WO2020133635A1

WO2020133635A1 - Fully automatic sperm cell detector

Info

Publication number: WO2020133635A1
Application number: PCT/CN2019/073863
Authority: WO
Inventors: 李翼飞; 李生杰; 汪谢华; 柴艳华
Original assignee: 深圳天依生命健康科技有限公司
Priority date: 2018-12-26
Filing date: 2019-01-30
Publication date: 2020-07-02
Also published as: CN109612911A; CN109612911B

Abstract

A fully automatic sperm cell detector, comprising a detection platform (10), a control system (60) provided on the detection platform (10), and a sample processing system (20), liquid flow system (30), laser optical system (40) and photoelectric detection system (50) that are all connected to the control system (60). The sample processing system (20) is used to count sperm cells in a semen sample to be processed, and perform a dyeing reaction treatment on the sperm cells in the semen sample to obtain a dyed sperm sample. The liquid flow system (30) is connected to the sample processing system (20), and is used to carry out mixed reaction, flowing, detection and recovery processing of the dyed sperm sample outputted from the sample processing system (20) and sheath fluid. The control system (60) is used to control the sample processing system (20), the liquid flow system (30), the laser optical system (40) and the photoelectric detection system (50), and then conduct fully automatic detection and analysis on the semen sample and generate an analysis result. The present detector implements the fully automatic high-throughput rapid detection and analysis of semen samples. A user need only place the semen samples in a sample pool to carry out a fully automatic operation and automatically output the detection and analysis results.

Description

Automatic sperm cell detector

Technical field

The invention relates to the technical field of new medical grade clinical detection, in particular to a fully automatic sperm cell detector.

Background technique

Sperm quality evaluation is the core key for diagnosing male (male) fertility. Test items include sperm density, activity rate, deformity rate, nuclear integrity, mitochondrial membrane potential, acrosome reaction and other individual or group indicators. At present, the main detection equipment includes microscope, fluorescence microscope, general-purpose flow cytometer, etc. The main detection methods include microscope artificial microscopy, computer-assisted sperm analysis system (CASA) auxiliary analysis and flow cytometry computer equipped with a cell analysis system standard Assist manual analysis. The method of universal flow cytometry detection and analysis has begun to be applied to the detection of clinical indicators of sperm. Flow cytometry has great technical advantages in rapid and large-scale detection of cells, which can achieve accurate detection of high-throughput samples, and Has expanded the detectable index parameters of clinical sperm quality, and has been widely concerned by clinicians. However, the flow cytometer is mainly imported equipment, which is expensive, and there is currently no instrument dedicated to sperm cell detection in the world. In actual use, long-term training is required, and during the detection process, manual voltage adjustment and compensation are required. , Flow rate and other indicators, setting each relevant area or door, it is difficult to meet the objective, repeatability and reference of clinical testing requirements. After the test is completed, it is necessary to manually distinguish the areas and doors when analyzing the results, which is difficult to meet Day-to-day repeatability and room-to-room reference required for clinical testing. A Flow cytometers are mostly universal detection platforms or multi-laser multi-channel multi-parameter instruments for scientific research. In specific application scenarios, sample processing is usually separated from detection, and there is no automatic sample processing, reaction staining and sampling in the world. The instrument that is integrated with detection and automation, the general clinical flow cytometer is more suitable for scientific research, or the use of a small number of blood cell indicators with high detection requirements.

Microscope artificial microscopy is currently the most mainstream application technology. It mainly uses artificial microscopy to count, distinguish viability and morphological observation with a microscope after preparing a seminal cell smear. There is also a computer-aided system to assist in analyzing the images obtained by the microscope. The main disadvantage is that it requires a lot of manpower. If a computer-aided analysis system is added, the detection cost will also increase significantly. The most important thing is that only a small number of indicators such as sperm quantity, vitality, and morphology can be detected. Other important indicators such as sperm cell nuclear integrity and mitochondrial function , Acrosomal membrane and other indicators need to be prepared after cell pictures are labeled with fluorescent probes, and then analyzed under a microscope using a fluorescent microscope, and there is currently no clinically-analyzed automatic analysis assistance software, which is costly and requires a lot of manpower for artificial naked eyes Observe and judge and count.

technical problem

At present, there is a lack of fully-automatic integrated high-throughput and low-cost detection instruments dedicated to the analysis of sperm cells in clinical semen testing at home and abroad. The sample processing, instrument testing, and result analysis brought by the existing instruments must all be carried out separately and required Manual operation is manual, which results in complicated detection operation and cumbersome manual sample processing steps, which requires special personnel training for equipment maintenance and operation problems.

Technical solution

The technical problem to be solved by the present invention is to provide a fully automatic sperm cell detector in view of the above-mentioned defects of the prior art. The technical solution adopted by the present invention to solve its technical problems is that, according to an aspect of the present invention, a fully automatic sperm cell detector is provided, which includes a test bench, a control system provided on the test bench, and are connected to the control system Sample processing system, liquid flow system, laser optical system and photoelectric detection system; specifically, the sample processing system is used for counting statistics of sperm cells in the semen sample to be processed, and for staining sperm cells in the semen sample A dyed sperm sample is obtained by reaction processing; the flow system is connected to a sample processing system for mixing reaction, flow, detection and recovery processing of the dyed sperm sample and sheath fluid output by the sample processing system; the control system is used for Control the sample processing system, liquid flow system, laser optical system and photoelectric detection system, and then carry out automatic detection and analysis of the semen samples and generate analysis results.

Preferably, the sample processing system includes a semen sample pool, a sample counting device, a sample injection needle, a sheath liquid pool, a cleaning liquid pool, a reaction staining device, multiple transfer tubes, multiple solenoid valves, and a first sheath liquid peristaltic pump , A first buffer filtration device, a sample injection pump, a staining reaction cell, and an automatic temperature control device; the semen sample pool is connected to the staining reaction cell in sequence through a sample counting device, a sample injection needle, a solenoid valve, and a sample injection pump, and the semen sample The pool is used to store the semen sample to be tested, and the semen sample includes a large number of sperm cells; the sample counting device is connected to the semen sample pool and the sample injection needle, and is used for each sperm cell in the semen sample passing therethrough Counting and size measurement; the sheath liquid pool is used to store the sheath liquid to be mixed, and the cleaning liquid pool is used to store the cleaning liquid; the sheath liquid pool and the cleaning liquid pool are connected to the reaction dyeing device through the transfer tube, the sheath liquid The sheath liquid in the pool and the cleaning liquid in the cleaning liquid pool are used for cleaning the reaction dyeing device and the liquid flow system after the completion of the dyeing reaction;

The reaction dyeing device is connected to the dyeing reaction tank through an electromagnetic valve and a first sheath liquid peristaltic pump. The first buffer filter device is used for pulse filtering and impurity filtering of the sheath liquid output by the sheath liquid pool; the reaction dyeing device is used for Provide at least one reaction reagent and/or staining reagent;

The automatic temperature control device is arranged below the dyeing reaction tank and is used to heat the dyeing reaction pool according to the reaction temperature requirements; the reaction reagent and/or the dyeing reagent in the reaction dyeing device are sucked out by the first sheath liquid peristaltic pump and enter Dyeing reaction pool; the semen sample in the semen sample pool enters the dyeing reaction pool through the sample injection pump, and the semen sample reacts with at least one reaction reagent and/or dyeing reagent in the dyeing reaction pool to form a dyed sperm sample.

