiments so that those skilled in the art can implement the invention with reference to the description.

[0045] It should be understood that the terms such as “having”, “including” and “comprising” used herein do not exclude the existence or addition of one or more other elements or combinations thereof.

[0046] Reference Figure 1 The present invention provides a scanning flow cytometer imaging analyzer, comprising:

[0047] A liquid flow focusing module 1, which provides a sample flow channel 10 and enables particles or cells 9 to flow in a single column in the sample flow channel 10 through focusing;

[0048] An optical signal excitation detection module, which is used to generate a laser spot and focus it on the detection area of ​​the sample flow channel 10 to excite the particles or cells 9 flowing through the detection area to generate fluorescence and scattered light, and to collect the fluorescence and scattered light signals generated by the particles or cells 9, and convert the optical signal into an electrical signal;

[0049] A scanning galvanometer 5, which is used to make the laser spot generated by the light signal exciting the detection module to perform reciprocating scanning along the orthogonal direction of the streamline of the particles or cells 9 in the detection area;

[0050] The acquisition and control board 6 receives the electrical signal transmitted by the optical signal exciting the detection module;

[0051] And the host computer 7 receives the signal transmitted by the acquisition and control board 6 and performs imaging.

[0052] In a preferred embodiment, the acquisition and control board 6 can also control the scanning galvanometer 5. The acquisition and control board 6 can be conventional, for example, NI PXI series board.

[0053] The laser spot generated by the optical signal excitation detection module is smaller than the size of the particle or cell 9 to be detected, so as to satisfy the point-by-point imaging of different regions of the particle or cell 9. When the cell flows through the detection area, the laser spot generates a scan perpendicular to the cell flow direction, such as Figure 2 As shown, it is a schematic diagram of the principle of laser spot scanning particles or cells 9. In a preferred embodiment, a velocity sensor 8 (conventional products can be selected) is also provided on the sample flow channel 10 to measure the velocity of particles or cells 9 in the sample flow channel 10. Since the particles or cells 9 flow in a straight line in a certain direction, coupled with the vertical scanning of the laser light spot, a plane scanning effect of point-by-point imaging can be formed. By measuring the cell flow velocity, combining the scanning frequency, and then superimposing and synthesizing the image data through the data acquisition and processing chip, a cell image is finally formed. Among them, it should be understood that the host computer 7 adopts a computer or a tablet computer and other equipment, which can perform imaging according to the signal transmitted by the acquisition and control board 6 through a conventional scheme. In the present invention, there is no specific limitation on the specific imaging scheme.

[0054] The scanning galvanometer 5 makes the laser spot scan back and forth perpendicular to the flow direction of the particles or cells 9, and the scanning period of the scanning galvanometer 5 is not greater than the ratio of the length of the spot on the particle / cell streamline to the flow velocity of the particles / cells.

[0055] The above is the overall concept of the present invention, and the present invention will be further described below in combination with more detailed embodiments based on the overall concept of the present invention.

[0056] Example 1

[0057] Reference Figure 1 and Figure 3 In this embodiment, the optical signal excitation detection module includes a main optical path unit 2, a group of laser emission optical paths 3, and a group of optical signal detection optical paths 4 corresponding to the laser emission optical paths 3;

[0058] The laser emission optical path 3 generates laser and transmits it to the main optical path unit 2. The main optical path unit 2 forms a laser spot and focuses on the particles or cells 9 in the detection area. The main optical path unit 2 also collects the fluorescence and scattered light generated by the particles or cells 9 in the detection area and transmits it to the optical signal detection optical path 4. The optical signal detection optical path 4 collects the fluorescence and scattered light signals and converts them into electrical signals and then transmits them to the acquisition and control board 6.

[0059] The main optical path unit 2 includes an excitation pinhole 22, a first dichroic mirror 23 and an objective lens 24 arranged in sequence along the incident direction of the laser, and a detection pinhole 25 arranged on the emission side of the fluorescence and scattered light of the objective lens 24. In a preferred embodiment, the light-incoming side of the excitation pinhole 22 is further provided with a first cylindrical lens 20 and a second cylindrical lens 21, and the emission side of the detection pinhole 25 also includes a collecting mirror 26;

[0060] The laser emission optical path 3 includes a laser 30;

[0061] The optical signal detection optical path 4 includes an interference filter 40, a lens 41 and a photomultiplier tube 42.

