Portable full-automatic fluorescence spectrum sperm activity detection method and system

Abstract

The application provides a portable full-automatic fluorescence spectrum sperm activity detection method and system, and belongs to the technical field of sperm activity detection. The method comprises the following steps: placing a semen sample to be detected into a detection kit to which a sperm activity fluorescence biological dyeing agent is added; the sperm activity fluorescence biological dyeing agent comprises a CM-Dil biological derivative activation dyeing solution; fluorescence spectrum information of the semen sample to be detected is acquired through a far-field optical fluorescence microscope and a microscopic optical information acquisition assembly; the fluorescence spectrum information is subjected to data analysis through a spectrum analysis system, so as to obtain survival rate information, development state information and activity classification information of sperm in the semen sample. The application realizes that a person to be detected can complete sperm activity detection by himself without going to a hospital; and the detection result contains the survival rate information, the development state information and the activity classification information of sperm in the semen sample, so that whether the person to be detected has the ability to reproduce can be accurately determined.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of sperm activity detection, and particularly relates to a portable full-automatic fluorescence spectrum sperm activity detection method and system. BACKGROUND

[0002] The sperm activity and morphology are key parts in human semen examination, and are very important for eugenics and scientific preparation for pregnancy. The existing sperm detection needs to be completed in a hospital. After an experimenter roughly judges the semen properties, whether there is liquefaction, whether it is viscous or dilute, and whether there are white blood cells and red blood cells, a computer is used for auxiliary analysis to obtain the sperm density, the number of sperm per milliliter, and the sperm activity of the semen sample. The sperm activity is classified into fast forward, slow forward, and immobile sperm, and the sperm deformity rate. However, the sperm activity detection in the hospital needs four steps of preparation before examination, sperm collection, semen analysis, and reading of the analysis report, and it takes at least one week to complete one detection. Moreover, the detection instrument and the experience of the doctor are required.

[0003] In the prior art, there is a method for detecting sperm activity by using a sperm motility density test paper. However, the sperm motility density test paper can only detect the number of sperm, and about 10% of the to-be-detected persons are determined as negative because the number of sperm does not reach the critical value of the sperm motility density test paper, but they still have certain fertility. Therefore, the sperm motility density test paper cannot be used as a method for determining whether the to-be-detected person has fertility.

[0004] Therefore, there is an urgent need for a convenient and efficient sperm activity detection method and system. SUMMARY

[0005] The application provides a portable full-automatic fluorescence spectrum sperm activity detection method, system, electronic equipment, and storage medium to overcome at least one technical problem in the prior art.

[0006] To achieve the above-mentioned purpose, the application provides a portable full-automatic fluorescence spectrum sperm activity detection method, which comprises the following steps: placing a to-be-detected semen sample into a detection kit added with a sperm activity fluorescence biological dyeing agent; the sperm activity fluorescence biological dyeing agent comprises a CM-Dil biological derivative activation dyeing solution.

[0007] Fluorescence spectrum information of the to-be-detected semen sample is acquired by a far-field optical fluorescence microscope and a microscopic optical information acquisition assembly.

[0008] Fluorescence spectral information is analyzed using a spectral analysis system to obtain information on sperm survival rate, developmental status, and motility grading in semen samples.

[0009] Furthermore, a preferred method is that the method for obtaining the CM-Dil bio-derived activated staining solution includes:

[0010] Add 10 mg of CM-Dil cell staining agent to 10 ml of HBSS buffer and stir to dissolve;

[0011] Then add 250 μL of EDC at a concentration of 2 mg / mL and 60 μL of NHS at a concentration of 2 mg / mL, and activate at 37°C for 30 min;

[0012] After the activated solution was reacted in a shaker at 37°C for 6 hours, the reaction product was purified to obtain the CM-Dil bio-derived activated staining solution.

[0013] Further, a preferred method is that the HBSS buffer comprises: 0.1-0.5 mol / L sodium chloride, 0.2-2 mmol / L ethylenediaminetetraacetic acid, 0.01-0.05 mol / L sodium dodecyl sarcosinate, 0.05-0.2 mol / L citric acid, and 0.1-0.5 mol / L disodium hydrogen phosphate.

[0014] Furthermore, a preferred method involves analyzing fluorescence spectral information using a spectral analysis system to obtain sperm viability, developmental status, and motility grading information from semen samples. This includes methods such as:

[0015] The fluorescence spectral information was preprocessed using a pre-defined sperm motility detection model to obtain denoised fluorescence spectral information;

[0016] The denoised fluorescence spectrum information was analyzed by a spectrometer to obtain information on sperm survival rate, developmental status, and motility grading of the semen sample.

[0017] Furthermore, a preferred method is that the spectrometer separates the electromagnetic radiation from the radiation source into the desired wavelength region using a dispersive element, and measures the intensity at the selected wavelength; the wavelength region includes: the wavelength region of blue light is 435-450 nm; the wavelength region of green light is 495-580 nm; and the wavelength region of red light is 645-760 nm.

[0018] Furthermore, a preferred method is that when the sperm activity fluorescent biological staining agent also includes acridine orange dye, in addition to obtaining information on the survival rate, developmental status, and motility grading of the semen sample, it can also obtain information on the DNA fragmentation rate of the sperm.

[0019] To address the aforementioned issues, this invention also provides a portable, fully automated fluorescence spectroscopy sperm motility detection system, comprising: a detection unit for placing a semen sample to be tested into a detection kit containing a sperm motility fluorescent biological staining agent; the sperm motility fluorescent biological staining agent includes CM-Dil biological derivative activation staining solution;

[0020] The spectral information acquisition unit is used to acquire the fluorescence spectral information of the semen sample to be tested through a far-field optical fluorescence microscope and a microscopic optical information acquisition component;

[0021] The data analysis unit is used to analyze fluorescence spectral information through a spectral analysis system to obtain information on sperm survival rate, developmental status, and motility grading of semen samples.

[0022] To address the above problems, the present invention also provides an electronic device, the electronic device comprising:

[0023] At least one processor; and,

[0024] A memory communicatively connected to the at least one processor; wherein,

[0025] The memory stores instructions that can be executed by the at least one processor to enable the at least one processor to perform the steps of the portable fully automated fluorescence spectroscopy sperm activity detection method described above.

[0026] This invention also protects a computer-readable storage medium storing a computer program that, when executed by a processor, implements the portable, fully automated fluorescence spectroscopy method for detecting sperm activity as described above.