Preferably, the sample counting device includes a negative electrode, a positive electrode, and a voltage pulse measurement device, and the voltage pulse measurement device is electrically connected to the negative electrode and the positive electrode; the negative electrode and the positive electrode together form a sperm cell through Micropore channel, the micropore channel is set on one side of the semen sample pool and on the other side of the sample injection needle; when different sperm cells in the semen sample pass through the micropore channel, different voltage pulse signals are generated. The measuring device will measure different voltage pulse signals. The voltage pulse signal includes information on the number and size of sperm cells in the semen sample.

Preferably, the reaction dyeing device includes a reagent mixer, and the reagent mixer is provided with at least one reagent box for placing at least one reaction reagent and/or staining reagent.

Preferably, the liquid flow system includes a second sheath liquid peristaltic pump, a second buffer filter device, a reaction detection chamber, a waste liquid pool, and a plurality of liquid level sensors; the reaction detection chamber includes a sheath liquid reaction chamber that is fixedly connected in sequence And a laser detection chamber; the dyeing reaction pool is connected to the sheath liquid reaction chamber through a solenoid valve and a transmission tube, and the sheath liquid pool is sequentially connected to the sheath liquid reaction chamber through a second sheath liquid peristaltic pump and a second buffer filter device, and the laser detection chamber It is connected to the waste liquid pool through the transmission tube; multiple liquid level sensors are set in the semen sample pool, sheath liquid pool, cleaning liquid pool, dyeing reaction pool and waste liquid pool for measuring the liquid level;

The dyed sperm sample of the dyeing reaction pool enters the sheath fluid reaction chamber through the transmission tube, and the sheath liquid of the sheath fluid pool enters the sheath fluid reaction chamber through the second sheath fluid peristaltic pump and the second buffer filter device. The dyed sperm sample and sheath fluid are in The sheath fluid sperm sample is formed by the reaction in the sheath fluid reaction chamber. The inner stained sperm sample of the sheath fluid sperm sample presents a linear arrangement of single cells through the laser detection chamber, and is transferred to the waste liquid pool through the transfer tube for recovery processing; the sheath fluid sperm sample is external The layered sheath fluid and the fluid-focused fluid flow of the inner stained sperm sample. Each sperm cell in the stained sperm sample is labeled and stained with a specific fluorescent probe, which can emit fluorescence at a specific wavelength after being excited by the corresponding laser.

Preferably, the laser optical system includes a laser emitting device, a fiber optic tube, a focusing lens, a first beam splitter, a forward light path device, a lateral light path device, and a fluorescence path device;

The laser emitting device, the fiber tube, the focusing lens, the laser detection chamber, and the first dichroic mirror are arranged in an optical path. The focusing lens is arranged on one side of the laser detection chamber, and the forward light path device is arranged on the opposite side of the laser detection chamber. The first dichroic mirror is arranged on the other side of the laser detection room, and the lateral light path device is arranged at an angle to the first dichroic mirror;

The laser light emitted by the laser emitting device is guided to a focusing lens through a fiber tube, and the focusing lens directly irradiates the focused laser light on each sperm cell flowing in the laser detection chamber, and the laser light illuminates each sperm cell to form a forward light signal , The forward optical signal is received by the forward optical path device;

After the focused laser irradiates each sperm cell, a shadow will be formed behind the sperm cell due to its different size, thereby forming a forward light signal, and the forward light signal is received by the forward light path device; After irradiating each sperm cell with laser light, laterally refracted light with different light intensities will be formed on the lateral side of the sperm cell due to the different cell density, thereby forming a lateral light signal, which is refracted by the first beam splitter after being refracted Received to the optical path device, the lateral optical signal is an optical signal lower than Anm;

After the laser beam illuminates the fluorescence of each sperm cell, it will refract different fluorescent signals. After the laser beam illuminates the fluorescent probe on each sperm cell, different labeled probes can be After laser excitation, it will emit fluorescent signals with different characteristic wavelengths;

The fluorescent signal passes through the first beam splitter to form a first fluorescent signal, and the first fluorescent signal is received by the fluorescent channel device; the first fluorescent signal is a fluorescent signal exceeding Anm. Preferably, the fluorescent pathway device includes a second beam splitter, a first fluorescent receiver, a third beam splitter, a second fluorescent receiver, and a third fluorescent receiver;

The first fluorescent signal is irradiated to the second beam splitter to form a second fluorescent signal after passing through, and the second fluorescent signal is received by the first fluorescence receiving device; the first fluorescent signal is irradiated to the second beam splitter through the After being refracted, a third fluorescent signal is formed, the third fluorescent signal is received by a third beam splitter; the second fluorescent signal is a first fluorescent signal lower than Bnm, and the third fluorescent signal is a third fluorescent signal higher than Bnm A fluorescent signal; the third fluorescent signal is irradiated to the third beam splitter to form a fourth fluorescent signal after being refracted, and the fourth fluorescent signal is received by the second fluorescent receiving device; the third fluorescent signal is irradiated to the third After passing through the three-division mirror, a fifth fluorescent signal is formed, and the fifth fluorescent signal is received by a third fluorescent receiving device; the fourth fluorescent signal is a third fluorescent signal lower than Cnm, and the fifth fluorescent signal It is the third fluorescence signal higher than Cnm.

Preferably, the forward light path device is a forward light bandpass filter, and the lateral light path device is a lateral light bandpass filter; the value range of A is 490-510nm, The value range of B is 550-600nm, and the value range of C is 600-640nm; the first fluorescence receiving device is a first fluorescent bandpass filter, and the first fluorescent bandpass filter can pass 500nm -A second fluorescent signal at 550 nm; the second fluorescent receiving device is a second fluorescent band pass filter, and the second fluorescent band pass filter can pass a fourth fluorescent signal at 550 nm-600 nm; the third The fluorescence receiving device is a third fluorescent bandpass filter, and the third fluorescent bandpass filter can pass the fifth fluorescent signal of 600 nm-680 nm.

Preferably, the photoelectric detection system includes a forward light photoelectric converter, a side light photoelectric converter, a first fluorescent photoelectric converter, a second fluorescent photoelectric converter, and a third fluorescent photoelectric converter;

The forward light photoelectric converter is provided behind the forward light path device for converting the forward light signal into a forward electrical signal; the lateral light photoelectric converter is provided behind the lateral light path device, It is used to convert the lateral optical signal into a lateral electrical signal; the first fluorescent photoelectric converter is arranged behind the first fluorescent receiving device, and is used to convert the second fluorescent signal into a second electrical signal; the second The fluorescence photoelectric converter is arranged behind the second fluorescence receiving device and is used to convert the fourth fluorescence signal into a fourth electrical signal; the third fluorescence photoelectric converter is arranged behind the third fluorescence receiving device and is used to convert the first Five fluorescent signals are converted into fifth electrical signals;

Preferably, the control system includes a control device for control, an analog-to-digital converter, a data analysis device, a data storage device, and a data output device all connected to the control device.

