CN109612911B - Full-automatic sperm cell detector - Google Patents
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- CN109612911B CN109612911B CN201811599058.7A CN201811599058A CN109612911B CN 109612911 B CN109612911 B CN 109612911B CN 201811599058 A CN201811599058 A CN 201811599058A CN 109612911 B CN109612911 B CN 109612911B
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Classifications
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- G—PHYSICS
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- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a full-automatic sperm cell detector, which is characterized in that a semen sample enters a liquid flow system after being subjected to sample suction, reagent addition and mixed reaction dyeing, the semen sample after reaction dyeing is transmitted to a reaction detection chamber by the liquid flow system, laser excitation detection is carried out by a laser optical system, an excited light signal is collected by a photoelectric detection system, a control system is used for carrying out data statistics analysis after converting the light signal into a digital signal, and sperm quality parameter results such as total number, density, nuclear integrity, mitochondrial membrane potential, acrosome reaction and the like in the semen sample to be detected are calculated according to data characteristics and a clustering analysis algorithm. The full-automatic high-throughput rapid detection and analysis of the semen sample are realized, a user can perform full-automatic operation only by placing the semen sample in the sample tank, and automatically output detection and analysis results, and the full-automatic high-throughput rapid detection and analysis device can be directly in butt joint with a hospital LiS/His system to realize seamless connection of data management, is simple and convenient to operate, is intelligent by one key, is low in cost and has wide adaptability.
Description
Technical Field
The invention relates to the technical field of novel medical-grade clinical detection, in particular to a full-automatic sperm cell detector.
Background
Sperm quality assessment is a key to diagnosing fertility in males (males), and detection items include sperm individual or population indicators such as sperm density, motility, deformity, nuclear integrity, mitochondrial membrane potential, acrosome response, and the like. At present, main detection equipment comprises a microscope, a fluorescence microscope, a general flow cytometer and the like, and main detection methods comprise manual microscopic examination of the microscope, computer-aided sperm analysis system (CASA) aided analysis and computer-aided manual analysis of a flow cytometer standard cell analysis system.
The microscope manual microscopic examination is the most mainstream application technology at present, and mainly adopts a microscope to carry out manual naked eye counting, vitality distinguishing and morphological observation after the semen cell smear is prepared, and also has a computer-aided system to assist in analyzing images obtained by the microscope. The main defects are that a large amount of manpower is required, if a computer-aided analysis system is added, the detection cost is greatly increased, and most importantly, only a small amount of indexes such as the number, the activity and the morphology of sperms can be detected, other important indexes such as the integrity of sperm nuclei, the mitochondrial function, the acrosome membrane and the like are required to be marked by a fluorescent probe after cell pictures are prepared, then the fluorescent probe is adopted for microscopic analysis, and at present, no automatic analysis auxiliary software passing clinical authentication is required, so that the cost is high, and a large amount of manpower is required to be consumed for manual visual observation, discrimination and counting.
The method for detecting and analyzing the universal flow cytometer has been applied to the detection of clinical indexes of sperms, and the flow cytometer has great technical advantages in the aspect of rapidly detecting a large number of cells, can realize the accurate detection of high-flux samples, greatly expands the index parameters of the clinical sperms with detectable quality, and is widely focused by clinical workers. However, the flow cytometer is mainly imported equipment, is expensive, is not provided with a special instrument for detecting spermatids in the world at present, needs long-time special training in the actual use process, needs various indexes such as manual voltage adjustment, compensation, flow rate and the like in the detection process, is provided with various related areas or doors, is difficult to meet the objectivity, the repeatability and the referential performance of clinical detection requirements, also needs manual area and door screening division in the detection completion process of analysis results, and is difficult to meet the daytime repeatability and the room referential performance of the clinical detection requirements.
Most of flow cytometry is a universal detection platform or a multi-laser multi-channel multi-parameter instrument for scientific research, in a specific application scene, sample processing is usually separated from detection, and an instrument for automatically completing the integration of automatic sample processing, reaction dyeing and sample injection detection is not available internationally, so that the universal clinical flow cytometry is more suitable for scientific research or a small amount of blood cell index detection with higher detection requirement.
At present, the existing instrument is lack of a full-automatic integrated high-throughput low-cost detection instrument special for sperm cell special analysis in clinical semen detection at home and abroad, and sample treatment, instrument detection and result analysis brought by the existing instrument are all carried out in terms and all need manual operation, so that the detection operation is complex, the manual sample treatment step is complex, and the equipment maintenance and operation problems are required to be carried out by special training.
Disclosure of Invention
The invention aims to solve the technical problem of providing a full-automatic sperm cell detector aiming at the defects in the prior art.
The technical scheme adopted for solving the technical problems is as follows: according to an aspect of the present invention, there is provided a full-automatic sperm cell tester, including a testing bench, a control system disposed on the testing bench, a sample processing system, a liquid flow system, a laser optical system, and a photoelectric detection system all connected to the control system; specifically, the sample processing system is used for counting and counting the sperm cells in the semen sample to be processed and performing dyeing reaction on the sperm cells in the semen sample to obtain a dyed sperm sample; the liquid flow system is connected with the sample processing system and is used for carrying out mixed reaction, flow, detection and recovery treatment on the stained sperm sample output by the sample processing system and sheath liquid; the control system is used for controlling the sample processing system, the liquid flow system, the laser optical system and the photoelectric detection system, so as to perform full-automatic detection and analysis on the semen sample and generate an analysis result;
The photoelectric detection system comprises a forward photoelectric converter, a lateral photoelectric converter, a first fluorescent photoelectric converter, a second fluorescent photoelectric converter and a third fluorescent photoelectric converter;
the forward optical-to-electrical converter is arranged behind the forward optical access device and is used for converting a forward optical signal into a forward electrical signal;
the lateral photoelectric converter is arranged behind the lateral optical path device and is used for converting a lateral optical signal into a lateral electric signal;
the first fluorescent photoelectric converter is arranged behind the first fluorescent receiving device and is used for converting a second fluorescent signal into a second electric signal;
the second fluorescent photoelectric converter is arranged behind the second fluorescent receiving device and is used for converting a fourth fluorescent signal into a fourth electric signal;
the third fluorescent photoelectric converter is arranged behind the third fluorescent receiving device and is used for converting a fifth fluorescent signal into a fifth electric signal.