[0027] This invention discloses a portable, fully automated fluorescence spectroscopy method, system, electronic device, and storage medium for detecting sperm motility. The method involves placing a semen sample into a test kit containing a sperm motility fluorescent biological staining agent, including CM-Dil bio-derived activated staining solution. Fluorescence spectral information of the semen sample is acquired using a far-field optical fluorescence microscope and a microscopic optical information acquisition component. The fluorescence spectral information is then analyzed using a spectral analysis system to obtain sperm viability, developmental status, and motility grading information. This invention offers the following advantages: it solves the problems of long testing times and high requirements for testing instruments and physician experience in existing sperm motility detection methods; with this portable, fully automated fluorescence spectroscopy method and system, individuals can complete sperm motility testing independently without going to a hospital, simply by placing a semen sample into the test kit; and the test results include sperm viability, developmental status, and motility grading information, allowing for an accurate determination of the individual's fertility. Attached Figure Description

[0028] Figure 1 This is a schematic flowchart of a portable, fully automated fluorescence spectroscopy method for detecting sperm activity according to an embodiment of the present invention.

[0029] Figure 2 This is a schematic diagram illustrating an application scenario of a portable, fully automated fluorescence spectroscopy method for detecting sperm activity according to an embodiment of the present invention.

[0030] Figure 3 This is a fluorescence intensity analysis diagram according to an embodiment of the present invention;

[0031] Figure 4 This is a statistical chart of sperm activity indicators for a portable, fully automated fluorescence spectroscopy sperm activity detection system according to an embodiment of the present invention.

[0032] Figure 5 This is a schematic diagram of a portable, fully automated fluorescence spectroscopy sperm activity detection system according to an embodiment of the present invention;

[0033] Figure 6 A schematic diagram of the internal structure of an electronic device for implementing a portable, fully automated fluorescence spectroscopy method for detecting sperm activity according to an embodiment of the present invention.

[0034] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0035] It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0036] This application's embodiments can acquire and process relevant data based on artificial intelligence (AI) and computer vision technologies. AI is the theory, methods, technologies, and application systems that use digital computers or computers-controlled machines to simulate, extend, and expand human intelligence, perceive the environment, acquire knowledge, and use that knowledge to achieve optimal results. In other words, AI is a comprehensive technology within computer science that attempts to understand the essence of intelligence and produce new intelligent machines that can react in a way similar to human intelligence. AI studies the design principles and implementation methods of various intelligent machines, enabling them to have perception, reasoning, and decision-making capabilities. AI technology is a comprehensive discipline involving a wide range of fields, encompassing both hardware and software technologies. Fundamental AI technologies generally include sensors, dedicated AI chips, cloud computing, distributed storage, big data processing, operating / interactive systems, and mechatronics. AI software technologies mainly include computer vision, speech processing, natural language processing, and machine learning / deep learning.

[0037] Specifically, as an example, Figure 1 This is a schematic flowchart illustrating a portable, fully automated fluorescence spectroscopy method for detecting sperm motility according to an embodiment of the present invention. (Refer to...) Figure 1 As shown, this invention provides a portable, fully automated method for detecting sperm motility using fluorescence spectroscopy. This method can be performed by a device, which can be implemented using software and / or hardware. It should be understood that the specific embodiments described herein are merely illustrative and are not intended to limit the scope of the invention.

[0038] A portable, fully automated method for detecting sperm motility using fluorescence spectroscopy includes steps S110–S130. S110: The semen sample to be tested is placed in a test kit containing a sperm motility fluorescent biological staining agent; the sperm motility fluorescent biological staining agent includes CM-Dil bio-derived activated staining solution. S120: The fluorescence spectral information of the semen sample to be tested is acquired using a far-field optical fluorescence microscope and a microscopic optical information acquisition component. S130: The fluorescence spectral information is analyzed using a spectral analysis system to obtain information on sperm viability, developmental status, and motility grading in the semen sample.

[0039] Figure 2This diagram illustrates an application scenario of a portable, fully automated fluorescence spectroscopy method for sperm motility detection, as provided in an embodiment of the present invention. The implementation of this portable, fully automated fluorescence spectroscopy method relies on a spectral analysis system (i.e., a server for spectral analysis), a data acquisition terminal, and a display terminal. In this embodiment, the server for spectral analysis may include, but is not limited to, a sperm motility detection model and a spectral analyzer; specifically, it can be implemented using a separate server or a server cluster composed of multiple servers. The fluorescence spectral information to be identified can be uploaded by the data acquisition terminal; for example, the data acquisition terminal sends the fluorescence spectral information of the semen sample to be tested to the server for spectral analysis. Then, the server for spectral analysis determines the sperm motility identification result for the semen sample. The sperm motility identification result for the semen sample can be pushed to the corresponding display terminal for viewing by the person being tested. The display terminal refers to an electronic device with image and text display functions, such as a smartphone, tablet computer, or computer.

[0040] In this embodiment, the spectrometer, also known as a spectrometer or optical multichannel analyzer (OMA), is widely known as a direct-reading spectrometer, a device that uses photodetectors such as photomultiplier tubes to measure the intensity of spectral lines at different wavelengths. It consists of an entrance slit, a dispersive system, an imaging system, and one or more exit slits. The dispersive element separates the electromagnetic radiation from the radiation source into the desired wavelengths or wavelength regions, and the intensity is measured at the selected wavelengths. In the embodiment described in this invention, the wavelength range is blue light (435–450 nm), green light (495–580 nm), and red light (645–760 nm). By using dispersive spectroscopy, the wavelengths are recorded sequentially on a photosensitive plate, thus presenting regular spectral lines, i.e., a spectrum. This is achieved using the dispersive and detection devices of a spectrophotometer. It conforms to the Lambert-Beer Law: A = -lg I / Io = -LgT = KCL, where I is the transmitted light intensity, Io is the emitted light intensity, T is the transmittance, and L is the optical path length through the atomizer. Since L is a constant value, A = KC. It integrates information acquisition, processing, and storage functions. Because OMA no longer uses photosensitive emulsion, it avoids and eliminates darkroom processing and subsequent cumbersome procedures, fundamentally changing traditional spectroscopic techniques, greatly improving working conditions and efficiency. Using OMA for spectral analysis offers accurate, rapid, and convenient measurements with high sensitivity, fast response time, and high spectral resolution. Measurement results can be immediately read from the display screen or output by a printer or plotter. It has been widely used in almost all spectroscopic measurements, analyses, and research, and is particularly suitable for detecting weak and transient signals.