[0062] In this embodiment, the overall optical path is:

[0063] The laser emitted by the laser emission optical path 3 passes through the first cylindrical lens 20, the second cylindrical lens 21, and the excitation pinhole 22 in sequence to form a laser spot, which is then reflected by the first dichroic mirror 23 to the scanning galvanometer 5, and then focused by the objective lens 24 onto the particles or cells 9 in the detection area;

[0064] The fluorescence and scattered light generated by the particles or cells 9 in the detection area are collected by the objective lens 24, pass through the first dichroic mirror 23, and then pass through the detection pinhole 25 and the collection mirror 26 in sequence, and then enter the optical signal detection optical path 4;

[0065] The light entering the optical signal detection optical path 4 passes through the interference filter 40 and the lens 41 in sequence and then reaches the photomultiplier tube 42 . The photomultiplier tube 42 converts the optical signal into an electrical signal and transmits it to the acquisition and control board 6 .

[0066] In this embodiment, the scanning laser and the fluorescence collection share an objective lens 24. The focus of the objective lens 24 is the focal point of the scanning laser and is also the object point of instantaneous imaging. After the analyzer is focused, the scanning is limited to a plane of the sample. Only the light in the focal plane can pass through the detection pinhole 25. The light from the area outside the focal plane is defocused at the detection pinhole plane and cannot pass through the pinhole.

[0067] In this embodiment, a laser beam is used as a light source. The laser beam forms a laser spot after passing through the first cylindrical lens 20, the second cylindrical lens 21, and the excitation pinhole 22, and then is reflected to the scanning galvanometer 5 through the first dichroic mirror 23, and then is focused on the sample through the objective lens 24 to scan each point on the focal plane of the sample. If there is a fluorescent substance that can be excited in the sample, the fluorescence emitted after being excited directly passes through the first dichroic mirror 23 in the reverse direction through the original incident light path, and then passes through the detection pinhole 25 and the collection mirror 26 in turn, and is detected and collected by the photomultiplier tube 42 (PMT), and the signal is transmitted to the acquisition and control board 6, and then transmitted to the host computer 7, and imaged and displayed on the host computer 7.

[0068] Example 2

[0069] Reference Figure 4-6 As a further improvement on the basis of Example 1, in this embodiment, the laser emission optical path 3 and the optical signal detection optical path 4 can be expanded into multiple groups. When multiple lasers are used, multiple lasers can be used at the same point or multiple laser points can be separated by a certain spatial distance; at this time, the corresponding detection pinholes 25 also present a certain spatial arrangement, corresponding one to one with the multiple lasers; refer to Figure 4 , it is important to understand that Figure 4 From the perspective of the direction, the three laser beams overlap, but in fact they do not overlap. There is a certain distance between them, and this distance is reflected in another direction: the direction perpendicular to the paper surface.

[0070] In this embodiment, the number of laser emission optical paths 3 is N, and N is greater than 1. The laser emission optical path 3 further includes a reflector 32 or a dichroic mirror, so that the lasers emitted by the N lasers 30 are all introduced into the first cylindrical mirror 20 through the reflector 32 or the dichroic mirror; the number of optical signal detection optical paths 4 is M, and the optical signal detection optical path 4 further includes a dichroic mirror, so that the light of M different wavelength ranges emitted by the collecting mirror 26 is introduced into M different photomultiplier tubes 42 through the dichroic mirror. Generally speaking, one laser emission optical path 3 corresponds to multiple optical signal detection optical paths 4, such as N=1, M=3 or 4...; N=2, M=4 or 6..., etc.

[0071] Reference Figure 4 , in a more specific embodiment, N=3, i.e., including 3 laser emission light paths 3;

[0072] The first of the three laser emission optical paths 3 includes: a first laser 31 and a reflector 32, the second includes a second laser 33 and a second dichroic mirror 34, and the third includes a third laser 35 and a third dichroic mirror 36; the laser emitted by the first laser 31 is reflected by the reflector 32 and then sequentially transmits the second dichroic mirror 34 and the third dichroic mirror 36 to reach the first cylindrical mirror 20, the laser emitted by the second laser 33 is reflected by the second dichroic mirror 34 and then transmits the third dichroic mirror 36 to reach the first cylindrical mirror 20, and the laser emitted by the third laser 35 is reflected by the third dichroic mirror 36 to the first cylindrical mirror 20, so that all three lasers enter the main optical path.