Preferably, the sample processing system comprises a semen sample pool, a sample counting device, a sample injection needle, a sheath liquid pool, a cleaning liquid pool, a reaction dyeing device, a plurality of transmission pipes, a plurality of electromagnetic valves, a first sheath liquid peristaltic pump, a first buffer filtering device, a sample injection pump, a dyeing reaction pool and an automatic temperature control device;
The semen sample pool is connected with the dyeing reaction pool sequentially through a sample counting device, a sample injection needle, an electromagnetic valve and a sample injection pump, and is used for storing a semen sample to be detected, wherein the semen sample comprises a large number of spermatids;
the sample counting device is connected with the semen sample pool and the sample injection needle and is used for counting and measuring the size of each sperm cell in the semen sample passing through the sample counting device;
the sheath liquid pool is used for storing sheath liquid to be mixed, and the cleaning liquid pool is used for storing cleaning liquid; the sheath liquid in the sheath liquid 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 dyeing reaction is completed;
the reaction dyeing device is connected with the dyeing reaction tank through an electromagnetic valve and a first sheath liquid peristaltic pump, and the first buffer filtering device is used for carrying out pulse filtration and impurity filtration on sheath liquid output by the sheath liquid tank; a reaction staining device for providing at least one reaction reagent and/or staining reagent;
the automatic temperature control device is arranged below the dyeing reaction tank and is used for heating the dyeing reaction tank according to the reaction temperature requirement;
The reaction reagent and/or the dyeing reagent in the reaction dyeing device are pumped out by a peristaltic pump of the primary sheath liquid and enter a dyeing reaction tank; the semen sample in the semen sample pool enters a staining reaction pool through a sample injection pump, and the semen sample and at least one reaction reagent and/or staining reagent carry out staining reaction in the staining reaction pool to form a stained sperm sample.
Preferably, the sample counting device comprises a negative electrode, a positive electrode and a voltage pulse measuring device, wherein the voltage pulse measuring device is electrically connected with the negative electrode and the positive electrode;
the negative electrode and the positive electrode jointly form a micropore channel for passing sperm cells, one side of the micropore channel is arranged in the semen sample pool, and the other side of the micropore channel is arranged in the sample injection needle;
when different sperm cells in the semen sample pass through the micropore channel, different voltage pulse signals are generated, and the voltage pulse measuring device can measure the different voltage pulse signals, wherein the voltage pulse signals comprise the number and the size information of the sperm cells in the semen sample.
Preferably, the reaction staining apparatus comprises a reagent mixer provided with at least one reagent kit for placing at least one reaction reagent and/or staining reagent.
Preferably, the liquid flow system comprises a second sheath peristaltic pump, a second buffer filtering device, a reaction detection chamber, a waste liquid tank and a plurality of liquid level sensors; the reaction detection chamber comprises a sheath liquid reaction chamber and a laser detection chamber which are fixedly communicated in sequence;
the dyeing reaction tank is connected with the sheath liquid reaction chamber through an electromagnetic valve and a transmission pipe, the sheath liquid tank is connected with the sheath liquid reaction chamber through a second sheath liquid peristaltic pump and a second buffer filter device in sequence, and the laser detection chamber is connected with the waste liquid tank through the transmission pipe;
the liquid level sensors are respectively arranged in the semen sample pool, the sheath liquid pool, the cleaning liquid pool, the dyeing reaction pool and the waste liquid pool and used for measuring the liquid level of the liquid level sensors;
the method comprises the steps that a dyeing sperm sample in a dyeing reaction tank enters a sheath liquid reaction chamber through a transmission pipe, sheath liquid in a sheath liquid pool enters the sheath liquid reaction chamber through a second sheath liquid peristaltic pump and a second buffer filtering device, the dyeing sperm sample and the sheath liquid react in the sheath liquid reaction chamber to form a sheath liquid sperm sample, and an inner layer dyeing sperm sample of the sheath liquid sperm sample shows single cell linear arrangement, passes through a laser detection chamber and is transmitted to a waste liquid pool through the transmission pipe for recovery treatment; the sheath liquid sperm sample is fluid focusing liquid flow of outer sheath liquid and inner staining sperm sample, each sperm cell in the staining sperm sample is marked and stained by a specific fluorescent probe, and fluorescence with specific wavelength can be emitted after the sperm cell is excited by corresponding laser.
Preferably, the laser optical system comprises a laser emitting device, an optical fiber tube, a focusing lens, a first beam splitting reflector, a forward light path device, a lateral light path device and a fluorescence path device;
the laser emission device, the optical fiber tube, the focusing lens, the laser detection chamber and the first light splitting reflector are arranged in a light path, the focusing lens is arranged on one side of the laser detection chamber, the forward light path device is arranged on the opposite side of the laser detection chamber, the first light splitting reflector is arranged on the other side of the laser detection chamber, and the lateral light path device and the first light splitting reflector are arranged at an angle;
the laser emitted by the laser emitting device is guided to the focusing lens through the optical fiber tube, the focusing lens directly irradiates focused laser onto each sperm cell flowing in the laser detection chamber, a forward optical signal is formed after the laser irradiates each sperm cell, and the forward optical signal is received by the forward optical access device;
the laser after focusing irradiates each sperm cell to form shadows behind the sperm cell due to the different sizes of the sperm cells, thereby forming a forward optical signal, and the forward optical signal is received by the forward optical access device; after each sperm cell is irradiated by the laser, lateral refraction light with different light intensities is formed on the sperm cell in the lateral direction due to different cell densities, so that a lateral light signal is formed, the lateral light signal is received by a lateral light path device after being refracted by a first light splitting reflector, and the lateral light signal is an optical signal lower than Anm;
After the laser irradiates the fluorescent probes on each sperm cell, the different labeled probes can be excited by the laser with different characteristic wavelengths, and then fluorescent signals with different characteristic wavelengths can be emitted;
the fluorescent signal passes through a first light-splitting reflector to form a first fluorescent signal, and the first fluorescent signal is received by a fluorescent access device; the first fluorescent signal is a fluorescent signal exceeding Anm.