[0041] Specifically, the acquisition terminal includes at least a far-field optical fluorescence microscope and a microscopic optical information acquisition component. The microscopic optical information acquisition component includes a CCD / CMOS integrated component. The CCD integrated component can convert light into electrical charge, store and transfer the charge, and also retrieve the stored charge to change the voltage, making it an ideal imaging element. The CCD integrated component has advantages such as small size, light weight, immunity to magnetic fields, and resistance to vibration and impact. CMOS is a complementary metal-oxide semiconductor; CMOS has N-type negatively charged and P-type positively charged semiconductors. The current generated by these two complementary effects is interpreted as an image displayed on the chip. CMOS integrated components are low-cost and energy-efficient. The far-field biofluorescence microscopy imaging used in this embodiment mainly involves labeling objects with fluorescent dyes and using a short-wavelength light source to excite the internal markers of the object, causing them to emit fluorescence, thereby improving the contrast of the object in the image. It also provides specific markers, facilitating tracking and identification in large-scale populations. Phase contrast microscopy utilizes the principle of optical interference to convert the optical path difference caused by incident light passing through a transparent object into an intensity difference, which is reflected in a CCD detector, highlighting the internal details of the transparent object and facilitating the observation of its fine internal structure. The detection light source uses an LED light source with a spectral range of 445–475 nm. The detection light source can contain multiple groups of LEDs, including blue, red, and white LEDs. Each group of LEDs is independently controlled by different switches, allowing selection of light sources within different wavelength ranges. For example, the center wavelength of the blue LED is 453 nm, suitable for exciting red fluorescent dyes / fluorescent proteins such as SRBY, Breen, I / IK, SYTO, (Cy3), and Calcein (GFP).

[0042] like Figure 2 As shown, the acquisition terminal may also include a stage for carrying the detection box, an LED light source, and a filter; furthermore, in order to further increase the accuracy of fluorescence spectral information acquisition, a fully automatic calibration unit is configured for the far-field optical fluorescence microscope. The fully automatic calibration system may include, but is not limited to, a DSP and a stepper motor for realizing automatic adjustment of the fluorescence microscope; among them, a DSP (digital signal processor) is a unique microprocessor that processes a large amount of information using digital signals.

[0043] Specifically, S110, the semen sample to be tested is placed into a test kit containing a sperm activity fluorescent biological staining agent; the sperm activity fluorescent biological staining agent includes CM-Dil biological derivative activated staining solution.

[0044] CM-Dil, a derivative of DiI, emits stable red fluorescence (Ex / Em = 553 / 570 nm) by binding to lipid molecules in membrane structures, making it suitable for monitoring cell motility and cell localization analysis. CM-Dil can freely penetrate the plasma membrane to enter cells, producing reaction products that are not membrane-permeable. At working concentrations, this dye is non-cytotoxic and does not affect cell viability or proliferation. Studies have confirmed that CM-DiI labeling results in stable intracellular fluorescence expression, with a positive labeling rate exceeding 98%, and good cell morphology.

[0045] The CM group (chloromethyl substitution group) carried by CM-Dil can react with thiol groups on peptides and proteins, thus stabilizing the molecule in aldehydes. Unlike other membrane dyes such as DiI and PKH26, some cells stained with CM-Dil cell staining agent can maintain stable labeling during subsequent fixation, permeabilization, and treatment, making it particularly suitable for simultaneous cell membrane labeling or chromatin labeling. It exhibits good dye maintenance and can be used to track cell motility. Due to its strong and stable red fluorescence (Ex / Em = 653 / 670nm), it has good fluorescence resolution with green fluorescent dyes and proteins, making it suitable for multiplex staining. The detection kit of this invention uses a non-toxic CM-Dil biological derivative. By reading the total fluorescence intensity obtained from live sperm fluorescence, sperm viability can be quickly and accurately obtained. This method of detecting sperm viability using fluorescence spectral intensity is much more accurate and faster than manual detection methods (i.e., microscopic block estimation).

[0046] As an improvement to this embodiment, when the sperm activity fluorescent biological staining agent also includes acridine orange dye, in addition to obtaining sperm sample survival rate information, developmental status information, and motility grading information, it can also obtain sperm DNA fragmentation rate information; that is, it can be applied to multi-index staining scenarios.

[0047] Specifically, acridine orange is a commonly used dye in sperm chromatin structure analysis (SCSA) to detect DNA integrity, identify subclinical sperm damage, and assess male fertility. Its characteristics are as follows: sperm with normal chromatin maintain an intact double-stranded DNA structure; double-stranded DNA fluoresces green when bound to acridine orange. Sperm with abnormal chromatin are easily denatured into single-stranded DNA; single-stranded DNA fluoresces red (715nm) when bound to acridine orange. The DNA fragmentation rate can be calculated by collecting the red and green fluorescence using a spectrometer. Acridine orange dye penetrates the cell membranes of living, necrotic, and apoptotic cells to varying degrees, exhibiting different wavelengths of fluorescence when staining chromatin. Therefore, SCSA detection can reveal the survival rate of all sperm cells (living, necrotic, and apoptotic). In summary, CM-Dil, a derivative of DiI, emits stable red fluorescence by binding to lipid molecules in membrane structures, making it suitable for monitoring cell motility and cell localization analysis. Analysis of the fluorescence spectrum can easily reveal sperm viability, developmental stage, and motility grade. Acridine orange, on the other hand, emits green fluorescence when it binds to intact DNA double strands and emits specific red light when it encounters single-stranded DNA, enabling sperm chromatin structure analysis (SCSA process). Analysis of the fluorescence spectrum can yield DNA fragmentation rate and viability. In other words, the CM-Dil bio-derived product exhibits strong and stable red fluorescence (Ex / Em = 553 / 570 nm), providing good fluorescence resolution with acridine orange green fluorescent dyes and proteins, making it suitable for multiplex staining.

[0048] Experimental Example 1

[0049] The method for obtaining sperm activity fluorescent biological staining agent, which only includes CM-Dil biological derivative activated staining solution, includes the following steps: S1111, 10 mg of CM-Dil cell staining agent is added to 10 ml of HBSS buffer and stirred to dissolve; wherein, HBSS buffer includes: 0.1 mol / L sodium chloride, 0.2 mmol / L ethylenediaminetetraacetic acid, 0.01 mol / L sodium dodecyl sarcosinate, 0.05 mol / L citric acid and 0.1 mol / L disodium hydrogen phosphate; S1112, then 250 μL of 2 mg / ml EDC and 60 μL of 2 mg / ml NHS are added, and activated at 37°C for 30 min; S1113, after the activated solution is reacted in a shaker at 37°C for 6 hours, the reaction product is purified to obtain CM-Dil biological derivative activated staining solution.