The photoelectric detection system converts the received multiple optical signals into electrical signals and sends them to the analog-to-digital converter. The analog-to-digital converter converts the multiple electrical signals into digital signals and stores them in the data storage device, and the data analysis device The digital signal is decoded into corresponding values of various parameters for statistical analysis and analysis results are generated, and the data output device is used to output the analysis results.

Beneficial effect

The implementation of the above technical solution of the automatic sperm cell detector of the present invention has the following advantages or beneficial effects: the semen sample of the automatic sperm cell detector of the present invention enters into the liquid after sample suction, reagent addition, and mixed staining reaction in the sample processing system The flow system drives the semen sample after the dyeing reaction to the detection site under the drive of the flow system. The laser light emitted by the laser optical system is used for excitation detection, and the photoelectric detection system collects the excited optical signal, which is converted into a digital signal by the control system. After data analysis, the sperm quality parameters such as total sperm, density, nuclear integrity, mitochondrial membrane potential, acrosome reaction and other sperm in the semen sample to be detected are calculated according to the data characteristics and cluster analysis algorithm. Fully automated high-throughput rapid analysis of semen samples is realized. As long as the user places the semen samples in the sample buffer pool, they can perform fully automatic operations and output the detection and analysis results, which can be directly connected to the hospital Lis/His system. Medical institutions and third-party inspection institutions at all levels can also connect to WeChat service numbers, website and other back-end databases to achieve seamless connection of data management, easy operation, one-key intelligence, low cost and wide adaptability.

BRIEF DESCRIPTION

In order to more clearly explain the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings used in the description of the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without paying any creative labor, other drawings can be obtained based on these drawings. In the drawings:

FIG. 1 is a schematic diagram of the appearance structure of an embodiment of the automatic sperm cell detector of the present invention;

2 is a schematic diagram of a first decomposition structure of an embodiment of the automatic sperm cell detector of the present invention;

3 is a schematic diagram of a first internal structure of an embodiment of the automatic sperm cell detector of the present invention;

4 is a schematic diagram of a second internal structure of an embodiment of the automatic sperm cell detector of the present invention;

5 is a schematic diagram of a third internal structure of an embodiment of the automatic sperm cell detector of the present invention;

6 is a schematic diagram of a sample counting device of an embodiment of an automatic sperm cell detector of the present invention;

7 is a schematic diagram of a sample processing system of an embodiment of an automatic sperm cell detector of the present invention;

8 is a schematic diagram of a laser optical system of an embodiment of the automatic sperm cell detector of the present invention.

Embodiments of the invention

In order to make the purpose, technical solutions and advantages of the present invention clearer, various exemplary embodiments to be described below will refer to corresponding drawings, which constitute a part of the exemplary embodiments, which describe the implementation of the present invention Various exemplary embodiments that may be employed, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present disclosure. It should be understood that they are only examples of devices and methods consistent with some aspects disclosed in the present invention as detailed in the appended claims, and other embodiments may be used or performed on the embodiments listed herein Structural and functional modifications without departing from the scope and essence of the invention. In other cases, detailed descriptions of well-known systems, devices, circuits, and methods are omitted so as not to obscure the description of the present invention with unnecessary details. In order to explain the technical solution of the present invention, the following will be described with specific embodiments.

1-8 shows a schematic structural diagram provided by an embodiment of the present invention. For ease of description, only parts related to the embodiment of the present invention are shown. An embodiment of a fully automatic sperm cell detector is provided, which includes a test bench 10, a control system 60 disposed on the test bench 10, a sample processing system 20, a fluid flow system 30, and a laser optical system that are all connected to the control system 60 40 and a photoelectric detection system 50; wherein, the sample processing system 20 is used for counting statistics of sperm cells in the semen sample to be processed, and performing a dyeing reaction process on the sperm cells in the semen sample to obtain a dyed sperm sample; The flow system 30 is connected to the sample processing system 20 for mixing, flowing, detecting, and recovering the dyed sperm sample and the sheath fluid output by the sample processing system 20. Specifically, the mixed fluid is the sheath fluid and at least one reaction reagent And/or staining reagents; the control system 60 is used to control the sample processing system 20, the flow system 30, the laser optical system 40, and the photoelectric detection system 50, so as to automatically detect, analyze, and analyze the semen samples. Generate analysis results.

In the automatic sperm cell detector of the present invention, the semen sample enters the liquid flow system after sample preparation, aspiration, reagent addition, mixed staining reaction in the sample processing system, and the semen sample is transferred to the reaction detection chamber under the drive of the liquid flow system After the laser excitation detection by the laser optical system, and the photoelectric detection system collects the excited optical signal, the control system converts it into a digital signal for statistical analysis of the data, and calculates the to-be-detected based on the data characteristics and the unique cluster analysis algorithm Analysis results of total sperm count, density, nuclear integrity, mitochondrial membrane potential, acrosome reaction in the semen samples. The sperm quality analysis method is the core basis for fully automatic analysis and processing of the data collected by the test and issuing results. Fully automated high-throughput rapid analysis of semen samples is realized. As long as the user places the semen samples in the sample pool, they can perform fully automatic operations and output the detection and analysis results, which can be directly connected with the hospital Lis/His system, suitable for all Level medical institutions and third-party inspection institutions can also connect to WeChat service numbers, website and other back-end databases to achieve seamless connection of data management without tedious and complicated operations. After selecting the corresponding detection index, one-key automatic From the processing to the issuance of the results, all are automatically completed. From the sample aspiration processing, the reagent mixing reaction and the fluorescent labeling and other tedious steps, all realize the standard automation. Achieve full-automatic, multi-functional, high-throughput, simple, fast and low-cost clinical detection and analysis of sperm quality analysis.

In this embodiment, the sample processing system 20 includes a semen sample pool 201, a sample counting device 202, a sample injection needle 203, a sheath liquid pool 204, a cleaning liquid pool 205, a reaction dyeing device 206, a plurality of transfer tubes 207, A plurality of solenoid valves 301, a first sheath fluid peristaltic pump 302, a first buffer filter device 303, a sample injection pump 304, a staining reaction cell 305, and an automatic temperature control device 306; specifically, the semen sample pool 201 passes through the A sample counting device 202, a sample injection needle 203, a solenoid valve 301, and a sample injection pump 304 are connected to the staining reaction tank 305. The semen sample pool 201 is used to store a semen sample to be tested. The semen sample includes a large number of sperm cells, specifically The number is not specifically limited here.