Preferably, the fluorescence path device comprises a second light splitting reflector, a first fluorescence receiving device, a third light splitting reflector, a second fluorescence receiving device and a third fluorescence receiving device;
the first fluorescent signal irradiates a second light-splitting reflector to pass through the second light-splitting reflector to form a second fluorescent signal, and the second fluorescent signal is received by a first fluorescent receiving device; the first fluorescent signal irradiates the second light-splitting reflector and is refracted by the second light-splitting reflector to form a third fluorescent signal, and the third fluorescent signal is received by the third light-splitting reflector; the second fluorescent signal is a first fluorescent signal lower than Bnm, and the third fluorescent signal is a first fluorescent signal higher than Bnm;
The third fluorescent signal irradiates the third light-splitting reflector and is refracted by the third light-splitting reflector to form a fourth fluorescent signal, and the fourth fluorescent signal is received by the second fluorescent receiving device; the third fluorescent signal irradiates the third light-splitting reflector to pass through the third light-splitting reflector to form a fifth fluorescent signal, and the fifth fluorescent signal is received by the third fluorescent receiving device; the fourth fluorescent signal is a third fluorescent signal below Cnm and the fifth fluorescent signal is a third fluorescent signal above Cnm.
Preferably, the forward optical path device is a forward optical band-pass filter, and the lateral optical path device is a lateral optical band-pass filter;
the numerical range of A is 490-510nm, the numerical range of B is 550-600nm, and the numerical range of C is 600-640nm;
the first fluorescence receiving device is a first fluorescence band-pass filter, and the first fluorescence band-pass filter can pass through second fluorescence signals of 500nm-550 nm;
the second fluorescence receiving device is a second fluorescence band-pass filter, and the second fluorescence band-pass filter can pass a fourth fluorescence signal of 550nm-600 nm;
the third fluorescence receiving device is a third fluorescence band-pass filter, and the third fluorescence band-pass filter can pass a fifth fluorescence signal of 600nm-680 nm.
Preferably, the control system comprises a control device for control, an analog-to-digital converter, a data analysis device, a data storage device and a data output device which are all connected with the control device.
The photoelectric detection system respectively converts the received optical signals into electric signals and sends the electric signals to the analog-to-digital converter, the analog-to-digital converter converts the electric signals into digital signals and stores the digital signals in the data storage device, the data analysis device decodes the digital signals into corresponding numerical values of all parameters for statistical analysis and generates analysis results, and the data output device is used for outputting the analysis results.
The implementation of the technical scheme of the full-automatic sperm cell detector has the following advantages or beneficial effects: the semen sample of the full-automatic sperm cell detector enters a liquid flow system after sample suction, reagent addition and mixed dyeing reaction in a sample processing system, the semen sample after dyeing reaction is transmitted to a detection part under the driving of the liquid flow system, the laser excitation detection is carried out through the laser optical system, the light signal after excitation is collected by a photoelectric detection system, the data analysis is carried out after the light signal is converted into a digital signal by a control system, and the sperm quality parameter results such as the total number, the density, the nuclear integrity, the mitochondrial membrane potential, the acrosome reaction and the like in the semen sample to be detected are calculated according to the data characteristics and a clustering analysis algorithm. The full-automatic high-throughput rapid analysis of semen samples is realized, a user can perform full-automatic operation only by placing semen samples in a sample buffer pool, and output detection analysis results, and the full-automatic high-throughput rapid analysis device can be directly connected with a hospital LiS/His system, is suitable for various medical institutions and third-party inspection institutions, can also be connected with background databases such as micro-communication service numbers, websites and the like, realizes seamless connection of data management, is simple and convenient to operate, is intelligent by one key, has low cost and wide adaptability.
Drawings
For a clearer description of the technical solutions of embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art, in which:
FIG. 1 is a schematic view of the external appearance of an embodiment of a fully automated sperm cell tester of the present invention;
FIG. 2 is a schematic diagram of a first exploded view of an embodiment of a fully automated sperm cell tester of the present invention;
FIG. 3 is a schematic diagram of a first internal structure of an embodiment of a fully automated sperm cell tester of the present invention;
FIG. 4 is a schematic diagram of a second internal structure of an embodiment of a fully automated sperm cell tester of the present invention;
FIG. 5 is a schematic diagram of a third internal structure of an embodiment of a fully automated sperm cell tester of the present invention;
FIG. 6 is a schematic diagram of a sample counting device of an embodiment of a fully automated sperm cell tester of the present invention;
FIG. 7 is a schematic diagram of a sample processing system of an embodiment of a fully automated sperm cell tester of the present invention;
FIG. 8 is a schematic diagram of a laser optical system of an embodiment of the fully automated sperm cell tester of the present invention.
Detailed Description
For a better understanding of the objects, technical solutions and advantages of the present utility model, reference should be made to the various exemplary embodiments described hereinafter with reference to the accompanying drawings, which form a part hereof, and in which are shown by way of illustration various exemplary embodiments in which the utility model may be practiced, and in which like numerals in the various figures designate identical or similar elements unless otherwise indicated. The implementations described in the following exemplary examples are not representative of all implementations consistent with the present disclosure. It is to be understood that they are merely examples of apparatus and methods consistent with certain aspects of the present disclosure as set forth in the appended claims, other embodiments may be utilized, or structural and functional modifications may be made to the embodiments set forth herein, without departing from the scope and spirit of the present disclosure. In other instances, detailed descriptions of well-known systems, devices, circuits, and methods are omitted so as not to obscure the description of the present utility model with unnecessary detail.
In order to illustrate the technical scheme of the utility model, the following description is made by specific examples.
Fig. 1 to 8 show schematic structural diagrams provided by the embodiment of the present utility model, and only parts relevant to the embodiment of the present utility model are shown for convenience of explanation. A fully automatic sperm cell tester embodiment is provided, comprising a test bench 10, a control system 60 disposed on the test bench 10, a sample processing system 20, a liquid flow system 30, a laser optical system 40, and a photoelectric detection system 50, all connected to the control system 60; the sample processing system 20 is used for counting the sperm cells in the semen sample to be processed and performing dyeing reaction on the sperm cells in the semen sample to obtain a dyed sperm sample; the liquid flow system 30 is connected with the sample processing system 20, and is used for performing mixed reaction, flow, detection and recovery processing on the stained sperm sample output by the sample processing system 20 and sheath liquid, and specifically, the mixed liquid is formed by mixing the sheath liquid with at least one reaction reagent and/or staining reagent; the control system 60 is used for controlling the sample processing system 20, the liquid flow system 30, the laser optical system 40 and the photoelectric detection system 50, and further performing full-automatic detection and analysis on the semen sample and generating an analysis result.