[0050] Experimental Example 2

[0051] The method for obtaining sperm activity fluorescent biological staining agent includes the following steps: S1111, 10 mg of CM-Dil cell staining agent is added to 10 ml of HBSS buffer and stirred to dissolve; wherein, the HBSS buffer includes: 0.5 mol / L sodium chloride, 2 mmol / L ethylenediaminetetraacetic acid, 0.05 mol / L sodium dodecyl sarcosinate, 0.2 mol / L citric acid and 0.5 mol / L disodium hydrogen phosphate; S1112, then 250 μL of 2 mg / ml EDC and 60 μL of 2 mg / ml NHS are added, and the mixture is activated at 37 °C for 30 min; S1113, after the activated solution is reacted in a shaker at 37 °C for 6 hours, the reaction product is purified to obtain CM-Dil biological derivative activated staining solution.

[0052] Experimental Example 3

[0053] The method for obtaining sperm activity fluorescent biological staining agent includes the following steps: S1111, 10 mg of CM-Dil cell staining agent is added to 10 ml of HBSS buffer and stirred to dissolve; wherein, HBSS buffer includes: 0.3 mol / L sodium chloride, 1.7 mmol / L ethylenediaminetetraacetic acid, 0.03 mol / L sodium dodecyl sarcosinate, 0.15 mol / L citric acid and 0.3 mol / L disodium hydrogen phosphate. S1112. Then add 250 μL of 2 mg / mL EDC and 60 μL of 2 mg / mL NHS, and activate at 37°C for 30 min. S1113. Add 40 mg of acridine orange dye to 10 mL of HBSS buffer, stir to dissolve, and obtain an acridine orange dye mixed solution. The HBSS buffer includes: 0.3 mol / L sodium chloride, 1.7 mmol / L EDTA, 0.03 mol / L sodium dodecyl sarcosinate, 0.15 mol / L citric acid, and 0.3 mol / L disodium hydrogen phosphate. S1114. Add the activated CM-Dil bio-derived staining solution to the acridine orange dye mixed solution, and react in a shaker at 37°C for 3 hours. After purification, obtain a complex solution of sperm-active fluorescent biological staining agent. S1115. Add DMF and DMSO stock solution to prepare a 2 mmol / L concentration of sperm-active fluorescent biological staining agent.

[0054] Experiment Example 4

[0055] The method for obtaining sperm activity fluorescent biological staining agent includes the following steps: S1111, 10 mg of CM-Dil cell staining agent is added to 10 ml of HBSS buffer and stirred to dissolve; wherein, HBSS buffer includes: 0.4 mol / L sodium chloride, 0.8 mmol / L ethylenediaminetetraacetic acid, 0.02 mol / L sodium dodecyl sarcosinate, 0.09 mol / L citric acid and 0.2 mol / L disodium hydrogen phosphate. S1112. Then add 250 μL of 2 mg / mL EDC and 60 μL of 2 mg / mL NHS, and activate at 37°C for 30 min. S1113. Add 40 mg of acridine orange dye to 10 mL of HBSS buffer, stir to dissolve, and obtain an acridine orange dye mixed solution. The HBSS buffer includes: 0.4 mol / L sodium chloride, 0.8 mmol / L EDTA, 0.02 mol / L sodium dodecyl sarcosinate, 0.09 mol / L citric acid, and 0.2 mol / L disodium hydrogen phosphate. S1114. Add the activated CM-Dil bio-derived staining solution to the acridine orange dye mixed solution, and react in a shaker at 37°C for 3 hours. After purification, obtain a complex solution of sperm-active fluorescent biological staining agent. S1115. Add DMF and DMSO stock solution to prepare a 1 mmol / L concentration of sperm-active fluorescent biological staining agent.

[0056] Experimental Example 5

[0057] The method for obtaining sperm activity fluorescent biological staining agent includes the following steps: S1111, 10 mg of CM-Dil cell staining agent is added to 10 ml of HBSS buffer and stirred to dissolve; wherein, HBSS buffer includes: 0.1 mol / L sodium chloride, 0.2 mmol / L ethylenediaminetetraacetic acid, 0.02 mol / L sodium dodecyl sarcosinate, 0.06 mol / L citric acid and 0.4 mol / L disodium hydrogen phosphate. S1112. Then add 250 μL of 2 mg / mL EDC and 60 μL of 2 mg / mL NHS, and activate at 37°C for 30 min. S1113. Add 40 mg of acridine orange dye to 10 mL of HBSS buffer, stir to dissolve, and obtain an acridine orange dye mixed solution. The HBSS buffer includes: 0.1 mol / L sodium chloride, 0.2 mmol / L ethylenediaminetetraacetic acid, 0.02 mol / L sodium dodecyl sarcosinate, 0.06 mol / L citric acid, and 0.4 mol / L disodium hydrogen phosphate. S1114. Add the activated CM-Dil biological derivative staining solution to the acridine orange dye mixed solution, and react in a shaker at 37°C for 3 hours. After purification, obtain a complex solution of sperm activity fluorescent biological staining agent. S1115. Add DMF and DMSO stock solution to prepare a 1.5 mmol / L concentration of sperm activity fluorescent biological staining agent.

[0058] Dimethylformamide (DMF) is a transparent liquid miscible with water and most organic solvents. DMSO is dimethyl sulfoxide (DMSO), a colorless liquid and an important polar aprotic solvent miscible with many organic solvents and water. EDC is an aqueous carbodiimide, mainly used to activate carboxyl groups, enabling carboxyl, amino, and hydroxyl groups in aqueous systems to undergo coupling reactions at room temperature or even low temperatures. NHS is a chemical substance of N-hydroxythiosuccinimide. Thermo Scientific Pierce NHS is a chemical modifying agent that converts carboxyl groups into amine-reactive NHS esters, used in bio-coupling, cross-linking, labeling, and immobilization methods. It can control and modify carbodiimide cross-linking reactions involving carboxylates (-COOH) coupled to primary amines (-NH2). Where specific techniques or conditions are not specified in the examples, they should be performed according to the techniques or conditions described in the literature in this field, or according to the product instructions. Reagents or instruments whose manufacturers are not specified can be purchased from reputable suppliers.