Specifically, the sheath liquid pool 204 is used to store the sheath liquid to be mixed, and the cleaning liquid pool 205 is used to store the cleaning liquid. Specifically, the cleaning liquid may be a special cleaning liquid or clean water used to clean the entire detector; The sheath liquid pool 204 and the cleaning liquid pool 205 are both connected to the reaction dyeing device 206 through a transmission tube 207, and the sheath liquid in the sheath liquid pool 204 and the cleaning liquid in the cleaning liquid pool 205 are used for the reaction dyeing after the completion of the dyeing reaction The device 206 and the flow system 30 perform cleaning. The reaction dyeing device 206 is connected to the dyeing reaction tank 305 through a solenoid valve 301 and a first sheath liquid peristaltic pump 302, and the first buffer filter device 303 is used to pulse filter and remove impurities from the sheath liquid output by the sheath liquid pool 204; The reaction dyeing device 206 is used to provide at least one reaction reagent and/or dyeing reagent.

Specifically, the automatic temperature control device 306 is disposed below the dyeing reaction tank 305, and is used to heat the dyeing reaction tank 305 according to the reaction temperature requirements; the reaction reagent and/or the dyeing reagent in the reaction dyeing device 206 are peristaltic pump by the first sheath liquid 302 is sucked out and enters the dyeing reaction tank 305; the semen sample in the semen sample pool 201 enters the dyeing reaction tank 305 through the sample injection pump 304, and the semen sample and at least one reaction reagent and/or dyeing reagent undergo dyeing reaction in the dyeing reaction pool 305 to form Stain sperm samples. Specifically, the automatic temperature control device 306 automatically controls the reaction of the dyeing reaction tank 305 and the temperature and dyeing time during dyeing. After completion, the syringe pump pushes the fully reacted reaction fluid (sperm sample) into the flow system and enters the reaction detection room.

As shown in FIG. 6, the sample counting device 202 is connected to the semen sample pool 201 and the sample injection needle 203, and is used to count and measure the size of each sperm cell in the semen sample passing therethrough; specifically, The sample counting device 202 includes a female electrode 221, a male electrode 222, and a voltage pulse measuring device 223. The voltage pulse measuring device 223 is electrically connected to the female electrode 221 and the male electrode 222; specifically, the female electrode 221 and the male electrode 222 together form a micropore channel 224 for sperm cells to pass through. The micropore channel 224 is set on one side of the semen sample pool 201 and on the other side of the sample injection needle 203; when different sperm cells in the semen sample pass through the microchannel The pore channel 224 generates different voltage pulse signals, and the voltage pulse measuring device 223 measures different voltage pulse signals. The voltage pulse signal includes information on the number and size of sperm cells in the semen sample.

In the present invention, a sample counting device 202 (that is, a flow resistance counting device that uses impedance technology for counting) is provided at the sample injection needle 203 in the sample processing system. The low frequency of the semen sample (sperm cells) filled with phosphate buffer is Electric fields below 5 MHz are non-conductive. The micropore channel 224 of the sample counting device 202 is 20-50um. According to the Coulter cell counting principle, the size and number of each sperm cell in the semen sample drawn by the sample injection needle 203 are measured. When the sperm cells pass through the micropore channel 224, the impedance will increase at this time. This change causes a voltage pulse signal to be generated at the micropore channel. The voltage pulse measurement device 223 will measure different voltage pulse signals. The number of times and pulse changes to record the number and size of sperm cells drawn.

As shown in FIG. 7, the reaction dyeing device 206 includes a reagent mixer 261. The reagent mixer 261 is provided with at least one reagent box 262 for placing at least one reaction reagent and/or staining reagent. The sheath liquid pool 204 and the cleaning liquid pool 205 are both connected to the reaction dyeing device 206 of the reaction dyeing device 206 through a transmission tube 207. The sheath liquid in the sheath liquid pool 204 and the cleaning liquid in the cleaning liquid pool 205 are used to wash and complete the dyeing The reagent mixer 261 and the dyeing reaction tank 305 after the reaction are washed. The mixing device for loading and automatic mixing of reaction reagents and dyeing reagents may be a device for fixing a single test item, or a matrix or circular multi-reagent mixing device for multiple test items. The dyeing reaction cell 305 may be a single device, or a matrix or circular multi-reaction cell integrated device matched with multiple detection items. Specifically, the staining reaction cell 305 can also be a collection of round or square high-throughput multi-staining reaction cells, and connect the semen samples to the mixed solution, which can be used for continuous or simultaneous detection of a large number of high-throughput samples, and the automatic temperature The control device 306 specifically controls the temperature. More specifically, the transmission pipes 207 of the sheath liquid pool 204 and the cleaning liquid pool 205 are each provided with a solenoid valve 301 for controlling the conduction of the switches of the sheath liquid pool 204 and the cleaning liquid pool 205 respectively. More specifically, the reagent mixer 261, the sheath liquid pool 204, and the cleaning liquid pool 205 are all connected to the dyeing reaction tank 305 through a transfer tube 207.

In this embodiment, the liquid flow system 30 includes a second sheath liquid peristaltic pump 309, a second buffer filter device 310, a reaction detection chamber 307, a waste liquid pool 308, and a plurality of liquid level sensors (not shown); Wherein, the reaction detection chamber 307 includes a sheath liquid reaction chamber 371 and a laser detection chamber 372 that are fixedly connected in sequence; the dyeing reaction tank 305 is connected to the sheath liquid reaction chamber 371 through a solenoid valve 301 and a transmission tube 207, and the sheath liquid pool 204 The sheath liquid reaction chamber 371 is connected to the sheath liquid reaction chamber 371 through the second sheath liquid peristaltic pump 309 and the second buffer filter device 310 in sequence, and the laser detection chamber 372 is connected to the waste liquid tank 308 through the transmission pipe 207.

Specifically, the semen sample pool 201 is connected to the dyeing reaction tank 305 through a solenoid valve 301 and a sample injection pump 304; the reaction dyeing device 206 is connected to the dyeing reaction pool through the solenoid valve 301, the first sheath fluid peristaltic pump 302, and the first buffer filter device 303 Connected to 305, the first buffer filtering device 303 is used to perform pulse filtering and impurity filtering on the sheath liquid transferred from the sheath liquid pool 204 through the first sheath liquid peristaltic pump 302.

Specifically, the dyeing reaction cell 305 is connected to the sheath liquid reaction chamber 371 through a solenoid valve 301 and a transfer tube 207, and the sheath liquid pool 204 is sequentially connected to the sheath liquid reaction chamber 371 through a second sheath liquid peristaltic pump 309 and a second buffer filter device 310 The laser detection chamber 372 is connected to the waste liquid pool 308 through the transmission pipe 207, and the waste liquid pool 308 recovers the detected reaction liquid. The specific embodiment of the present invention, but should not be limited to the scheme of the device design.

The present invention also solves the requirement for the integration and automation of the reaction dyeing of semen samples. In the sample processing system, the semen samples are counted and enter the dyeing reaction tank. The specific operation process is as follows: First, the cleaning liquid of the cleaning liquid pool 205 rinses the entire system (transfer tube , Dyeing reaction tank, reaction detection room, etc.), the automatic temperature control device 306 controls the temperature of the dyeing reaction pool to the required temperature, and the semen sample enters the dyeing reaction tank according to the designed amount. At the same time, the reagent mixer automatically controls the reaction reagent and/or dyeing reagent After entering the dyeing reaction tank, after the mixing and reaction time set by the procedure, the dyeing reaction pool controls the dyed sperm sample to be introduced into the sheath liquid reaction chamber 371 of the reaction detection chamber 307 from the transfer tube.