In the full-automatic sperm cell detector, a semen sample enters a liquid flow system after sample preparation, absorption, reagent addition and mixed dyeing reaction in a sample processing system, the semen sample is transmitted to a reaction detection chamber under the driving of the liquid flow system, the excited light signals are collected by a photoelectric detection system and are converted into digital signals by a control system, data statistics analysis is carried out, and analysis results such as the total number, density, nuclear integrity, mitochondrial membrane potential, acrosome reaction and the like of the semen sample to be detected are calculated according to data characteristics and a unique aggregation analysis algorithm. The sperm quality analysis method is a core basis for carrying out full-automatic analysis processing on the data collected by detection and outputting results. The full-automatic high-throughput rapid analysis of the semen sample is realized, a user can perform full-automatic operation only by placing the semen sample in a sample pool, and output detection analysis results, the full-automatic operation can be directly connected with a hospital LiS/His system, the full-automatic detection device is suitable for various medical institutions and third-party inspection institutions, background databases such as micro-signal service numbers, websites and the like can also be connected, seamless connection of data management is realized, the corresponding detection indexes are not required to be selected, one-key full-automatic operation is performed, all automatic completion is performed from sample processing to result, and all complicated steps such as sample sucking processing, reagent mixing reaction and fluorescent marking are realized, so that standard automatic formation is realized. The full-automatic, multifunctional, high-flux, simple, convenient, quick and low-cost clinical detection and analysis of sperm quality analysis are realized.
In this embodiment, the sample processing system 20 includes a semen sample cell 201, a sample counting device 202, a sample injection needle 203, a sheath fluid cell 204, a cleaning fluid cell 205, a reaction staining device 206, a plurality of transfer tubes 207, a plurality of solenoid valves 301, a first sheath fluid peristaltic pump 302, a first buffer filtering 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 is connected to the staining reaction pool 305 sequentially through the sample counting device 202, the sample injection needle 203, the electromagnetic valve 301 and the sample injection pump 304, where the semen sample pool 201 is used for storing a semen sample to be detected, and the semen sample includes a large number of sperm cells, and the specific number is not limited herein.
Specifically, the sheath liquid pool 204 is used for storing sheath liquid to be mixed, the cleaning liquid pool 205 is used for storing cleaning liquid, and specifically, the cleaning liquid can be special cleaning liquid or clean water used for cleaning the whole detector; the sheath fluid tank 204 and the cleaning fluid tank 205 are connected to the reaction dyeing device 206 through a transmission pipe 207, and the sheath fluid in the sheath fluid tank 204 and the cleaning fluid in the cleaning fluid tank 205 are used for cleaning the reaction dyeing device 206 and the liquid flow system 30 after the dyeing reaction is completed. The reaction dyeing device 206 is connected with a dyeing reaction tank 305 through an electromagnetic valve 301 and a first sheath liquid peristaltic pump 302, and the first buffer filtering device 303 is used for performing pulse filtration and impurity filtration on the sheath liquid output by the sheath liquid tank 204; the reaction staining apparatus 206 is configured to provide at least one reaction reagent and/or staining reagent.
Specifically, the automatic temperature control device 306 is disposed below the dyeing reaction tank 305, and is configured to heat the dyeing reaction tank 305 according to a reaction temperature requirement; the reactive and/or staining reagents in the reactive staining apparatus 206 are aspirated by the primary sheath peristaltic pump 302 and enter the staining reaction tank 305; the semen sample in the semen sample tank 201 enters a staining reaction tank 305 through a sample injection pump 304, and the semen sample and at least one reaction reagent and/or staining reagent undergo a staining reaction in the staining reaction tank 305 to form a stained sperm sample. Specifically, the automatic temperature control device 306 automatically controls the reaction of the dyeing reaction tank 305, the temperature during dyeing and the reaction dyeing time, and after the completion, the reaction liquid (semen sample) with complete reaction dyeing is pushed into the liquid flow system by the injection pump and enters the reaction detection chamber.
As shown in fig. 6, the sample counting device 202 is connected to the semen sample cell 201 and the sample injection needle 203, and is used for counting and measuring the size of each sperm cell in the semen sample passing through the sample counting device; specifically, the sample counting device 202 includes a negative electrode 221, a positive electrode 222, and a voltage pulse measuring device 223, where the voltage pulse measuring device 223 is electrically connected to both the negative electrode 221 and the positive electrode 222; specifically, the negative electrode 221 and the positive electrode 222 together form a microporous channel 224 for passing sperm cells, one side of the microporous channel 224 is disposed in the semen sample pool 201, and the other side is disposed in the sample injection needle 203; when different sperm cells in the semen sample pass through the microporous channel 224, different voltage pulse signals are generated, and the voltage pulse measuring device 223 measures the different voltage pulse signals, wherein the voltage pulse signals comprise the number and the size information of the sperm cells in the semen sample.
The present invention provides a sample counting device 202 (i.e., a flow impedance counting device, which counts using impedance techniques) at the sample injection needle 203 in a sample processing system, and the semen sample (sperm cells) is rendered non-conductive in an electric field at a low frequency of 5MHz or less, which is filled with phosphate buffer. The micro-porous channel 224 of the sample counting device 202 is 20-50um, and the size and number of each sperm cell in the semen sample sucked by the sample injection needle 203 are measured according to the coulter cell counting principle. When sperm cells pass through the microporous passageway 224, the impedance will rise at this time, and this change causes a voltage pulse signal to be generated at the microporous passageway, and the voltage pulse measuring device 223 will measure different voltage pulse signals, and record the number and size information of the sucked sperm cells according to the occurrence times and pulse changes of the voltage pulse signals.
