An intelligent microthrombus detection and analysis system and analysis instrument

Abstract

The present invention belongs to the field of medical detection technology, and particularly relates to an intelligent microthrombus detection and analysis system and an analysis instrument, including a multimodal imaging optical path, a microfluidic device and a data processing module. The imaging optical path is composed of a continuous laser light source, a first beam splitter, a first reflector, a second reflector, a third reflector, a fourth reflector, a first acousto-optic modulator, a second acousto-optic modulator, a first beam combining mirror, a first convex lens, a second convex lens, a first objective lens, a microfluidic chip, a second objective lens, a third convex lens, a first photodetector, an arbitrary wave generator, a data acquisition card, a fourth convex lens, a fifth convex lens, a fifth reflector, a sixth reflector, a second beam combining mirror, a dichroic mirror, a sixth convex lens, and a second photodetector; the microfluidic device is composed of a microfluidic chip and an injection pump; and the data processing module is composed of a computer integrated with a data acquisition card and a data processing program. Compared with the existing technology, the present invention is based on acousto-optic modulation and frequency division multiplexing technology, which can obtain multi-dimensional information such as the number, concentration, morphology, density, composition, biochemical activity, etc. of microthrombi, and realize high-speed, high-throughput, and high-resolution microthrombus detection and analysis.

Description

Technical Field

[0001] The present invention belongs to the field of medical detection technology, and in particular relates to an intelligent microthrombus detection and analysis system and an analysis instrument. Background Art

[0002] Microthrombi are precursors of thrombi, with a size of less than 100 microns, and often appear in small veins, capillaries and microcirculation. The appearance of microthrombi is considered to be an important manifestation of early cardiovascular and cerebrovascular diseases. The detection and analysis of microthrombi is the key to early diagnosis, pathological understanding, and early precision treatment of cardiovascular and cerebrovascular diseases, which helps to improve the cure rate of the disease and reduce mortality, disability and treatment costs. Clinically, medical imaging technologies (such as tomography, nuclear magnetic resonance, ultrasound, etc.) can directly observe thrombi or vascular stenosis caused by thrombi, but due to the limited spatial resolution of these medical imaging technologies, it is impossible to detect and analyze microthrombi in the blood circulation of early patients before macroscopic large-scale thrombi are formed, and the diagnosis timeliness is poor.

[0003] Traditional optical microscopes and flow cytometers are the most important and common tools for detecting and analyzing microthrombi and platelets (the most important blood cells involved in coagulation and thrombosis). Optical microscopes have extremely high spatial resolution and can directly observe the morphology of microthrombi, and are directly used to determine the size and composition of microthrombi and the degree of platelet activation. However, the detection speed of traditional optical microscopes is too slow, and it requires a lot of manpower to observe and rely on empirical analysis. It is impossible to perform large-scale, quantitative, and multidimensional detection and analysis of microthrombi and platelets in a statistically significant way, and can only provide qualitative results. Flow cytometry measures the physicochemical properties of microthrombi and platelets (such as impedance, light scattering, fluorescence, etc.) and performs large-scale detection and analysis in a statistically significant way. However, due to its lack of spatial resolution, it is impossible to distinguish between microthrombi and single platelets, nor can it determine the degree of platelet activation. The optical turbidimetry of the coagulometer is a method widely used in clinical testing. It detects the average in vitro aggregation characteristics of a large number of platelets by measuring optical transmittance. However, it cannot give the quantitative distribution of the size of microthrombi formed during the aggregation process and the proportion of microthrombi formation, making it difficult to perform statistical analysis.