In this embodiment, a plurality of liquid level sensors (not shown) are provided in the semen sample pool 201, the sheath liquid pool 204, the cleaning liquid pool 205, the dyeing reaction pool 305, and the waste liquid pool 308 for measuring their liquids And send corresponding reminder information in real time; the reaction reagent and/or staining reagent in the reaction dyeing device 206 is sucked out by the first sheath fluid peristaltic pump 302 and enters the dyeing reaction tank 305 through the first buffer filter device 303; semen sample The semen sample in the pool 201 enters the staining reaction tank 305 through the sample injection pump 304, and the semen sample and at least one reaction reagent and/or staining reagent are mixed in the staining reaction tank 305 to form a stained sperm sample.

In this embodiment, the dyed sperm sample of the dyeing reaction tank 305 enters the sheath fluid reaction chamber 371 through the transfer tube 207, and the sheath fluid of the sheath fluid pool 204 enters through the second sheath fluid peristaltic pump 309 and the second buffer filter device 310 Sheath fluid reaction chamber 371, according to the principle of liquid focusing, dyed sperm samples and sheath fluid react in the sheath fluid reaction chamber 371 to form a sheath fluid sperm sample, the sheath fluid sperm sample outer sheath fluid sheath fluid sperm sample enveloping the inner sperm cells , And the dyed sperm sample is in the middle of the flow, the inner dyed sperm sample of the sheath fluid sperm sample presents a linear arrangement of single cells through the laser detection chamber 372, and is transmitted to the waste liquid pool 308 through the transfer tube 207 for recovery processing; the sheath fluid sperm sample For the outer sheath fluid and the inner layer of sperm sample fluid focused fluid flow, each sperm cell in the stained sperm sample is labeled and stained with a specific fluorescent probe, and the fluorescent probe can emit fluorescence of a specific wavelength after being excited by the corresponding laser .

Specifically, the sheath fluid is sucked out of the sheath fluid pool by the second sheath fluid peristaltic pump and enters the sheath fluid reaction chamber 371 through the second buffer filter device, and is mixed with the stained sperm sample output from the staining reaction tank to form an external and internal laminar flow. Outside, the sample fluid is inside, and the rapid flow of the outer sheath fluid creates an inward pressure on the slow sperm cells of the inner layer to form a focused linearity of the inner sample detection flow, wherein the stained sperm cells to be detected are arranged in a single linear line through the laser The testing room 372 receives laser testing.

As shown in FIG. 8, the laser optical system 40 includes a laser emitting device 401, a fiber optic tube 402, a focusing lens 403, a first dichroic mirror 404, a forward light path device 405, a lateral light path device 406, and a fluorescence path device 407; The laser emitting device 401, the fiber tube 402, the focusing lens 403, the laser detection chamber 372, and the first dichroic mirror 404 are arranged in the optical path, the focusing lens 403 is arranged on the laser detection chamber 372 side, and the forward light path device 405 is arranged in the laser detection Opposite the chamber 372, the first dichroic mirror 404 is disposed on the other side of the laser detection chamber 372, and the lateral light path device 406 is disposed at an angle to the first dichroic mirror 404.

In this embodiment, the laser light emitted by the laser emitting device 401 is guided to the focusing lens 403 through the optical fiber tube 402, and the focusing lens 403 directly irradiates the focused laser light on each sperm cell flowing in the laser detection chamber 372. The sperm cells will form a shadow behind the sperm cells due to their different sizes, thereby forming a forward light signal, which is received by the forward light path device 405; after the laser irradiates each sperm cell, it will be in the sperm cell. Due to the different density of sperm cells in the lateral direction, laterally refracted light with different intensities is formed to form a lateral optical signal. The lateral optical signal is refracted by the first beam splitter 404 and received by the lateral optical path device 406. The lateral optical signal It is an optical signal lower than Anm.

At the same time, after the laser irradiates the fluorescence of each sperm cell, it will refract different fluorescent signals. After the laser irradiates the fluorescent probe on each sperm cell, different fluorescent probes can be excited by lasers with different characteristic wavelengths. Fluorescent signals with different characteristic wavelengths will be emitted; that is, after the laser is irradiated to the fluorescent probes labeled on each sperm cell, the fluorescent probes will be excited by the laser to emit fluorescent signals with corresponding characteristic wavelengths; specifically, the laser After irradiating each of the fluorescent probes of the sperm cells, the fluorescent probes are excited to emit fluorescent signals with different characteristic wavelengths; the fluorescent signals form the first fluorescent signal after passing through the first dichroic mirror 404, the first A fluorescent signal is received by the fluorescent pathway device 407; the first fluorescent signal is a fluorescent signal whose (wavelength) exceeds Anm.

In this embodiment, the laser emitting device 401 is specifically a semiconductor blue 488 nanometer fiber laser emitter, and the laser emitting device 401 is a semiconductor solid-state laser emitter. The present invention uses a semiconductor solid-state laser emitter as a light source in the laser light path system, and The laser optical fiber is used as the optical path transmission medium, and the optical fiber tube and the focusing lens form the optical path. After the laser is emitted, it is guided to the side of the laser detection chamber through the focusing lens after being guided by the optical fiber, passes through the side wall of the laser detection chamber, and directly illuminates each sperm cell passing through the laser detection chamber, and the forward path of the laser light path passes through the laser detection. The chamber is provided with a forward light path device 406. Among them, the laser optical system involved in the present invention can be a fiber-optic closed conduction optical path, which can make the loading of the laser, filter, and optical signal collection path more flexible and compact, and avoid external light interference, which greatly reduces the loss of light in the optical path compared with the traditional optical path. .

In this embodiment, the fluorescence path device 407 includes a second dichroic mirror 471, a first fluorescence receiving device 472, a third dichroic mirror 473, a second fluorescence receiving device 474, and a third fluorescence receiving device 475; The first fluorescent signal is irradiated to the second dichroic mirror 471 and passes through it to form a second fluorescent signal, and the second fluorescent signal is received by the first fluorescence receiving device 472; A third fluorescent signal is formed, and the third fluorescent signal is received by the third dichroic mirror 473; the second fluorescent signal is the first fluorescent signal with a (wavelength) lower than Bnm, and the third fluorescent signal is with a high (wavelength) The first fluorescent signal at Bnm; the third fluorescent signal is irradiated to the third dichroic mirror 473 to form a fourth fluorescent signal after being refracted, the fourth fluorescent signal is received by the second fluorescent receiving device 474; the third fluorescent signal is illuminated After the third dichroic mirror 473 passes through, a fifth fluorescent signal is formed, and the fifth fluorescent signal is received by the third fluorescent receiving device 475; the fourth fluorescent signal is a third fluorescent signal (wavelength) lower than Cnm, so The fifth fluorescence signal is a third fluorescence signal (wavelength) higher than Cnm. Specifically, the value range of A (wavelength) is 490-510 nm, the value range of B (wavelength) is 550-600 nm, and the value range of C (wavelength) is 600-640 nm.