In this embodiment, as shown in fig. 7, the reaction staining apparatus 206 includes a reagent mixer 261, and at least one reagent kit 262 for placing at least one reaction reagent and/or staining reagent is disposed on the reagent mixer 261. The sheath fluid tank 204 and the cleaning fluid tank 205 are connected to the reaction dyeing device 206 of the reaction dyeing device 206 through the transmission pipe 207, and the sheath fluid in the sheath fluid tank 204 and the cleaning fluid in the cleaning fluid tank 205 are used for cleaning the reagent mixer 261 and the dyeing reaction tank 305 after the dyeing reaction is completed. The mixing device for loading and automatically mixing the reaction reagent and the dyeing reagent can be a device for fixing a detection item singly or a matrix or circular mixing device for loading multiple reagents matched with multiple detection items. The dyeing reaction tank 305 can be a single device, or can be used for continuously or simultaneously detecting a large number of samples matched with a plurality of detection items, and the temperature is specifically controlled by the automatic temperature control device 306. More specifically, the delivery pipes 207 of the sheath fluid reservoir 204 and the cleaning fluid reservoir 205 are respectively provided with an electromagnetic valve 301 for controlling the on-off of the sheath fluid reservoir 204 and the cleaning fluid reservoir 205, respectively. More specifically, the reagent mixer 261, the sheath fluid reservoir 204, and the wash fluid reservoir 205 are all connected to the staining reaction tank 305 through a transfer line 207.
In this embodiment, the liquid flow system 30 includes a second sheath peristaltic pump 309, a second buffer filter device 310, a reaction detection chamber 307, a waste liquid reservoir 308, and a plurality of liquid level sensors (not shown); wherein, the reaction detection chamber 307 comprises a sheath liquid reaction chamber 371 and a laser detection chamber 372 which are fixedly communicated in sequence; the dyeing reaction tank 305 is connected with the sheath liquid reaction chamber 371 through the electromagnetic valve 301 and the transmission pipe 207, the sheath liquid tank 204 is connected with 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 with the waste liquid tank 308 through the transmission pipe 207.
Specifically, the semen sample cell 201 is connected with a staining reaction cell 305 through an electromagnetic valve 301 and a sample injection pump 304; the reaction dyeing device 206 is connected with the dyeing reaction tank 305 through an electromagnetic valve 301, a first sheath peristaltic pump 302 and a first buffer filtering device 303, and the first buffer filtering device 303 is used for performing pulse filtration and impurity filtration on the sheath liquid transferred from the sheath liquid tank 204 through the first sheath peristaltic pump 302.
Specifically, the dyeing reaction tank 305 is connected with the sheath liquid reaction chamber 371 through the electromagnetic valve 301 and the transmission pipe 207, the sheath liquid tank 204 is connected with the sheath liquid reaction chamber 371 sequentially through the second sheath liquid peristaltic pump 309 and the second buffer filter device 310, the laser detection chamber 372 is connected with the waste liquid tank 308 through the transmission pipe 207, and the waste liquid tank 308 recovers the detected reaction liquid. The specific embodiments of the invention are not limited to this device design.
The invention simultaneously solves the requirement of integrated automation of semen sample reaction and dyeing, and in a sample processing system, semen samples enter a dyeing reaction tank after being counted, and the specific operation flow is as follows: firstly, the cleaning solution in the cleaning solution tank 205 washes the whole system (transmission pipe, dyeing reaction tank, reaction detection chamber, etc.), the automatic temperature control device 306 controls the temperature of the dyeing reaction tank to the required temperature, the semen sample enters the dyeing reaction tank according to the design amount, meanwhile, the reagent mixer automatically controls the reaction reagent and/or the dyeing reagent to enter the dyeing reaction tank, and after the mixing and reaction time made by the program, the dyeing reaction tank controls the dyeing sperm sample to be guided into the sheath solution reaction chamber 371 of the reaction detection chamber 307 by the transmission pipe.
In this embodiment, a plurality of liquid level sensors (not shown) are respectively disposed in the semen sample pool 201, the sheath pool 204, the cleaning liquid pool 205, the dyeing reaction pool 305 and the waste liquid pool 308 for measuring the liquid levels thereof and sending corresponding reminding information in real time; the reaction reagent and/or the dyeing reagent in the reaction dyeing device 206 is sucked out by the primary sheath peristaltic pump 302 and enters the dyeing reaction tank 305 through the primary buffer filtering device 303; the semen sample in the semen sample cell 201 enters the staining reaction cell 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 cell 305 to form a stained sperm sample.
In this embodiment, the stained sperm sample in the stain reaction tank 305 enters the sheath liquid reaction chamber 371 through the transmission pipe 207, the sheath liquid in the sheath liquid pond 204 enters the sheath liquid reaction chamber 371 through the second sheath liquid peristaltic pump 309 and the second buffer filter 310, according to the liquid focusing principle, the stained sperm sample and the sheath liquid react in the sheath liquid reaction chamber 371 to form a sheath liquid sperm sample, the sheath liquid sperm sample of the inner sperm cell is wrapped by the sheath liquid sperm sample of the outer sheath liquid sperm cell of the sheath liquid sperm cell, and the stained sperm sample is in the middle of the liquid flow, the inner stained sperm sample of the sheath liquid sperm sample presents single cell linear arrangement and passes through the laser detection chamber 372, and is transmitted to the waste liquid pond 308 through the transmission pipe 207 for recovery treatment; the sheath liquid sperm sample is fluid focusing liquid flow of an outer sheath liquid and an inner staining sperm sample, each sperm cell in the staining sperm sample is marked and stained by a specific fluorescent probe, and the fluorescent probe can emit fluorescence with specific wavelength after being excited by corresponding laser.
Specifically, sheath liquid is sucked out of the sheath liquid pond by a second sheath liquid peristaltic pump and enters the sheath liquid reaction chamber 371 through a second buffer filtering device, and is mixed with the stained sperm sample output by the staining reaction pond to form an outer-inner laminar flow, the sheath liquid is outside, the sample liquid is inside, the rapid flow of the outer sheath liquid forms inward pressure on the slow sperm cells of the inner layer to form focusing linearity of inner layer sample detection flow, wherein the stained sperm cells to be detected are arranged in a single line and pass through the laser detection chamber 372, and laser detection is received.