[0004] In recent years, optofluidic imaging technology combines the high-resolution advantages of traditional optical microscopes with the high-throughput detection advantages of flow cytometers, making it possible to achieve large-scale, quantitative microthrombi and platelet detection and analysis. In 2011, Professor Jean Feuillard and others from the University of Limoges in France first used optofluidic imaging technology to study microthrombi and platelets. They used optofluidic fluorescence imaging technology based on traditional high-speed charge-coupled device (CCD) cameras to observe the effect of platelet attachment to the surface of leukocytes on fluorescent labeling, and quantitatively gave the changes in the number of microthrombi formed by platelet and leukocyte aggregation under different experimental conditions. In 2015, Professor Matthew Linden and others from the University of Western Australia also used optofluidic fluorescence imaging technology based on CCD cameras to observe microthrombi formed by platelet and monocyte aggregation, demonstrating the accuracy advantage of imaging-based flow cytometers over traditional non-imaging flow cytometers in counting. In 2017, Professor Matthew Linden and others used optofluidic fluorescence imaging technology based on CCD cameras to further observe microthrombi formed by the aggregation of platelets and various white blood cells. In the same year, Professor Keisuke Goda and others from the University of Tokyo in Japan used ultrafast optofluidic bright-field imaging technology based on time-domain stretching to greatly improve the detection throughput, and used machine learning to identify image information to achieve successful classification of single platelets, white blood cells and microthrombi. In 2020, Professor Lei Cheng and others from Wuhan University also used ultrafast optofluidic bright-field imaging technology based on time-domain stretching to achieve classification and identification of microthrombi under the action of different agonists. In 2022, Professor Paul Harrison and others from the University of Birmingham in the UK also used optofluidic fluorescence imaging technology based on CCD cameras to achieve identification and analysis of proplatelet and barbell platelet microthrombi. In 2023, Denese Marks and others from the University of Sydney in Australia used optofluidic multicolor fluorescence imaging technology based on CCD cameras to achieve identification and classification of platelet subpopulations and microthrombi.

[0005] However, the current optofluidic imaging technology used to analyze microthrombi and platelets is still mostly based on high-speed CCD cameras. Due to the imaging speed of high-speed CCD cameras, the detection throughput is limited to 10 per second. 3 In addition, since CCD cameras have no imaging selectivity, it is impossible to image only microthrombi and platelets for whole blood samples. The effective detection flux is usually less than 10 cell objects per second, which is difficult to use for large-scale actual clinical sample detection. Although ultrafast laser optofluidic imaging technology based on time domain stretching can increase the effective detection flux to 10 per second, it is still difficult to use ultrafast laser optofluidic imaging technology based on time domain stretching. 4However, due to the limitation of previous imaging principles, only single image information can be obtained. Therefore, it is urgent to develop a microthrombus detection and analysis technology and instrument to analyze the microthrombi and platelets in the blood circulation of patients with early cardiovascular and cerebrovascular diseases in a large-scale, quantitative, and multi-dimensional manner (including quantity, concentration, morphology, density, composition, biochemical activity, etc.). Summary of the invention

[0006] The purpose of the present invention is to provide an intelligent microthrombus detection and analysis system and analysis instrument to address the technical bottleneck of existing optofluidic imaging technology in large-scale microthrombus detection. The present invention is based on acousto-optic modulation and frequency division multiplexing technology, and can achieve high-speed, high-resolution optofluidic imaging including simultaneous imaging of four modes including bright field, quantitative phase, fluorescence, and fluorescence lifetime. Compared with traditional optofluidic imaging technology, the present invention can obtain bright field images, quantitative phase images, fluorescence images, and fluorescence lifetime images of microthrombi respectively. These four images contain information on different dimensions of microthrombi. Bright field images can provide information on the number, concentration, morphology, and other aspects of microthrombi in blood samples. Quantitative phase images can provide information on the density and composition of microthrombi. Fluorescence and fluorescence lifetime images can provide information on the composition and biochemical activity of microthrombi.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is:

[0008] An intelligent microthrombus detection and analysis system comprises a multimodal imaging optical path, a microfluidic device and a data processing module; the multimodal imaging optical path comprises a bright field imaging optical path, a quantitative phase imaging optical path, a fluorescence imaging optical path and a fluorescence lifetime imaging optical path; the microfluidic device is arranged in the multimodal imaging optical path for collecting microthrombus information, and the data processing module is used for analyzing and processing the optical signal data in the multimodal imaging optical path; the microfluidic device is composed of a microfluidic chip, an injection tube, a syringe and an injection pump connected together; the data processing module comprises a photodetector, a data acquisition card and a data processing program.