[0073] Reference Figure 5 and Figure 6 In a more specific embodiment, M=3, that is, a three-way optical signal detection optical path 4, the three-way optical signal detection optical path 4 also includes two dichroic mirrors: a fourth dichroic mirror 43 and a fifth dichroic mirror 44. Among the light emitted by the collecting mirror 26, a part of the light is directly reflected by the fourth dichroic mirror 43 to a photomultiplier tube 46, a part of the light is transmitted through the fourth dichroic mirror 43 and then reflected by the fifth dichroic mirror 44 to another photomultiplier tube 47, and the remaining light is sequentially transmitted through the fourth dichroic mirror 43 and the fifth dichroic mirror 44 and then reflected to the remaining photomultiplier tube 48. Since there is an interference filter 40 in front of the photomultiplier tube 48, light of different wavelengths is collected, thereby realizing the collection and detection of three-way light.

[0074] Example 3

[0075] Reference Figure 7 As a further improvement on the basis of Example 1 or Example 2, in this embodiment, the scanning galvanometer 5 includes a bracket 50 with an opening in the middle, a reflector 52 suspended in the opening of the bracket 50 by two symmetrically distributed elastic arms 51, and a piezoelectric sheet 54 arranged on the side arm 53 of the bracket 50. The piezoelectric sheet 54 is used to generate an inverse piezoelectric effect under the action of an electric field, thereby driving the reflector 52 to twist and rotate. In this embodiment, a mounting sheet 55 is also provided on the bracket 50. The smaller the size of the scanning galvanometer 5, the higher the scanning frequency that can be achieved. For scanning above MHz, a micrometer-scale galvanometer can be manufactured using MEMS technology.

[0076] Example 4

[0077] Reference Figure 8 As a further improvement based on Example 1 or Example 2 or Example 3, in this embodiment, the liquid flow focusing module 1 includes a sample flow channel 10 and a sheath liquid flow channel 11 arranged outside the sample flow channel 10. The sheath liquid in the sheath liquid flow channel 11 produces an effect of wrapping the sample. Under the compression of the sheath liquid, the particles or cells 9 in the sample flow channel 10 are in a single-row flow state.

[0078] Example 5

[0079] Reference Fig. 9 and 10 As a further improvement based on Example 1 or Example 2 or Example 3, in this embodiment, the liquid flow focusing module 1 includes a sample flow channel 10, a first piezoelectric element 12 arranged on the sample flow channel 10 for achieving focusing of particles or cells 9 in the plane of the sample flow channel 10, and a second piezoelectric element 13 arranged on the sample flow channel 10 for achieving focusing of particles or cells 9 in the depth direction of the sample flow channel 10. In this embodiment, the piezoelectric element is a piezoelectric transducer.

[0080] Reference Fig.10 , is a schematic diagram of the focusing principle of the first piezoelectric element 12 and the second piezoelectric element 13. The wavelength of the ultrasonic standing wave generated by the first piezoelectric element 12 is about twice the width of the flow channel, and a standing wave node line is formed within the flow channel width range, and the node line position is close to the midline of the flow channel width. The particles or cells 9 move along the center line in the width direction under the continuous action of the sound wave, and the first piezoelectric element 12 achieves focusing in the XY plane; the wavelength of the ultrasonic standing wave generated by the second piezoelectric element 13 is about twice the depth of the flow channel, and a standing wave node line is formed within the flow channel depth range, and the node line position is close to the midline of the flow channel depth. The particles or cells 9 move along the center line in the depth direction under the continuous action of the sound wave, and the second piezoelectric element 13 achieves focusing in the YZ plane; under the action of the first and second piezoelectric elements 12 and 13, the particles or cells 9 move in a single row along the length direction of the flow channel at an equal speed, and there is no overtaking phenomenon, which provides favorable conditions for passing through the downstream laser detection area one by one, such as Figure 7 shown.

[0081] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to specific details.

Claims

1. A scanning flow cytometer, characterized in that: include: A liquid flow focusing module, which provides a sample flow channel and enables particles or cells to flow in a single file in the sample flow channel through focusing; An optical signal excitation detection module, which is used to generate a laser spot and focus it on the detection area of ​​the sample flow channel to excite the particles or cells flowing through the detection area to generate fluorescence and scattered light, and to collect the fluorescence and scattered light signals generated by the particles or cells, and convert the optical signal into an electrical signal; A scanning galvanometer, which is used to make the laser spot generated by the detection module excited by the optical signal to perform reciprocating scanning along the orthogonal direction of the particle or cell streamline in the detection area; An acquisition and control board, which receives the electrical signal transmitted by the optical signal excitation detection module; And a host computer, which receives the signal transmitted by the acquisition and control board and performs imaging.