In this embodiment, as shown in fig. 8, the laser optical system 40 includes a laser light emitting device 401, an optical fiber tube 402, a focusing lens 403, a first light dividing mirror 404, a forward light path device 405, a side light path device 406, and a fluorescent light path device 407; the laser emission device 401, the optical fiber tube 402, the focusing lens 403, the laser detection chamber 372 and the first light splitting reflector 404 are arranged in an optical path, the focusing lens 403 is arranged on one side of the laser detection chamber 372, the forward light path device 405 is arranged on the opposite side of the laser detection chamber 372, the first light splitting reflector 404 is arranged on the other side of the laser detection chamber 372, and the lateral light path device 406 is arranged at an angle with the first light splitting reflector 404.
In this embodiment, the laser emitted by the laser emitting device 401 is guided to the focusing lens 403 through the optical fiber tube 402, the focusing lens 403 irradiates the focused laser directly onto each sperm cell flowing in the laser detecting chamber 372, and after irradiating each sperm cell, the laser forms a shadow behind the sperm cell due to the different sizes of the laser, so as to form a forward optical signal, and the forward optical signal is received by the forward optical path device 405; the laser irradiates each sperm cell to form lateral refraction light with different light intensities on the sperm cell lateral direction due to different sperm cell densities, so as to form a lateral light signal, and the lateral light signal is received by the lateral light path device 406 after being refracted by the first light splitting reflector 404, and the lateral light signal is an optical signal lower than Anm.
Meanwhile, after the laser irradiates the fluorescent probes on each sperm cell, the fluorescent probes can be excited by the laser with different characteristic wavelengths, and then the fluorescent signals with different characteristic wavelengths can be emitted; after laser irradiates the fluorescent probes marked on each sperm cell, the fluorescent probes are excited by the laser, so that fluorescent signals with corresponding characteristic wavelengths are emitted; specifically, after the laser irradiates the fluorescent probe of each sperm cell, the fluorescent probe is excited so as to emit fluorescent signals with different characteristic wavelengths; the fluorescence signal passes through the first beam-splitting reflector 404 to form a first fluorescence signal, and the first fluorescence signal is received by the fluorescence channel device 407; the first fluorescent signal is a fluorescent signal (wavelength) exceeding am.
In this embodiment, the laser emitting device 401 is a semiconductor blue light 488 nanometer fiber laser emitter, and the laser emitting device 401 is a semiconductor solid laser emitter, and in the laser light path system, the semiconductor solid laser emitter is used as a light source, and a laser fiber is used as a light path transmission medium, and a fiber tube and a focusing lens form a light path. The laser is guided by the optical fiber after being emitted, irradiates to the side of the laser detection chamber through the focusing lens, passes through the side wall of the laser detection chamber, directly irradiates on each sperm cell passing through the laser detection chamber, and a forward light path device 406 is arranged on the forward path of the laser path passing through the laser detection chamber. The laser optical system can be an optical fiber closed conduction optical path, so that the loading of a laser, a filter and an optical signal collecting path is more flexible and compact, the interference of external light is avoided, and the loss of light in the optical path is greatly reduced compared with the traditional optical path.
In the present embodiment, the fluorescent path means 407 includes a second light-splitting mirror 471, a first fluorescent receiving means 472, a third light-splitting mirror 473, a second fluorescent receiving means 474, and a third fluorescent receiving means 475; the first fluorescent signal irradiates the second beam splitter 471 to pass through the second beam splitter to form a second fluorescent signal, and the second fluorescent signal is received by the first fluorescent receiving device 472; the first fluorescent signal irradiates the second light-splitting reflector 471 and is refracted by the second light-splitting reflector to form a third fluorescent signal, and the third fluorescent signal is received by the third light-splitting reflector 473; the second fluorescent signal is a first fluorescent signal (wavelength) lower than Bnm, and the third fluorescent signal is a first fluorescent signal (wavelength) higher than Bnm; the third fluorescent signal irradiates the third light splitting reflector 473 and is refracted by the third light splitting reflector to form a fourth fluorescent signal, and the fourth fluorescent signal is received by the second fluorescent receiving device 474; the third fluorescent signal irradiates the third light splitting mirror 473 and passes through the third light splitting mirror 473 to form a fifth fluorescent signal, which is received by the third fluorescent receiving device 475; the fourth fluorescent signal is a third fluorescent signal (wavelength) below Cnm and the fifth fluorescent signal is a third fluorescent signal (wavelength) above Cnm. Specifically, the numerical range of A is 490-510nm, the numerical range of B is 550-600nm, and the numerical range of C is 600-640nm.
In this embodiment, the forward scattered light: characteristic optical signals formed by the different sizes of the spermatids right in front of the optical path after the spermatids are irradiated by laser; side scattered light: characteristic optical signals formed by different sperm cell densities at the side of the optical path after the sperm cells are irradiated by laser; fluorescence: after the laser irradiates the sperm cells, the fluorescent probes marked on the sperm cells are excited, so that fluorescent signals with different characteristic wavelengths are emitted. Flow optical channel: the flow laser detection device comprises a flow laser detection device, a flow laser detection device and a flow laser detection device, wherein the flow laser detection device comprises a channel for receiving various optical signals after the flow laser detection, the flow laser detection device can receive the various optical signals, and different instruments and equipment are provided with a FS channel (forward scattering optical channel for receiving forward optical signals), a SS channel (side scattering optical channel for receiving side optical signals), a FITC channel (FITC fluorescent channel for receiving FITC fluorescent signals), a JC1 channel (JC 1 fluorescent channel for receiving JC1 fluorescent signals), an AO channel (AO fluorescent channel for receiving AO fluorescent signals)/PI channel (PI fluorescent channel for receiving PI fluorescent signals).
In this embodiment, the forward optical path device 405 is a forward optical bandpass filter, i.e. an FS channel bandpass filter, and the lateral optical path device 406 is a lateral optical bandpass filter, i.e. an SS channel bandpass filter; the first fluorescence receiving device 472 is a first fluorescence band-pass filter, namely a FITC fluorescence channel band-pass filter, and the first fluorescence band-pass filter can pass a second fluorescence signal with the wavelength of 500nm-550nm, namely a FITC fluorescence signal; the second fluorescent receiving device 474 is a second fluorescent bandpass filter, namely a JC1 fluorescent channel bandpass filter, and the second fluorescent bandpass filter can pass a fourth fluorescent signal with a wavelength of 550nm-600nm, namely a JC1 fluorescent signal; the third fluorescence receiving device 475 is a third fluorescence bandpass filter, namely an AO fluorescence channel bandpass filter or a PI fluorescence channel bandpass filter, and the third fluorescence bandpass filter can pass a fifth fluorescence signal with the wavelength of 600nm-680nm, namely an AO fluorescence signal or a PI fluorescence signal.