[0059] In summary, this invention discloses a staining agent for sperm motility detection, namely CM-Dil bio-derived activated staining solution. This invention also discloses a detection kit for sperm motility detection, the kit containing CM-Dil bio-derived activated staining solution, or a sperm motility fluorescent biological staining agent comprising CM-Dil bio-derived activated staining solution and acridine orange dye.

[0060] The semen samples to be tested were placed in the test kit, and the sperm activity fluorescent biological staining agents prepared using the methods described in Experiments 1 and 2 were added for staining. Then, the fluorescence spectrum information of the semen samples was acquired using a far-field optical fluorescence microscope and a microscopic optical information acquisition component. Analysis of the fluorescence spectra easily revealed the sperm viability, developmental stage, and motility grade. The sperm activity fluorescent biological staining agents prepared using the methods described in Experiments 3-5 were also added for staining. The fluorescence spectrum information of the semen samples was then acquired using a far-field optical fluorescence microscope and a microscopic optical information acquisition component. Analysis of the fluorescence spectra easily revealed the sperm viability, DNA fragmentation rate, developmental stage, and motility grade.

[0061] This invention utilizes a non-toxic Dil derivative, which labels cells by binding to lipid molecules in the membrane structure and exhibits strong and stable red fluorescence, making it suitable for quantitative cell analysis. Therefore, by reading the total intensity of the specific fluorescence (green 515nm) emitted by live sperm, sperm viability can be obtained quickly and accurately. This is a statistical calculation method based on fluorescence peaks, which is significantly more accurate and faster than estimation methods using microscopic block detection.

[0062] S120. Obtain the fluorescence spectrum information of the semen sample to be tested through a far-field optical fluorescence microscope and a microscopic optical information acquisition component.

[0063] In practical implementation, the detection light source configured for the far-field optical fluorescence microscope and the microscopic optical information acquisition component uses an LED light source with a spectral range of 445–475 nm. The detection light source can contain multiple groups of LEDs, including blue, red, and white LEDs. Each group of LEDs is independently controlled by different switches, allowing selection of light sources with different wavelength ranges. For example, the center wavelength of a blue LED is 453 nm, suitable for exciting red fluorescent dyes / fluorescent proteins such as SRBY, Breen, I / IK, SYTO, (Cy3), and Calcein (GFP). Before detecting analytes containing the aforementioned fluorescent dyes / fluorescent proteins, the models of the detection light source, detection filter, and filter element need to be determined according to the fluorescent dye / fluorescent protein or other fluorescent substance. Specifically, for red fluorescent substances, the transmission spectral range of the detection filter is 325 nm–500 nm, and the transmission spectral range of the filter element is 500 nm–2500 nm. For example, if the detection light source is blue, with a wavelength of 476–495 nm, the corresponding filter model can be JB470 (Xtj470±10nm); if the detection light source is green, with a wavelength of 495–570 nm, the corresponding filter model can be a golden yellow cutoff glass (JB510, Xtj510±10nm) or JB490 (Xtj490±10nm), which can filter out stray light with wavelengths below 500 nm and can be used with a blue LED to detect green fluorescent dyes / fluorescent proteins; if the detection light source is yellow, with a wavelength of 570–590 nm, the corresponding filter model can be CB565 (Xtj565±10nm) or CB580 (Xtj580±10nm); and for red light with a wavelength of 620–750 nm, the corresponding filter model can be HB610 / 620 / 630.

[0064] S130. The fluorescence spectral information is analyzed by a spectral analysis system to obtain information on sperm survival rate, developmental status, and motility grading of the semen sample.

[0065] The spectrometer separates the electromagnetic radiation from the radiation source into the desired wavelength region using a dispersive element and measures the intensity at the selected wavelength. The wavelength regions include: 435–450 nm for blue light; 495–580 nm for green light; and 645–760 nm for red light.

[0066] In one specific embodiment, the method for obtaining sperm survival rate information, developmental status information, and motility grading information of semen samples by performing data analysis on fluorescence spectral information through a spectral analysis system includes: preprocessing the fluorescence spectral information using a preset sperm motility detection model to obtain denoised fluorescence spectral information; and performing data analysis on the denoised fluorescence spectral information using a spectral analyzer to obtain sperm survival rate information, density information, and motility grading information of semen samples.

[0067] In other words, to further improve the accuracy of sperm motility detection and reduce substrate fluorescence interference (blue 435-450nm) and pattern fluorescence loss (red and green), this invention introduces a simulation mapping optimization model. The input to the generative model is the collected fluorescence spectral information, which is mapped to a new data space (without background noise) through the generator's internal network structure. The resulting probability distribution model G(z) reaches equilibrium in the dynamic struggle between the optimal generator and the optimal discriminator, thus obtaining the optimal generative model, i.e., the sperm motility detection model. The objective function of the sperm motility detection model is as follows:

[0068]

[0069] In this design, G represents the generator, and D represents the discriminator. The discriminator's judgment result is fed back to the generator, which then adjusts its structure based on the result to obtain a better generator structure. Simultaneously, the generator sends the optimized probability distribution model to the discriminator for processing. The discriminator optimizes its own parameters to maximize the distinction between the simulated and real distributions. The entire process iterates continuously until a dynamic equilibrium is reached, achieving the optimal structure for both the discriminator and the generator. Ultimately, a simulated distribution similar to the real distribution is generated. Specifically, this invention receives sperm density and sperm motility information in the form of fluorescence spectral information, performs noise reduction processing, and provides a more accurate raw basis for further digital processing and automatic conversion into corresponding sperm motility quantification data.

[0070] It should be noted that during spectral analysis, sperm viability can be quickly and accurately obtained by reading the total fluorescence intensity of live sperm fluorescence (red fluorescence of the Dil derivative, Ex / Em = 653 / 670nm). DNA fragmentation rate can be calculated by separately reading the fluorescence intensity of the collected red fluorescence (715nm) and green fluorescence.

[0071] After analyzing the fluorescence spectral information through a spectral analysis system to obtain information on sperm survival rate, developmental status, and motility grading in the semen sample, the data is displayed on a display terminal.