In this embodiment, forward scattered light: the characteristic light signal formed by the size of the sperm cells in front of the optical path after the laser irradiates the sperm cells; side scattered light: the side of the optical path due to the sperm after the laser irradiates the sperm cells Characteristic light signals formed by different cell densities; Fluorescence: After the laser irradiates the sperm cells, the fluorescent probes labeled on the sperm cells are excited, thereby emitting fluorescent signals with different characteristic wavelengths. Streaming optical channel: each channel of optical signal after flow laser detection can receive this kind of optical signal. Different instruments and equipment are set differently. The detector of the present invention is equipped with FS channel (forward scattered light channel for receiving forward direction) Optical signal), SS channel (side scatter light channel, used to receive side light signal), FITC channel (FITC fluorescent channel, used to receive FITC fluorescent signal), JC1 channel (JC1 fluorescent channel, used to receive JC1 fluorescent signal ), AO channel (AO fluorescence channel, used to receive AO fluorescence signal) / PI channel (PI fluorescence channel, used to receive PI fluorescence signal).

In this embodiment, the forward light path device 405 is a forward light bandpass filter, which is an FS channel bandpass filter, and the lateral light path device 406 is a lateral light bandpass filter, which is SS channel bandpass filter; the first fluorescent receiver 472 is the first fluorescent bandpass filter, which is the FITC fluorescent channel bandpass filter, the first fluorescent bandpass filter can pass (wavelength) 500nm- The second fluorescent signal at 550nm, that is, the FITC fluorescent signal; the second fluorescent receiving device 474 is a second fluorescent bandpass filter, that is, a JC1 fluorescent channel bandpass filter, and the second fluorescent bandpass filter can pass ( Wavelength) 550nm-600nm fourth fluorescence signal, namely JC1 fluorescence signal; the third fluorescence receiving device 475 is the third fluorescence bandpass filter, that is, AO fluorescence channel bandpass filter or PI fluorescence channel bandpass filter The third fluorescent bandpass filter can pass the (wavelength) 600nm-680nm fifth fluorescent signal, which is the AO fluorescent signal or PI fluorescent signal.

In this embodiment, the photoelectric detection system 50 includes a forward light photoelectric converter 501, a side light photoelectric converter 502, a first fluorescent photoelectric converter 503, a second fluorescent photoelectric converter 504, and a third fluorescent photoelectric conversion 505; the forward light photoelectric converter 501, which is provided behind the forward light path device 405, is used to convert the forward light signal into a forward electrical signal; the side light photoelectric converter 511, is provided on the side Behind the light path device 406, it is used to convert the lateral light signal into a lateral electrical signal; the first fluorescent photoelectric converter 512, which is provided behind the first fluorescent receiving device 472, is used to convert the second fluorescent signal into A second electrical signal; the second fluorescence photoelectric converter 513, which is arranged behind the second fluorescence receiving device 474, and is used to convert the fourth fluorescence signal into a fourth electrical signal; and the third fluorescence photoelectric converter 514, which is arranged After the third fluorescent receiving device 475, it is used to convert the fifth fluorescent signal into a fifth electrical signal; the control system 60 includes a control device 601 for control, an analog-to-digital converter 602 connected to the control device 601, The data analysis device 603, the data storage device 604, and the data output device 605.

The photoelectric detection system 50 converts the received multiple optical signals (forward electrical signal, lateral electrical signal, second fluorescence signal (FITC fluorescence signal), fourth fluorescence signal (JC1 fluorescence signal), and fifth fluorescence signal ( AO fluorescent signal or PI fluorescent signal)) are converted into electrical signals (forward electrical signals, lateral electrical signals, second electrical signals (FITC electrical signals), fourth electrical signals (JC1 electrical signals), and fifth electrical signals ( AO electrical signal or PI electrical signal)) and sent to the analog-to-digital converter 602 of the control system 60, the analog-to-digital converter 602 converts multiple electrical signals into digital signals and stores them in the data storage device 604, the data analysis device 603 pairs Multiple digital signals are decoded into corresponding values of various parameters for statistical analysis and analysis results are generated, and the data output device 605 is used to output the analysis results. Specifically, the photoelectric converter receives the optical signal and reacts into an electrical signal according to the strength, and then converts it into digital by the analog-to-digital converter and stores it in the data storage device 604. Specifically, the photoelectric signals detected by the detector are all converted into digital signals and stored in the range of 2 ^0-2 ¹⁰ (1-1024). Among them, the control system 60 adopts the single-chip MSP430 of Texas Instruments (TI) as the core control of the system architecture, the analog-to-digital converter 602 adopts 6-channel high-speed and high-precision synchronous acquisition of optical signals based on the ADS5560 analog-to-digital conversion chip, and the data storage device adopts 27C512 storage The chip is then analyzed and calculated by the MSP430 main control system. Of course, other companies and other series of chips or components with the same or similar functions can also be used. The specifics are not limited here.

In this embodiment, the control system 60 analyzes and identifies the data, specifically including: standard data reading, data judgment and identification method, and standard format data storage; it involves a standard format conversion of the data results obtained by the test, and instrument and sample test information The standard data format and code, and the method of format code identification and conversion, the storage format and extraction method of the standard format data of all information.

In this embodiment, the multi-parameter cluster analysis includes: sperm shape feature cluster model analysis, sperm cell nuclear structure feature cluster model analysis, sperm mitochondrial membrane potential feature cluster model analysis, sperm acrosomal membrane structure feature cluster model analysis 、Sperm acrosome reaction characteristic cluster model analysis, anti-sperm antibody characteristic cluster model analysis, semen white blood cell shape characteristic cluster model analysis, semen white blood cell peroxidase characteristic cluster model analysis, sperm cell peroxidase characteristic cluster Group model analysis; specifically, based on the data characteristics of different quality parameters, a mathematical analysis model of corresponding parameters is established. When clustering analysis of sample parameters, the corresponding variable data is taken according to different parameters. First, each variable data is standardized according to the characteristics of the data matrix Correct, and then use the corresponding mathematical model for cluster reference analysis.

In this embodiment, single cell feature analysis includes: effective cell data reading according to different parameter models, single cell parameter analysis with different quality parameter models, and single cell different quality parameter data record storage; specifically, clustering parameters according to different parameters Compare the results of the analysis, determine the effective cell data to identify each parameter, calculate the variable data for each effective cell, and record the corresponding results. Sperm population characteristic statistics include: determination and statistics of all single-cell characteristics of different quality parameters, statistical analysis of cell population characteristics of different quality parameters, and data storage of different quality parameter data of cell groups. Specifically, it is the item-by-item determination and statistics of the calculation results of all single-cell characteristics under each quality parameter, and the calculation and description of the statistical characteristics of all effective cell populations of the corresponding quality parameters, and the corresponding results are recorded. The calculation results and analysis report include: single cell parameter result calculation in semen samples, cell population parameter result calculation in semen samples, semen sample quality analysis data recording and storage, semen sample quality analysis report, specifically all semen samples obtained The recording and storage of each parameter result and the analysis report template are completed and a report is issued.