[0009] Furthermore, the bright field imaging optical path includes a continuous laser light source, the continuous laser emitted by the continuous laser light source is divided into a first laser beam and a second laser beam by a first beam splitter, the first laser beam passes through a first reflector (on the premise that the optical path of the first laser beam and the second laser beam from after the first beam splitter to before the first beam combining mirror is the same, and the number of reflectors is adjusted according to the laboratory conditions) and enters the first acousto-optic modulator to be modulated and diffracted, the second laser beam passes through a fourth reflector and enters the second acousto-optic modulator to be modulated and diffracted, and also includes an arbitrary wave generator, the waves output by the arbitrary wave generator enter the first acousto-optic modulator and the second acousto-optic modulator respectively and modulate the first laser beam and the second laser beam respectively, the modulated first laser beam and the second laser beam interfere in the first beam combining mirror and are again divided into two beams, one of which passes through the first convex lens and the second convex lens for collimation and beam expansion, is focused by the first objective lens into a spot array, irradiates the microfluidic chip as signal light, is collected by the second objective lens, and is then focused by the third convex lens to the first photodetector and transmitted to the data acquisition card to collect electrical signals and form an image, and the other beam is used as a reference optical path.

[0010] Furthermore, the quantitative phase imaging optical path includes a bright field imaging optical path and a reference optical path, wherein the reference optical path includes a fourth convex lens, a fifth convex lens, a fifth reflector, a sixth reflector and a second beam combining mirror connected in sequence to the first beam combining mirror optical path, and the second beam combining mirror is arranged on the optical path between the second objective lens and the third convex lens.

[0011] Furthermore, the fluorescence imaging optical path and the fluorescence lifetime imaging optical path include a bright field imaging optical path, and also include a dichroic mirror arranged between the second convex lens of the bright field imaging optical path and the second objective lens optical path, and the dichroic mirror feeds back the light signal to the second photodetector through the sixth convex lens.

[0012] Furthermore, the wavelength of the continuous laser light source is 193 nm to 20 μm, preferably 491 nm; the splitting ratio of the first beam splitter is in the range of 1:9 to 9:1, preferably 5:5.

[0013] Furthermore, the dichroic mirror has a transmittance of >99% for the wavelength of the continuous laser light source and a reflectance of >99% for the wavelength of the fluorescence.

[0014] Furthermore, the first photodetector and the second photodetector are avalanche photodetectors.

[0015] Furthermore, the microfluidic chip is placed on the image-side focal plane of the first objective lens and on the object-side focal plane of the second objective lens. The magnifications of the first objective lens and the second objective lens range from 5 to 100 times, and the numerical apertures range from 0.1 to 1.48. Preferably, the magnification of the first objective lens is 20 times, and the numerical aperture is 0.80, and the magnification of the second objective lens is 20 times, and the numerical aperture is 0.40.

[0016] An intelligent microthrombus detection and analysis instrument uses the above-mentioned intelligent microthrombus detection and analysis system to realize its analysis function.