[0072] The data processing module in a spectral analysis system may include a local data processing module and a remote data processing module. The local data processing module is operatively coupled, for example, via a wired or wireless communication link, to remote processing modules and a remote data repository (the Internet), such that these remote modules are operatively coupled to each other and can be used as resources for the local processing and data modules. In one embodiment, the remote data repository may include a relatively large-scale digital data storage facility, accessible via the Internet or other network configurations in a "cloud" resource configuration. In one embodiment, all data is stored and all computations are performed in the local processing and data modules, allowing for fully autonomous use from any remote module. Specifically, the data processing module can manage data from sperm quality analysis, report test results, patient information, and quality control settings; it can be used to set the controls and system defaults of the analysis system, and can adopt different versions of standards according to the user's laboratory practices. In addition, on the PC screen serving as the display terminal, the subject can observe the semen sample "in real time," enabling video signal reproduction, screenshotting, and video capture. Furthermore, the spectral analysis system plots a standard curve of fluorescence values ​​against sample concentration, allowing direct observation of the different discrete spectral information reflected by sperm with varying motility. Utilizing advanced spectral analysis technology, the spectral information can be digitally converted to obtain a motility grading histogram for assessing sperm activity. This can include, but is not limited to, sperm motility grading histograms, sperm concentration histograms, sperm motility statistical histograms, and sperm motility statistical histograms. Each type of graph displays specific motility data for four sperm types: progressively motile sperm, non-progressively motile sperm, hyperactive sperm, and immotile sperm.

[0073] Changes in phospholipids, fatty acids, and sulfhydryl groups in the sperm membrane structure during sperm maturation signify alterations in the sperm's developmental state. The composition of lipid molecules differs at different maturation stages. Therefore, by using CM-DIL fluorescent dye, which binds to lipid molecules on the membrane structure, and observing the specific spectra emitted at different developmental stages, the developmental state of sperm can be monitored. Quantitative cellular analysis can provide information on the number and proportion of sperm at various maturation stages within the field of view. Sperm motility in the epididymis undergoes regular changes. Based on these specific changes, sperm maturation can be divided into six stages: initial stationary oscillation, followed by circular motion, then full maturity and spiral forward movement. After this, sperm lose motility after a period of flaccidity. Therefore, observing sperm motility is an indicator of sperm maturity, and sperm motility is classified into four grades (A, B, C, D) according to WHO standards. Grade A indicates excellent sperm motility, most importantly, rapid linear movement, meaning it can fertilize an egg very quickly; Grade B indicates decent sperm motility, capable of forward movement, but at a moderate speed; Grade C indicates moderate sperm motility, exhibiting only curvilinear movement, but still capable of fertilization; Grade D indicates very poor sperm motility, almost only exhibiting stationary movement, or even lacking any motility. If the number of grade C or D sperm in semen exceeds 50%, and the number of grade A sperm is less than 25%, it is considered low sperm motility.

[0074] In one specific embodiment, the portable fully automated fluorescence spectroscopy sperm motility detection system of the present invention is placed in a hospital reproductive laboratory or reproductive testing center. After collecting their own semen sample, the individual places the sample in a disposable collection cup. The sample is then placed in the test kit, i.e., CM-Dil derivative is added for staining, and allowed to stand for half a minute to allow for full reaction. The semen sample is placed on the sample holder of the stage at the collection terminal, and information is acquired using a microscopic imaging system. Based on the fluorescence intensity and discrete spectral information, sperm viability and sperm motility grading indicators are obtained. By comparing with reference information, the individual's sperm motility status can be determined. After the test is completed, the test results are displayed on the display terminal, and a test report can be generated. A standard curve is plotted between the fluorescence values ​​obtained by the analysis unit and the sample concentration to obtain a linear regression equation, allowing direct observation of the different discrete spectral information reflected by sperm with different motility levels.

[0075] Figure 3A fluorescence intensity analysis diagram from one embodiment is presented. The electromagnetic radiation from the radiation source is separated into the desired wavelengths or wavelength regions using a dispersive element, and the intensity is measured at the selected wavelengths. By using dispersive spectroscopy, the wavelengths are recorded sequentially on a photosensitive plate, resulting in regular spectral lines. In this embodiment, the blue light in the wavelength range of 435–450 nm is caused by substrate fluorescence interference from the buffer solution, and the green light in the wavelength range of 495–580 nm is the fluorescence emitted by acridine orange staining during the SCSA process. The discrete spectral information includes information such as DNA fragmentation rate and viability. This fluorescence information is suitable for quantitative cell analysis; therefore, by reading the total intensity of the specific fluorescence (green 515 nm) emitted by live sperm, sperm viability can be obtained quickly and accurately. In summary, this is a statistical calculation method based on fluorescence peaks. Compared to estimation methods using microscopic block detection, this detection method utilizing fluorescence spectroscopy is significantly more accurate and faster.

[0076] The red light in the wavelength range of 645–760 nm is mainly emitted by the combination of cm-dil biological dyes with certain molecules such as lipids, resulting in stable red fluorescence. It is suitable for monitoring cell movement and cell localization analysis. By analyzing the fluorescence spectrum, the developmental status and motility grade of sperm can be easily obtained.

[0077] Furthermore, Figure 4 This is a statistical chart of sperm activity indicators from a portable, fully automated fluorescence spectroscopy sperm activity detection system according to an embodiment of the present invention. A is a histogram of sperm motility grading (unquantified), B is a histogram of sperm concentration, C is a histogram of sperm motility statistics, and D is a histogram of sperm motility statistics; PR represents progressively motile sperm, NP represents non-progressively motile sperm, SPR represents highly activated sperm, and IM represents immotile sperm. Figure 5 As shown in the histogram, based on the grading of the parameters, and according to the WHO Fifth Edition Manual of Semen Analysis, the total sperm motility (PR+NP) of this semen sample is ≥40%, progressive motility is ≥32%, sperm concentration is ≥25%, and sperm motility is graded as Level 1.

[0078] This invention utilizes advanced spectral analysis technology to directly observe the different discrete spectral information reflected by sperm with varying motility. Therefore, histograms grading sperm motility can be obtained based on this spectral information. This allows for quantitative analysis of live sperm cells and monitoring of sperm cell motility characteristics, offering high efficiency, objectivity, and precision. Furthermore, histograms of various sperm motility parameters, speed, and motility grading provide a basis for assessing sperm activity.

[0079] In summary, this invention presents a portable, fully automated fluorescence spectroscopy method for sperm motility detection. It combines non-cytotoxic biological derivative fluorescent dyes with modern computer technology and advanced spectral analysis techniques, resulting in a fluorescence spectroscopy sperm motility detection system. This system is characterized by high efficiency, objectivity, and high precision. Furthermore, histograms of various sperm motility parameters, velocity, and motility grading provide a basis for sperm motility assessment. Primarily applicable to home testing or community hospitals, this method combines testing reagents with the instrument to perform sperm motility detection, offering a more convenient, faster, and more efficient testing service for those preparing for pregnancy.