In this embodiment, the control system is mainly completed by the circuit integration system in the instrument. The storage, analysis and calculation procedures contained in the chip of the circuit automatically complete the automatic processing, storage, analysis and calculation of the data after detection, and according to the template Save or output the report file in spreadsheet format that conforms to the Lis/His system, or the result file in other database, graphics, and picture formats for transmission to a variety of media needs, such as website background, WeChat background, etc., and can also be sent directly to network printers Perform a direct print result report.

The automatic sperm cell detector of the present invention is a new medical-grade clinical detection instrument and equipment, which relates to flow cytometry technology, micro-automated cytochemical reaction processing technology, clustering mathematical model intelligent comparison automatic calculation technology, using the Coulter cell counting principle, Flow focusing technology and fluorescent labeling technology quickly obtain the parameters of each sperm cell in the sperm cell population, and calculate the total number of sperm cells, density, nuclear integrity, mitochondrial membrane potential, acrosomal membrane integrity, Induces acrosome reaction, sperm viable cell rate, sperm cell apoptosis rate, semen white blood cell count, leukocyte reactive oxygen species, sperm reactive oxygen species and other parameters that reflect the quality of sperm cells. Each test can automatically analyze up to tens of thousands of sperm cells in the sample one by one, which significantly improves the accuracy and accuracy of the test results and is very stable. It is used in the field of sperm quality detection and analysis and is very suitable for human medical institutions at all levels The clinical testing needs can also be used for animal sperm quality testing and analysis in the animal husbandry industry.

The above are only preferred embodiments of the present invention, and those skilled in the art will easily think of other embodiments of the present disclosure after considering the description and practicing the invention disclosed herein. Those skilled in the art are aware that various changes or equivalent replacements can be made to these features and embodiments without departing from the spirit and scope of the present invention. In addition, under the teaching of the present invention, any variations, uses, or adaptive changes can be made to these features and embodiments. These variations, uses, or adaptive changes follow the general principles of the present disclosure and include the technology not disclosed in the present disclosure. Common knowledge or common technical means in the field will not depart from the spirit and scope of the present invention. Therefore, the description and the embodiments are only regarded as exemplary, and the present invention is not limited by the specific embodiments disclosed herein, and all the embodiments falling within the scope of the claims of the present application belong to the protection scope of the present invention.

Claims

A fully automatic sperm cell detector, characterized in that it includes a detection stand (10), a control system (60) provided on the detection stand (10), and both are connected to the control system (60) Sample processing system (20), liquid flow system (30), laser optical system (40) and photoelectric detection system (50);

The sample processing system (20) is used for counting statistics of sperm cells in the semen sample to be processed, and performing a dyeing reaction process on the sperm cells in the semen sample to obtain a dyed sperm sample; the fluid flow system (30) Connected to the sample processing system (20), for mixing, flowing, detecting and recovering the dyed sperm sample and sheath fluid output from the sample processing system (20);

The control system (60) is used to control the sample processing system (20), the liquid flow system (30), the laser optical system (40) and the photoelectric detection system (50), so as to fully automate the semen sample Detect, analyze and generate analysis results.
The automatic sperm cell detector according to claim 1, wherein the sample processing system (20) includes a semen sample pool (201), a sample counting device (202), a sample injection needle (203), a sheath Liquid pool (204), cleaning liquid pool (205), reaction dyeing device (206), multiple transfer tubes (207), multiple solenoid valves (301), first sheath liquid peristaltic pump (302), first buffer filtration A device (303), a sample injection pump (304), a staining reaction cell (305) and an automatic temperature control device (306); the semen sample cell (201) passes through the sample counting device (202) and the sample injection needle in sequence (203), a solenoid valve (301) and a sample injection pump (304) are connected to the staining reaction tank (305), the semen sample pool (201) is used to store the semen sample to be tested, the semen sample Including a large number of sperm cells;

The sample counting device (202) is connected to the semen sample pool (201) and the sample injection needle (203), and is used to count each sperm cell in the semen sample passing therethrough and Size measurement

The sheath liquid pool (204) is used to store the sheath liquid to be mixed, and the cleaning liquid pool (205) is used to store cleaning liquid; the sheath liquid pool (204) and the cleaning liquid pool (205) are both passed The transfer tube (207) is connected to the reaction dyeing device (206), and the sheath liquid in the sheath liquid pool (204) and the cleaning liquid in the cleaning liquid pool (205) are used to complete the dyeing reaction The reaction dyeing device (206) and the liquid flow system (30) for cleaning; the reaction dyeing device (206) passes through the solenoid valve (301), the first sheath liquid peristaltic pump (302) and the A dyeing reaction tank (305) is connected, and the first buffer filtering device (303) is used for pulse filtering and impurity filtering of the sheath liquid output by the sheath liquid pool (204); the reaction dyeing device (206), For providing at least one reaction reagent and/or staining reagent;

The automatic temperature control device (306) is arranged under the dyeing reaction tank (305), and is used for heating the dyeing reaction tank (305) according to the reaction temperature requirements; The reaction reagent and/or staining reagent are sucked out by the first sheath fluid peristaltic pump (302) and enter the staining reaction pool (305); the semen sample in the semen sample pool (202) passes through the sample A syringe pump (304) enters the dyeing reaction pool (305), and the semen sample and at least one of the reaction reagent and/or dyeing reagent undergo a dyeing reaction in the dyeing reaction pool (305) to form a dyed sperm sample.
The automatic sperm cell detector according to claim 2, characterized in that the sample counting device (202) includes a negative electrode (221), a positive electrode (222), a voltage pulse measuring device (223), the voltage The pulse measuring device (223) is electrically connected to the negative electrode (221) and the positive electrode (222); the negative electrode (221) and the positive electrode (222) together form a passage for the sperm cells to pass through Micropore channel (224), the micropore channel (224) is provided on one side of the semen sample pool (201) and on the other side of the sample injection needle (203); when the semen sample Different sperm cells will generate different voltage pulse signals when they pass through the micropore channel (224), and the voltage pulse measurement device (223) will measure different voltage pulse signals, the voltage pulse signals It includes information on the number and size of sperm cells in the semen sample.
The automatic sperm cell detector according to claim 2, characterized in that the reaction staining device (206) includes a reagent mixer (261), and at least one of the reagent mixer (261) is provided for placing A kit (262) of at least one reaction reagent and/or staining reagent.
The automatic sperm cell detector according to claim 4, characterized in that the liquid flow system (30) includes a second sheath fluid peristaltic pump (309), a second buffer filtration device (310), and a reaction detection chamber ( 307), a waste liquid pool (308) and a plurality of liquid level sensors; the reaction detection chamber (307) includes a sheath liquid reaction chamber (371) and a laser detection chamber (372) that are fixedly connected in sequence;