In the present embodiment, the photodetection system 50 includes a forward photoelectric converter 501, a side photoelectric converter 502, a first fluorescent photoelectric converter 503, a second fluorescent photoelectric converter 504, and a third fluorescent photoelectric converter 505; the forward optical-to-optical converter 501 is disposed behind the forward optical path device 405 and is configured to convert a forward optical signal into a forward electrical signal; the lateral optical-to-electrical converter 511 is disposed behind the lateral optical access device 406, and is configured 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 configured to convert the second fluorescent signal into a second electrical signal; the second fluorescent photoelectric converter 513 is disposed behind the second fluorescent receiving device 474, and is configured to convert a fourth fluorescent signal into a fourth electrical signal; the third fluorescent photoelectric converter 514 is disposed behind the third fluorescent receiving device 475, and is configured to convert the fifth fluorescent signal into a fifth electrical signal; the control system 60 comprises a control device 601 for controlling, an analog-to-digital converter 602, a data analysis device 603, a data storage device 604 and a data output device 605 which are connected with the control device 601.
The photodetection system 50 converts the received optical signals (forward electric signal, lateral electric signal, second fluorescent signal (FITC fluorescent signal), fourth fluorescent signal (JC 1 fluorescent signal), fifth fluorescent signal (AO fluorescent signal or PI fluorescent signal)) into electric signals (forward electric signal, lateral electric signal, second electric signal (FITC electric signal), fourth electric signal (JC 1 electric signal), fifth electric signal (AO electric signal or PI electric signal)) respectively, and sends the electric signals to the analog-to-digital converter 602 of the control system 60, the analog-to-digital converter 602 converts the electric signals into digital signals and stores the digital signals in the data storage device 604, the data analysis device 603 performs statistical analysis on the decoded digital signals into corresponding values of the respective parameters and generates analysis results, and the data output device 605 is used for outputting the analysis results. Specifically, the photoelectric converter receives the optical signal, reacts to an electrical signal according to intensity, is converted to a digital by the analog-to-digital converter, and is stored by the data storage device 604. Specifically, all photoelectric signals detected by the detector are 2 0 -2 10 The (1-1024) range is converted into a digital signal and stored. The control system 60 uses a single chip microcomputer MSP430 of a Texas Instrument (TI) as a system architecture core control, the analog-to-digital converter 602 uses a 6-channel high-speed high-precision synchronous acquisition optical signal based on an ADS5560 analog-to-digital conversion chip, the data storage device uses a 27C512 storage chip, and the MSP430 main control system performs data analysis and calculation, which can also be used by other companies, other series of chips or components with the same or similar functions, and is not limited in particular herein.
In this embodiment, the control system 60 performs analysis and identification on the data, and specifically includes: standard data reading, data judging and identifying method and standard format data storage; the method is characterized by comprising a standard format conversion step, a standard format and code step, a format code identification conversion step, a standard format data storage form and an extraction method of all information.
In this embodiment, the multiparameter clustering analysis includes: sperm shape characteristic cluster model analysis, sperm cell nuclear structure characteristic cluster model analysis, sperm mitochondrial membrane potential characteristic cluster model analysis, sperm acrosome membrane structure characteristic cluster model analysis, sperm acrosome reaction characteristic cluster model analysis, anti-sperm antibody characteristic cluster model analysis, sperm leukocyte shape characteristic cluster model analysis, sperm leukocyte peroxidase characteristic cluster model analysis, sperm cell internal peroxidase characteristic cluster model analysis; the method comprises the steps of establishing mathematical analysis models of corresponding parameters according to data characteristics of different quality parameters, taking corresponding variable data according to different parameters when sample parameter clustering analysis is carried out, carrying out standardized correction on each variable data according to data matrix characteristics, and carrying out clustering reference analysis by using the corresponding mathematical models.
In this embodiment, the single cell characterization includes: reading effective cell data according to different parameter models, analyzing single cell parameters of different quality parameter models, and recording and storing single cell different quality parameter data; specifically, according to the results of cluster reference analysis of different parameters, the effective cell data of each parameter is judged and identified, the variable data of each effective cell and the corresponding calculation are carried out, and the corresponding results are recorded.
In this embodiment, sperm population characteristic statistics include: and (3) judging and counting all single cell characteristics of different quality parameters, performing cell population characteristic statistical analysis of different quality parameters, and recording and storing data of different quality parameters of the cell population. Specifically, the method comprises the steps of judging and counting item by item for all single cell characteristic calculation results under each quality parameter, calculating and describing statistical characteristics of all effective cell populations of corresponding quality parameters, and recording corresponding results.
In this embodiment, the calculation result and the analysis report include: calculating single cell parameter results in the semen sample, calculating cell population parameter results in the semen sample, recording and storing semen sample quality analysis data, reporting semen sample quality analysis, specifically recording and storing all obtained semen sample parameter results, and completing and reporting an analysis report template.
In this embodiment, the control system is mainly a circuit integrated system in the instrument, and is implemented by a storage, analysis and calculation program contained in a chip in the circuit, automatically completes automatic processing, storage, analysis and calculation of detected data, and stores or outputs an electronic form report file conforming to a list/His system or other result files in a database, a graph and a picture form according to a template for transmission to various media, such as a website background, a WeChat background and the like, and can also directly send a printer available on a network to directly print a result report.
The invention discloses a full-automatic sperm cell detector, which is novel medical-grade clinical detection instrument equipment, and relates to a flow cytometry technology, a micro-automatic cytochemical reaction processing technology and an intelligent comparison automatic calculation technology of a clustered mathematical model. The method can automatically analyze tens of thousands of sperm cells in the sample one by one every time, remarkably improves the accuracy and the accuracy of detection results, is very stable, is applied to the field of sperm quality detection and analysis, is very suitable for clinical detection requirements of all levels of medical institutions of human beings, and can also be used for animal sperm quality detection and analysis in the livestock breeding industry.