[0017] Working principle: The specific working mode of the multimodal imaging optical path is as follows: the continuous laser emitted by the continuous laser light source is divided into two laser beams (hereinafter referred to as the first laser beam and the second laser beam) through the first beam splitter. The first laser beam passes through the first reflector, the second reflector and the third reflector to enter the first acousto-optic modulator to be modulated and diffracted, and the second laser beam passes through the fourth reflector to enter the second acousto-optic modulator to be modulated and diffracted. The arbitrary wave generator emits a wave of a specific frequency and inputs it into the first acousto-optic modulator and the second acousto-optic modulator. The two modulated and diffracted light beams generate two sets of spatially dispersed optical frequency combs with intensity modulation through the beat frequency. The two sets of optical frequency combs have the same diffraction order and the same frequency. The light beams interfere in the first beam combiner and are divided into two beams again. One of the beams is collimated and expanded by the first convex lens and the second convex lens, and then focused by the first objective lens into a spot array to irradiate the microfluidic channel in the microfluidic chip as the signal light. When the microthrombus flows through the focused spot array in the microfluidic channel, the information of each point of the microthrombus is detected by the focused spot array. The signal light carrying the microthrombus information is collected by the second objective lens and focused by the third convex lens into the first photodetector, converting the optical signal into an electrical signal, and finally the data acquisition card collects the electrical signal for bright field imaging; another beam of reference light is collimated and expanded by the fourth and fifth convex lenses, and then enters the second beam combiner through the fifth and sixth reflectors to interfere with the signal light, and then is focused by the third convex lens into the first photodetector to convert the optical signal into an electrical signal, and finally the data acquisition card collects the electrical signal for quantitative phase imaging. At the same time, the backscattered fluorescence of each point of the excited microthrombus is collected by the first objective lens, passes through the dichroic mirror, and is focused by the sixth convex lens into the second photodetector for fluorescence and fluorescence lifetime imaging. The specific working method of the microfluidic device is as follows: the microfluidic chip is placed on the image focal plane of the first objective lens and the object focal plane of the second objective lens in the multimodal imaging optical path. The microfluidic chip leads to two injection tubes, one of which is connected to a syringe containing microthrombi, which is installed in a syringe pump. The syringe pump pushes the syringe at a specific speed to inject the microthrombi into the microfluidic channel of the microfluidic chip, and the other injection tube is connected to a waste liquid bucket to recover the microthrombus samples. The specific working mode of the data processing module is as follows: the data acquisition card transmits the electrical signal data collected by the photodetector in the multimodal imaging optical path to the data processing program, and the data processing program uses Fourier transform to convert the time domain signal measured by the photodetector into a frequency domain signal, so that the transmission intensity, interference intensity, fluorescence intensity and phase delay of each point of the microthrombus can be obtained, thereby reconstructing the bright field image, quantitative phase image, fluorescence image and fluorescence lifetime image of the microthrombus respectively.

[0018] The advantages of the present invention are: based on acousto-optic modulation and frequency division multiplexing (specifically, multiple frequency signals use the same channel. Each light comb after beam combining has a different frequency signal. After irradiating the sample in the microfluidic chip, the intensity of each comb changes, that is, the intensity of each frequency is different. After focusing, the light comb enters the same channel, that is, the first photodetector, which can greatly speed up the imaging speed) technology, the present invention can realize high-speed, high-resolution optofluidic imaging including simultaneous imaging of four modes including bright field, quantitative phase, fluorescence, and fluorescence lifetime. Compared with traditional optofluidic imaging technology, the present invention can obtain bright field images, quantitative phase images, fluorescence images, and fluorescence lifetime images of microthrombi respectively. These four images contain information of different dimensions of microthrombi. Bright field images can provide information such as the number, concentration, and morphology of microthrombi in blood samples, quantitative phase images can provide information such as the density and components of microthrombi, and fluorescence and fluorescence lifetime images can provide information such as the components and biochemical activity of microthrombi. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a schematic diagram of the bright field imaging structure of the intelligent microthrombus detection and analysis system provided in Example 1 of the present invention.

[0020] Figure 2 It is a schematic diagram of the quantitative phase imaging structure of the intelligent microthrombus detection and analysis system provided in Example 2 of the present invention.

[0021] Figure 3 It is a schematic diagram of the fluorescence and fluorescence lifetime imaging structure of the intelligent microthrombus detection and analysis system provided in Example 3 of the present invention.

[0022] Figure 4 It is a schematic diagram of the multimodal imaging structure of the intelligent microthrombus detection and analysis system of Example 4 of the present invention.

[0023] Figure 5 This is a diagram of the detection results of platelets using the intelligent microthrombus detection and analysis system of the present invention, in which a is a bright field image of a single platelet, b is a bright field image of a large number of platelet aggregations, and c is a bright field image of a large number of platelets and white blood cells aggregations.