The dyeing reaction tank (305) is connected to the sheath liquid reaction chamber (371) through the solenoid valve (301) and the transfer tube (207), and the sheath liquid pool (204) sequentially passes through the second sheath liquid A peristaltic pump (309) and a second buffer filter device (310) are connected to the sheath liquid reaction chamber (371), and the laser detection chamber (372) is connected to the waste liquid pool (308) through the transfer tube (207) ) Connection; a plurality of the liquid level sensors are respectively arranged in the semen sample pool (201), sheath liquid pool (204), cleaning liquid pool (205), dyeing reaction pool (305) and waste liquid pool (308) Used to measure its liquid level; the dyed sperm sample of the dyeing reaction cell (305) enters the sheath fluid reaction chamber (371) through the transfer tube (207), and the sheath of the sheath fluid pool (204) The fluid enters the sheath fluid reaction chamber (371) through the second sheath fluid peristaltic pump (309) and the second buffer filter device (310). The dyed sperm sample and the sheath fluid are in the sheath fluid reaction chamber (371) The internal reaction forms a sheath fluid sperm sample. The inner layer stained sperm sample of the sheath fluid sperm sample presents a linear arrangement of single cells through the laser detection chamber (372) and is transmitted to the laboratory via the transfer tube (207) The waste liquid pool (308) is recycled; the sheath fluid sperm sample is a fluid-focused fluid flow of the outer sheath fluid and the inner stained sperm sample, and each sperm cell in the stained sperm sample is specifically fluorescent Probe dyeing treatment, after being excited by the corresponding laser, can emit fluorescence with a specific wavelength.
The automatic sperm cell detector according to claim 5, wherein the laser optical system (40) includes a laser emitting device (401), a fiber tube (402), a focusing lens (403), and a first spectroscopic reflector Mirror (404), forward light path device (405), side light path device (406), fluorescence path device (407); the laser emitting device (401), fiber optic tube (402), focusing lens (403) 1. A laser detection chamber (372), a first beam splitter (404) are arranged in an optical path, the focusing lens (403) is arranged on the side of the laser detection chamber (372), and the forward light path device (405) Disposed on the opposite side of the laser detection chamber (372), the first dichroic mirror (404) is disposed on the other side of the laser detection chamber (372), the lateral light path device (406) and the The first dichroic mirror (404) is arranged at an angle; the laser light emitted by the laser emitting device (401) is guided to the focusing lens (403) through the fiber tube (402), and the focusing lens (403) will focus The irradiated laser directly irradiates each of the sperm cells flowing in the laser detection chamber (372); after the focused laser irradiates each of the sperm cells, the sperm cells will be behind because of their different sizes A shadow is formed, thereby forming a forward light signal, which is received by the forward light path device (405); after the laser irradiates each of the sperm cells, it will be caused by the lateral direction of the sperm cells. Different cell densities form laterally refracted light with different light intensities, thereby forming a lateral light signal, which is refracted by the first beam splitter (404) and then refracted by the lateral light path device (406) Receive, the lateral optical signal is an optical signal lower than Anm;

After the focused laser irradiates the fluorescence of each sperm cell, different fluorescent signals will be refracted. After the laser irradiates each fluorescent probe on the sperm cell, different fluorescent probes may be different After being excited by the laser with the characteristic wavelength, it will emit fluorescent signals with different characteristic wavelengths; the fluorescent signal forms a first fluorescent signal after passing through the first spectroscopic mirror (404), and the first fluorescent signal is emitted by the fluorescent The channel device (407) receives; the first fluorescent signal is the fluorescent signal exceeding Anm.
The automatic sperm cell detector according to claim 6, characterized in that the fluorescent pathway device (407) includes a second spectroscopic mirror (471), a first fluorescence receiving device (472), and a third spectroscopic mirror (473), a second fluorescent receiving device (474) and a third fluorescent receiving device (475); the first fluorescent signal is irradiated to the second dichroic mirror (471) to form a second fluorescent signal after passing through, The second fluorescent signal is received by the first fluorescent receiving device (472); the first fluorescent signal is irradiated to the second dichroic mirror (471) to form a third fluorescent signal after being refracted, the first Three fluorescent signals are received by the third beam splitter (473); the second fluorescent signal is the first fluorescent signal lower than Bnm, and the third fluorescent signal is the first fluorescence higher than Bnm A signal; the third fluorescent signal is irradiated to the third dichroic mirror (473) and is refracted to form a fourth fluorescent signal, and the fourth fluorescent signal is received by the second fluorescent receiving device (474); The third fluorescent signal is irradiated to the third dichroic mirror (473) and passes therethrough to form a fifth fluorescent signal, the fifth fluorescent signal is received by the third fluorescent receiving device (475); the fourth The fluorescence signal is the third fluorescence signal lower than Cnm, and the fifth fluorescence signal is the third fluorescence signal higher than Cnm.
The automatic sperm cell detector according to claim 7, wherein the forward light path device (405) is a forward light bandpass filter, and the lateral light path device (406) is a side Bandpass filter; the value range of A is 490-510nm, the value range of B is 550-600nm, the value range of C is 600-640nm; the first fluorescence receiving device (472 ) Is a first fluorescent bandpass filter, the first fluorescent bandpass filter can pass the second fluorescent signal of 500nm-550nm;

The second fluorescent receiving device (474) is a second fluorescent bandpass filter, and the second fluorescent bandpass filter can pass the fourth fluorescent signal of 550nm-600nm; the third fluorescent receiving device (475) is a third fluorescent bandpass filter, and the third fluorescent bandpass filter can pass the fifth fluorescent signal of 600 nm-680 nm.
The automatic sperm cell detector according to claim 7, wherein the photoelectric detection system (50) includes a forward light photoelectric converter (501), a side light photoelectric converter (502), and a first fluorescence A photoelectric converter (503), a second fluorescent photoelectric converter (504) and a third fluorescent photoelectric converter (505);

The forward light photoelectric converter (501) is disposed behind the forward light path device (405) and is used to convert the forward light signal into a forward electrical signal; the side light photoelectric converter (511), which is arranged behind the lateral light path device (406), and is used to convert the lateral optical signal into a lateral electrical signal;

The first fluorescent photoelectric converter (512) is disposed behind the first fluorescent receiving device (472) and is used to convert the second fluorescent signal into a second electrical signal; the second fluorescent photoelectric converter (513), which is arranged behind the second fluorescence receiving device (474), and is used to convert the fourth fluorescence signal into a fourth electrical signal; the third fluorescence photoelectric converter (514) is arranged on the Behind the third fluorescence receiving device (475), it is used to convert the fifth fluorescence signal into a fifth electrical signal.
The automatic sperm cell detector according to claim 9, characterized in that the control system (60) includes a control device (601) for control, and an analog-to-digital converter connected to the control device (601) Device (602), data analysis device (603), data storage device (604) and data output device (605); the photoelectric detection system (50) converts the received multiple optical signals into electrical signals and sends to The analog-to-digital converter (602), the analog-to-digital converter (602) converts a plurality of the electrical signals into digital signals and stores them in the data storage device (604), the data analysis device (603) ) Decode the digital signal into corresponding values of various parameters for statistical analysis and generate analysis results, and the data output device (605) is used to output the analysis results.

A