2. The scanning flow cytometric imaging analyzer according to claim 1, characterized in that: The laser spot generated by the light signal excitation detection module is smaller than the size of the particles or cells to be detected, so as to satisfy the point-by-point imaging of different regions of the particles or cells.

3. The scanning flow cytometric imaging analyzer according to claim 2, characterized in that: The scanning galvanometer makes the laser spot scan back and forth perpendicular to the flow direction of the particles or cells, and the scanning period of the scanning galvanometer is not greater than the ratio of the length of the spot on the particle / cell streamline to the flow velocity of the particles / cells.

4. The scanning flow cytometric imaging analyzer according to claim 1, characterized in that: The optical signal excitation detection module includes a main optical path unit, at least one group of laser emission optical paths, and at least one group of optical signal detection optical paths corresponding to the laser emission optical paths; The laser emission optical path generates laser and transmits it to the main optical path unit. The main optical path unit forms a laser spot and focuses it on the particles or cells in the detection area. The main optical path unit also collects the fluorescence and scattered light generated by the particles or cells in the detection area and transmits it to the optical signal detection optical path. The optical signal detection optical path collects fluorescence and scattered light signals and converts them into electrical signals and then transmits them to the acquisition and control board.

5. The scanning flow cytometric imaging analyzer according to claim 4, characterized in that: The main optical path unit comprises a first cylindrical lens, a second cylindrical lens, an excitation pinhole, a first dichroic mirror and an objective lens which are sequentially arranged along the incident direction of the laser, and a detection pinhole and a collecting mirror which are sequentially arranged along the optical path direction of the fluorescence and scattered light emitted by the objective lens; The laser emission optical path includes a laser; The optical signal detection optical path includes an interference filter, a lens and a photomultiplier tube. The light entering the optical signal detection optical path passes through the interference filter and the lens in sequence and then reaches the photomultiplier tube. The photomultiplier tube converts the optical signal into an electrical signal and transmits it to the acquisition and control board.

6. The scanning flow cytometric imaging analyzer according to claim 5, characterized in that: The laser emitted by the laser emission optical path passes through the first cylindrical lens, the second cylindrical lens, and the excitation pinhole in sequence to form a laser spot, which is then reflected by the first dichroic mirror to the scanning galvanometer mirror and then focused by the objective lens onto the particles or cells in the detection area; The fluorescence and scattered light generated by the particles or cells in the detection area are collected by the objective lens, pass through the first dichroic mirror, and then pass through the detection pinhole and the collecting mirror in sequence, and then enter the optical signal detection optical path.

7. The scanning flow cytometric imaging analyzer according to claim 6, characterized in that: The number of the laser emission optical paths is N, and N is greater than 1. The laser emission optical paths further include a reflector or a dichroic mirror, so that the lasers emitted by the N lasers are all guided into the first cylindrical mirror through the reflector or the dichroic mirror; The number of the optical signal detection optical paths is M, and the optical signal detection optical paths further include a dichroic mirror, so as to guide the light of M different wavelength ranges emitted by the collecting mirror into M different photomultiplier tubes through the dichroic mirror.

8. The scanning flow cytometric imaging analyzer according to any one of claims 1 to 7, characterized in that: The scanning galvanometer includes a bracket with an opening in the middle, a reflector suspended in the opening of the bracket by two symmetrically distributed elastic arms, and a piezoelectric sheet arranged on the side arm of the bracket. The piezoelectric sheet is used to generate an inverse piezoelectric effect under the action of an electric field, thereby driving the reflector to twist.

9. The scanning flow cytometric imaging analyzer according to any one of claims 1 to 7, characterized in that: The liquid flow focusing module includes a sample flow channel and a sheath liquid flow channel arranged at the periphery of the sample flow channel. The sheath liquid in the sheath liquid flow channel causes the particles or cells in the sample flow channel to present a single-row flow state.

10. The scanning flow cytometric imaging analyzer according to any one of claims 1 to 7, characterized in that: The liquid flow focusing module includes a sample flow channel, a first piezoelectric element arranged on the sample flow channel for focusing particles or cells within the plane of the sample flow channel, and a second piezoelectric element arranged on the sample flow channel for focusing particles or cells in the depth direction of the sample flow channel.

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