[0024] Among them, 1-continuous laser light source, 2-first beam splitter, 3-first reflector, 4-second reflector, 5-third reflector, 6-fourth reflector, 7-first acousto-optic modulator, 8-second acousto-optic modulator, 9-first beam combining mirror, 10-first convex lens, 11-second convex lens, 12-first objective lens, 13-microfluidic chip, 14-second objective lens, 15-third convex lens, 16-first photodetector, 17-arbitrary wave generator, 18-data acquisition card, 19-fourth convex lens, 20-fifth convex lens, 21-fifth reflector, 22-sixth reflector, 23-second beam combining mirror, 24-dichroic mirror, 25-sixth convex lens, 26-second photodetector. DETAILED DESCRIPTION

[0025] Example 1

[0026] like Figure 1 As shown, an intelligent microthrombus detection and analysis system includes a bright field imaging optical path, a microfluidic device and a data processing module; the microfluidic device is arranged in the bright field imaging optical path for collecting microthrombus information, and the data processing module is used to analyze and process the optical signal data in the multimodal imaging optical path; the microfluidic device includes a microfluidic chip and a syringe pump, and the microfluidic chip is placed on the image focal plane of the first objective lens and on the object focal plane of the second objective lens; the data processing module includes a photodetector, a data acquisition card and a data processing program. The bright field imaging optical path includes a continuous laser light source 1. The continuous laser emitted by the continuous laser light source is divided into a first laser beam and a second laser beam by a first beam splitter 2. The first laser beam passes through a first reflector 3, a second reflector 4, and a third reflector 5 in sequence (presumably, the optical paths of the first laser beam and the second laser beam from after the first beam splitter to before the first beam combiner are the same. Three reflectors are selected according to the laboratory conditions) and enters a first acousto-optic modulator 7 to be modulated and diffracted. The second laser beam passes through a fourth reflector 6 and enters a second acousto-optic modulator 8 to be modulated and diffracted. The bright field imaging optical path also includes an arbitrary wave generator 17. The arbitrary wave generator 17 outputs The outgoing waves enter the first acousto-optic modulator 7 and the second acousto-optic modulator 8 respectively to modulate the first laser beam and the second laser beam respectively. The modulated first laser beam and the second laser beam interfere in the first beam combiner 9 and are divided into two beams again, one of which is collimated and expanded by the first convex lens 10 and the second convex lens 11, and is focused by the first objective lens into a spot array to illuminate the microfluidic chip 13 as a signal light and collected by the second objective lens 14, and then focused by the third convex lens 15 to the first photodetector (avalanche photodetector) 16 and transmitted to the data acquisition card 18 to collect electrical signals and form an image, and the other beam is used as a reference light path.

[0027] Specifically, the wavelength of the continuous laser light source is 491 nm, which is used to emit continuous laser light. The splitting ratio of the first beam splitter is 50:50, which is used to split the continuous laser light into two beams of laser light with exactly the same power (hereinafter referred to as the first laser beam and the second laser beam). The first laser beam passes through the first reflector, the second reflector and the third reflector and enters the first acousto-optic modulator to be modulated and diffracted to generate an optical frequency comb with a frequency of 100 MHz to 200 MHz. The second laser beam passes through the fourth reflector and enters the second acousto-optic modulator to be modulated and diffracted to generate an optical frequency comb with a frequency of 200 MHz to 400 MHz. The arbitrary wave generator emits a wave of a specific frequency and enters the first acousto-optic modulator and the second acousto-optic modulator. The two modulated and diffracted light beams generate two sets of spatially dispersed optical frequency combs with a frequency of 200 MHz to 400 MHz with intensity modulation through the beat frequency. The two sets of optical frequency combs have the same diffraction order and the same frequency beams, which interfere in the first beam combiner and then merge into one optical frequency comb. The focal length of the first convex lens is 50 mm, and the focal length of the second convex lens is 60 mm. These two lenses collimate and expand the optical frequency comb. The magnification of the first objective lens is 20 times and the numerical aperture is 0.80, and the magnification of the second objective lens is 20 times and the numerical aperture is 0.40. The microfluidic chip is placed in the image focal plane of the first objective lens and the object focal plane of the second objective lens. After the optical frequency comb is focused on the microthrombus sample in the microfluidic channel of the microfluidic chip by the first objective lens, the information carrying each point of the microthrombus is collected by the second objective lens. The focal length of the third convex lens is 50 mm, which is used to focus the light beam carrying the information of each point of the microthrombus to the first photodetector. The bandwidth of the first photodetector is 200 MHz-400 MHz, which is used to convert the optical signal into an electrical signal. The acquisition speed of the data acquisition card is 1.2 GHz, which is used to collect electrical signals. The collected electrical signal contains the transmission intensity of each point of the microthrombus, so the microthrombus can be imaged in the bright field.