[0080] like Figure 5 As shown, the portable fully automated fluorescence spectroscopy sperm motility detection system 500 provided by the present invention can be installed in an electronic device. Depending on the functions implemented, the portable fully automated fluorescence spectroscopy sperm motility detection system 500 may include a detection unit 510, a spectral information acquisition unit 520, and a data analysis unit 530.

[0081] In this embodiment, the functions of each module / unit are as follows: the detection unit 510 is used to put the semen sample to be tested into a detection kit containing a sperm activity fluorescent biological staining agent; the sperm activity fluorescent biological staining agent includes CM-Dil biological derivative activation staining solution.

[0082] The spectral information acquisition unit 520 is used to acquire the fluorescence spectral information of the semen sample to be tested through a far-field optical fluorescence microscope and a microscopic optical information acquisition component.

[0083] The data analysis unit 530 is used to analyze fluorescence spectral information through a spectral analysis system to obtain sperm survival rate, developmental status, and motility grading information from semen samples. The unit described in this invention can also be called a module, referring to a series of computer program segments that can be executed by the processor of an electronic device and perform a specific function, stored in the memory of the electronic device.

[0084] More specific implementations of the portable fully automated fluorescence spectroscopy sperm motility detection system provided by the present invention can all refer to the above-described embodiments of the portable fully automated fluorescence spectroscopy sperm motility detection method, and will not be listed one by one here.

[0085] The portable, fully automated fluorescence spectroscopy sperm motility detection system provided by this invention combines non-cytotoxic biological derivative fluorescent dyes with modern computer technology and advanced spectral analysis techniques. This results in a fluorescence spectroscopy sperm motility detection system that is highly efficient, objective, and accurate. Furthermore, histograms of various sperm motility parameters, velocity, and motility grading provide a basis for sperm motility assessment. It is primarily applied in home testing or community hospital settings, using a combination of testing reagents and instruments to perform sperm motility detection, providing a more convenient, faster, and more efficient testing service for people preparing for pregnancy.

[0086] like Figure 6 As shown, the present invention provides an electronic device 6 for a portable, fully automated fluorescence spectroscopy method for detecting sperm activity.

[0087] The electronic device 6 may include a processor 60, a memory 61 and a bus, and may also include a computer program stored in the memory 61 and executable on the processor 60, such as a sperm motility detection program 62.

[0088] The memory 61 includes at least one type of readable storage medium, such as flash memory, portable hard drive, multimedia card, card-type memory (e.g., SD or DX memory), magnetic memory, magnetic disk, optical disk, etc. In some embodiments, the memory 61 can be an internal storage unit of the electronic device 6, such as the portable hard drive of the electronic device 6. In other embodiments, the memory 61 can be an external storage device of the electronic device 6, such as a plug-in portable hard drive, smart media card (SMC), secure digital card (SD), flash card, etc., equipped on the electronic device 6. Furthermore, the memory 61 can include both internal and external storage units of the electronic device 6. The memory 61 can be used not only to store application software and various types of data installed on the electronic device 6, such as the code of a sperm activity detection program, but also to temporarily store data that has been output or will be output.

[0089] In some embodiments, the processor 60 may be composed of integrated circuits, such as a single packaged integrated circuit or multiple integrated circuits with the same or different functions, including combinations of one or more central processing units (CPUs), microprocessors, digital processing chips, graphics processors, and various control chips. The processor 60 is the control unit of the electronic device, connecting various components of the entire electronic device through various interfaces and lines. It executes programs or modules (such as sperm motility detection programs) stored in the memory 61, and calls data stored in the memory 61 to perform various functions of the electronic device 6 and process data.

[0090] The bus can be a Peripheral Component Interconnect (PCI) bus or an Extended Industry Standard Architecture (EISA) bus, etc. This bus can be divided into an address bus, a data bus, a control bus, etc. The bus is configured to enable communication between the memory 61 and at least one processor 60, etc.

[0091] Figure 6 Only electronic devices with components are shown; it will be understood by those skilled in the art that... Figure 6 The structure shown does not constitute a limitation on the electronic device 6, and may include fewer or more components than shown, or combine certain components, or have different component arrangements.

[0092] For example, although not shown, the electronic device 6 may also include a power supply (such as a battery) to power the various components. Preferably, the power supply can be logically connected to the at least one processor 60 through a power management device, thereby enabling functions such as charging management, discharging management, and power consumption management. The power supply may also include one or more DC or AC power supplies, recharging devices, power fault detection circuits, power converters or inverters, power status indicators, and other arbitrary components. The electronic device 6 may also include various sensors, Bluetooth modules, Wi-Fi modules, etc., which will not be described in detail here.

[0093] Furthermore, the electronic device 6 may also include a network interface. Optionally, the network interface may include a wired interface and / or a wireless interface (such as a Wi-Fi interface, a Bluetooth interface, etc.), which is typically used to establish communication connections between the electronic device 6 and other electronic devices.

[0094] Optionally, the electronic device 6 may further include a user interface, which may be a display, an input unit (such as a keyboard), or a standard wired or wireless interface. Optionally, in some embodiments, the display may be an LED display, a liquid crystal display, a touch-sensitive liquid crystal display, or an OLED (Organic Light-Emitting Diode) touchscreen. The display may also be appropriately referred to as a screen or display unit, used to display information processed in the electronic device 6 and to display a visual user interface.

[0095] It should be understood that the embodiments described are for illustrative purposes only and are not limited to this structure in the scope of the patent application.

[0096] The sperm activity detection program 62 stored in the memory 61 of the electronic device 6 is a combination of multiple instructions. When run in the processor 60, it can achieve the following: placing the semen sample to be tested into a test kit containing a sperm activity fluorescent biological staining agent; the sperm activity fluorescent biological staining agent includes CM-Dil biological derivative activation staining solution; acquiring the fluorescence spectrum information of the semen sample to be tested through a far-field optical fluorescence microscope and a microscopic optical information acquisition component; and performing data analysis on the fluorescence spectrum information through a spectral analysis system to obtain sperm survival rate information, developmental status information, and motility grading information of the semen sample.

[0097] Specifically, the processor 60's implementation method for the above instructions can be found in [reference needed]. Figure 1 The descriptions of the relevant steps in the corresponding embodiments are not repeated here. It should be emphasized that, to further ensure the privacy and security of the above sperm motility detection procedure, the sperm motility detection procedure is stored in the nodes of the blockchain on which this server cluster is located.