[0028] Example 2

[0029] The difference between Example 2 and Example 1 is that: on the basis of Example 1, an additional reference optical path is added, that is, the quantitative phase imaging optical path, on the basis of the bright field imaging optical path, also includes an optical path, namely, the fourth convex lens 19, the fifth convex lens 20, the fifth reflector 21, the sixth reflector 22 and the second beam combining mirror 23 which are connected in sequence to the optical path of the first beam combining mirror 9, and the second beam combining mirror is arranged on the optical path between the second objective lens and the third convex lens.

[0030] Specifically, a fourth convex lens, a fifth convex lens, a fifth reflector, a sixth reflector, and a second beam combiner are placed on the other side of the first beam combiner. The focal length of the fourth convex lens is 50 mm, and the focal length of the fifth convex lens is 60 mm. One of the two optical frequency combs passing through the first beam combiner is exactly the same as the optical path of Example 1, and the other enters the fourth convex lens and the fifth convex lens for collimation and beam expansion, and then passes through the fifth reflector and the sixth reflector to enter the second beam combiner to combine with the optical frequency comb carrying the information of each point of the microthrombus, and finally focuses through the third convex lens to enter the first photodetector, and the first photodetector converts the optical signal into an electrical signal. The data acquisition card has an acquisition speed of 1.2 GHz and is used to collect electrical signals. The electrical signal contains the interference intensity and phase information of each point of the microthrombus, so quantitative phase imaging of the microthrombus can be performed.

[0031] Example 3

[0032] The difference between Example 3 and the example is that: on the basis of Example 1, an additional fluorescence detection optical path is added, and the fluorescence imaging optical path and the fluorescence lifetime imaging optical path are based on the bright field imaging optical path, and also include a dichroic mirror 24 arranged between the second convex lens of the bright field imaging optical path and the second objective lens optical path, and the dichroic mirror feeds back the light signal to the second photodetector 26 through the sixth convex lens.

[0033] Specifically, a dichroic mirror, a sixth convex lens, and a second photodetector are placed behind the second convex lens. The backscattered fluorescence of each point of the excited microthrombus is collected by the first objective lens, and after passing through the dichroic mirror and being focused by the sixth convex lens, it enters the second photodetector to convert the optical signal into an electrical signal. The data acquisition card has an acquisition speed of 1.2 GHz and is used to collect electrical signals. The electrical signal contains the fluorescence intensity and phase delay of each point of the microthrombus, so fluorescence and fluorescence lifetime imaging of the microthrombus can be performed.

[0034] Example 4

[0035] Embodiment 4 is a multimodal imaging optical path system that integrates the bright field imaging optical path of embodiment 1, the quantitative phase imaging optical path of embodiment 2, the fluorescence imaging optical path and the fluorescence lifetime imaging optical path of embodiment 3. The bright field imaging optical path, the quantitative phase imaging optical path, the fluorescence imaging optical path and the fluorescence lifetime imaging optical path are integrated into a comprehensive multimodal intelligent microthrombus detection and analysis system, which can respectively obtain bright field images, quantitative phase images, fluorescence images and fluorescence lifetime images of microthrombi. These four images contain information of different dimensions of microthrombi. The bright field image can provide information such as the number, concentration, and morphology of microthrombi in blood samples. The quantitative phase image can provide information such as the density and components of microthrombi. The fluorescence and fluorescence lifetime images can provide information such as the components and biochemical activity of microthrombi. Based on acousto-optic modulation and frequency division multiplexing technology, multi-dimensional information such as the number, concentration, morphology, density, components, and biochemical activity of microthrombi can be obtained, thereby realizing high-speed, high-throughput, and high-resolution microthrombus detection and analysis.

Claims

1. An intelligent microthrombus detection and analysis system, characterized in that: It comprises a multimodal imaging optical path, a microfluidic device and a data processing module; the multimodal imaging optical path comprises a quantitative phase imaging optical path, a fluorescence imaging optical path and a fluorescence lifetime imaging optical path; the quantitative phase imaging optical path, the fluorescence imaging optical path and the fluorescence lifetime imaging optical path all comprise a bright field imaging optical path, the microfluidic device is arranged in the multimodal imaging optical path for collecting microthrombus information, and the data processing module is used for analyzing and processing the optical signal data in the multimodal imaging optical path; the microfluidic device comprises a microfluidic chip and an injection pump; the data processing module comprises a photodetector, a data acquisition card and a data processing program; the bright field imaging optical path comprises a continuous laser light source, and the continuous laser emitted by the continuous laser light source is divided into a first laser beam and a second laser beam by a first beam splitter, The first laser beam passes through the first reflector and enters the first acousto-optic modulator to be modulated and diffracted, and the second laser beam passes through the fourth reflector and enters the second acousto-optic modulator to be modulated and diffracted. An arbitrary wave generator is also included. The waves output by the arbitrary wave generator enter the first acousto-optic modulator and the second acousto-optic modulator respectively to modulate the first laser beam and the second laser beam respectively. The modulated first laser beam and the second laser beam interfere in the first beam combining mirror and are divided into two beams again, one of which passes through the first convex lens and the second convex lens for collimation and beam expansion, is focused by the first objective lens into a spot array, and is irradiated into the microfluidic chip as signal light and collected by the second objective lens, and then is focused by the third convex lens to the first photodetector and transmitted to the data acquisition card to collect electrical signals and form an image, and the other beam is used as a reference optical path.

2. The intelligent microthrombus detection and analysis system according to claim 1, characterized in that: The quantitative phase imaging optical path also includes a reference optical path, which includes a fourth convex lens, a fifth convex lens, a fifth reflector, a sixth reflector and a second beam combiner connected in sequence to the first beam combiner optical path, and the second beam combiner is arranged on the optical path between the second objective lens and the third convex lens.

3. The intelligent microthrombus detection and analysis system according to claim 1, characterized in that: The fluorescence imaging optical path and the fluorescence lifetime imaging optical path also include a dichroic mirror arranged between the second convex lens of the bright field imaging optical path and the second objective lens optical path, and the dichroic mirror feeds back the optical signal to the second photodetector through the sixth convex lens.

4. The intelligent microthrombus detection and analysis system according to any one of claims 1 to 3, characterized in that: The wavelength of the continuous laser light source is 193nm-20μm; the splitting ratio range of the first beam splitter is 1:9-9:

1.

5. The intelligent microthrombus detection and analysis system according to claim 4, characterized in that: The first laser beam and the second laser beam have the same optical path from after the first beam splitter to before the first beam combiner.

6. The intelligent microthrombus detection and analysis system according to claim 3, characterized in that: The dichroic mirror has a wavelength transmittance of >99% for the continuous laser light source and a wavelength reflectivity of >99% for the fluorescence.

7. The intelligent microthrombus detection and analysis system according to claim 6, characterized in that: The first photodetector and the second photodetector are avalanche photodetectors.

8. The intelligent microthrombus detection and analysis system according to claim 7, characterized in that: The microfluidic chip is placed on the image focal plane of the first objective lens and the object focal plane of the second objective lens. The magnification of the first objective lens and the second objective lens ranges from 5 to 100 times, and the numerical aperture ranges from 0.1 to 1.

48.

9. An intelligent microthrombus detection and analysis instrument, characterized in that: The instrument is implemented using the intelligent microthrombus detection and analysis system described in any one of claims 1-8.

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