[0098] Furthermore, if the modules / units integrated in the electronic device 6 are implemented as software functional units and sold or used as independent products, they can be stored in a computer-readable storage medium. The computer-readable medium may include: any entity or device capable of carrying the computer program code, a recording medium, a USB flash drive, a portable hard drive, a magnetic disk, an optical disk, a computer memory, or a read-only memory (ROM).

[0099] This invention also provides a computer-readable storage medium, which may be non-volatile or volatile. The storage medium stores a computer program, which, when executed by a processor, performs the following: placing a semen sample to be tested into a test kit containing a sperm activity fluorescent biological staining agent; the sperm activity fluorescent biological staining agent includes CM-Dil biological derivative activated staining solution; acquiring the fluorescence spectrum information of the semen sample to be tested using a far-field optical fluorescence microscope and a microscopic optical information acquisition component; and performing data analysis on the fluorescence spectrum information using a spectral analysis system to obtain sperm survival rate information, developmental status information, and motility grading information of the semen sample.

[0100] Specifically, the specific implementation method of the computer program when executed by the processor can be referred to the description of the relevant steps in the embodiment of a portable fully automated fluorescence spectroscopy sperm activity detection method, which will not be repeated here.

[0101] In the several embodiments provided by this invention, it should be understood that the disclosed devices, apparatuses, and methods can be implemented in other ways. For example, the apparatus embodiments described above are merely illustrative; for instance, the division of modules is only a logical functional division, and other division methods may be used in actual implementation.

[0102] The modules described as separate components may or may not be physically separate. The components shown as modules may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the modules can be selected to achieve the purpose of this embodiment according to actual needs.

[0103] Furthermore, the functional modules in the various embodiments of the present invention can be integrated into one processing unit, or each unit can exist physically separately, or two or more units can be integrated into one unit. The integrated unit can be implemented in hardware or in the form of hardware plus software functional modules.

[0104] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention.

[0105] Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be embraced within the invention. No appended diagram markings in the claims should be construed as limiting the scope of the claims.

[0106] The blockchain referred to in this invention is a novel application model of computer technologies such as distributed data storage, peer-to-peer transmission, consensus mechanisms, and encryption algorithms. Essentially, a blockchain is a decentralized database, a chain of data blocks linked together using cryptographic methods. Each data block contains information about a batch of network transactions, used to verify the validity of the information (anti-counterfeiting) and generate the next block. A blockchain can include an underlying blockchain platform, a platform product service layer, and an application service layer.

[0107] Furthermore, it is clear that the word "comprising" does not exclude other units or steps, and the singular does not exclude the plural. Multiple units or devices recited in a system claim may also be implemented by a single unit or device through software or hardware. The term "second class" is used to indicate names and does not indicate any specific order.

[0108] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A portable full-automatic fluorescent spectrum sperm activity detection method, characterized in that, The method comprises: Placing the semen sample to be detected into a detection kit added with a sperm activity fluorescent biological dye; the sperm activity fluorescent biological dye comprises a CM-Dil biological derivative activated dye solution; Obtaining fluorescence spectrum information of the semen sample to be detected through a far-field optical fluorescence microscope and a microscopic optical information acquisition component; The method for obtaining survival rate information, development state information and activity classification information of sperm in the semen sample through data analysis of the fluorescence spectrum information by a spectrum analysis system comprises: Preprocessing the fluorescence spectrum information by using a preset sperm activity detection model to obtain denoised fluorescence spectrum information; Obtaining survival rate information, development state information and activity classification information of sperm in the semen sample through data analysis of the denoised fluorescence spectrum information by a spectrum analyzer; The method for obtaining the CM-Dil biological derivative activated dye solution comprises: Adding 10 mg of CM-Dil cell dye into 10 ml of HBSS buffer solution and stirring to dissolve; Then adding 250 ul of EDC with a concentration of 2 mg / ml and 60 ul of NHS with a concentration of 2 mg / ml, and activating at 37 DEG C for 30 min; After the activated solution is subjected to a shaking bed reaction at 37 DEG C for 6 hours, the reaction product is purified to obtain the CM-Dil biological derivative activated dye solution.

2. The portable full-automatic fluorescent spectrum sperm activity detection method according to claim 1, wherein, The HBSS buffer includes: 0.1 0.5 mol / L sodium chloride, 0.2 2 mmol / L ethylenediaminetetraacetic acid, 0.01 0.05 mol / L sodium dodecylsarcosinate, 0.05 0.2 mol / L citric acid, and 0.1 0.5 mol / L disodium hydrogen phosphate.

3. A portable fully automated fluorescent spectroscopy based sperm motility detection method as claimed in claim 1, wherein, The spectrum analyzer separates electromagnetic radiation of a radiation source into a required wavelength region through a dispersion element and performs intensity measurement at a selected wavelength; the wavelength region comprises: a wavelength region of blue light is 435-450 nm; a wavelength region of green light is 495-580 nm; and a wavelength region of red light is 645-760 nm.

4. The portable full-automatic fluorescent spectrum sperm activity detection method according to claim 1, wherein, When the sperm activity fluorescent biological dye further comprises acridine orange dye, in addition to obtaining survival rate information, development state information and activity classification information of the semen sample, DNA fragmentation rate information of sperm can also be obtained.

5. A portable fully automatic fluorescent spectrum sperm activity detection system, characterized in that, The method comprises: A detection unit for placing the semen sample to be detected into a detection kit added with a sperm activity fluorescent biological dye; the sperm activity fluorescent biological dye comprises a CM-Dil biological derivative activated dye solution; A spectrum information acquisition unit for obtaining fluorescence spectrum information of the semen sample to be detected through a far-field optical fluorescence microscope and a microscopic optical information acquisition component; A data analysis unit for obtaining survival rate information, development state information and activity classification information of sperm in the semen sample through data analysis of the fluorescence spectrum information by a spectrum analysis system.

6. An electronic device, comprising: The electronic device comprises: At least one processor; and A memory connected in communication with the at least one processor; wherein The memory stores instructions executable by the at least one processor, and the instructions are executed by the at least one processor to enable the at least one processor to perform steps in a portable full-automatic fluorescence spectrum sperm activity detection method according to any one of claims 1 to 4.

7. A computer readable storage medium storing a computer program, characterized in that, The computer program is executed by a processor to implement a portable full-automatic fluorescence spectrum sperm activity detection method according to any one of claims 1 to 4.

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