The foregoing is merely a preferred example of the present invention and other embodiments of the present disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the spirit and scope of the invention. Further, any modifications, use, or adaptations of the features and embodiments following the general principles of the present disclosure and including such departures from the present disclosure as come within known or customary practice within the art to which the disclosure pertains without departing from the spirit and scope of the invention. Therefore, it is intended that the specification and examples be considered as exemplary only, with the invention not being limited to the particular examples disclosed, but that the invention will include all embodiments falling within the scope of the appended claims.
Claims (9)
1. The full-automatic sperm cell detector is characterized by comprising a detection rack (10), a control system (60) arranged on the detection rack (10), a sample processing system (20), a liquid flow system (30), a laser optical system (40) and a photoelectric detection system (50), wherein the sample processing system (20), the liquid flow system (30), the laser optical system (40) and the photoelectric detection system (50) are connected with the control system (60);
The sample processing system (20) is used for counting and counting the sperm cells in the semen sample to be processed and performing dyeing reaction on the sperm cells in the semen sample to obtain a dyed sperm sample; the liquid flow system (30) is connected with the sample processing system (20) and is used for carrying out mixed reaction, flowing, detection and recovery treatment on the stained sperm sample output by the sample processing system (20) and sheath liquid;
the control system (60) is used for controlling 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 automatically detect, analyze and generate an analysis result on the semen sample;
the photoelectric detection system (50) comprises a forward photoelectric converter (501), a lateral photoelectric converter (502), a first fluorescent photoelectric converter (503), a second fluorescent photoelectric converter (504) and a third fluorescent photoelectric converter (505);
the forward photoelectric converter (501) is arranged behind the forward optical path device (405) and is used for converting a forward optical signal into a forward electric signal;
the lateral photoelectric converter (502) is arranged behind the lateral optical path device (406) and is used for converting a lateral optical signal into a lateral electric signal;
The first fluorescent photoelectric converter (503) is arranged behind the first fluorescent receiving device (472) and is used for converting a second fluorescent signal into a second electric signal;
the second fluorescent photoelectric converter (504) is arranged behind the second fluorescent receiving device (474) and is used for converting a fourth fluorescent signal into a fourth electric signal;
the third fluorescent photoelectric converter (505) is arranged behind the third fluorescent receiving device (475) and is used for converting a fifth fluorescent signal into a fifth electric signal.
2. The fully automated sperm cell tester of claim 1, wherein the sample processing system (20) comprises a semen sample cell (201), a sample counting device (202), a sample injection needle (203), a sheath fluid cell (204), a cleaning fluid cell (205), a reaction staining device (206), a plurality of transfer tubes (207), a plurality of solenoid valves (301), a first sheath fluid peristaltic pump (302), a first buffer filtration device (303), a sample injection pump (304), a staining reaction cell (305), and an automatic temperature control device (306);
the semen sample pool (201) is connected with the dyeing reaction pool (305) sequentially through the sample counting device (202), the sample injection needle (203), the electromagnetic valve (301) and the sample injection pump (304), the semen sample pool (201) is used for storing the semen sample to be detected, and the semen sample comprises a large number of spermatids;
The sample counting device (202) is connected with the semen sample pool (201) and the sample injection needle (203) and is used for counting and measuring the size of each sperm cell in the semen sample passing through the sample counting device;
the sheath liquid pool (204) is used for storing sheath liquid to be mixed, and the cleaning liquid pool (205) is used for storing cleaning liquid; the sheath liquid pool (204) and the cleaning liquid pool (205) are connected to the reaction dyeing device (206) through the transmission pipe (207), and sheath liquid in the sheath liquid pool (204) and cleaning liquid in the cleaning liquid pool (205) are used for cleaning the reaction dyeing device (206) and the liquid flow system (30) after dyeing reaction is completed;
the reaction dyeing device (206) is connected with the dyeing reaction tank (305) through the electromagnetic valve (301) and a first sheath liquid peristaltic pump (302), and the first buffer filtering device (303) is used for performing pulse filtration and impurity filtration on sheath liquid output by the sheath liquid tank (204); -said reaction staining means (206) for providing at least one of a reactive reagent and a staining reagent;
the automatic temperature control device (306) is arranged below the dyeing reaction tank (305) and is used for heating the dyeing reaction tank (305) according to the reaction temperature requirement;
The reaction reagent and the staining reagent in the reaction staining device (206) are sucked out by the first sheath liquid peristaltic pump (302) and enter the staining reaction tank (305); the semen sample in the semen sample pool (202) enters the dyeing reaction pool (305) through the sample injection pump (304), and the semen sample and at least one of the reaction reagent and the dyeing reagent perform dyeing reaction in the dyeing reaction pool (305) to form a dyed semen sample.
3. The fully automatic sperm cell tester as described in claim 2, wherein said sample counting device (202) comprises a negative electrode (221), a positive electrode (222), and a voltage pulse measuring device (223), said voltage pulse measuring device (223) being electrically connected to both said negative electrode (221) and said positive electrode (222);
the negative electrode (221) and the positive electrode (222) jointly form a micropore channel (224) for the spermatid to pass through, one side of the micropore channel (224) is arranged in the semen sample pool (201), and the other side of the micropore channel is arranged in the sample injection needle (203);
when different sperm cells in the semen sample pass through the micropore channel (224), different voltage pulse signals are generated, the voltage pulse measuring device (223) can measure the different voltage pulse signals, and the voltage pulse signals comprise the number and the size information of the sperm cells in the semen sample.
4. The fully automated sperm cell tester as described in claim 2, wherein said reaction staining device (206) comprises a reagent mixer (261), said reagent mixer (261) having at least one reagent cartridge (262) for placing at least one of a reaction reagent and a staining reagent thereon.
5. The fully automated sperm cell tester as described in claim 4, wherein said fluid flow system (30) comprises a second sheath peristaltic pump (309), a second buffer filter device (310), a reaction detection chamber (307), a waste reservoir (308), and a plurality of fluid level sensors;
the reaction detection chamber (307) comprises a sheath liquid reaction chamber (371) and a laser detection chamber (372) which are fixedly communicated in sequence;
the dyeing reaction tank (305) is connected with the sheath liquid reaction chamber (371) through the electromagnetic valve (301) and the transmission pipe (207), the sheath liquid tank (204) is connected with 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 with the waste liquid tank (308) through the transmission pipe (207);
the liquid level sensors are respectively arranged 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) and used for measuring the liquid level of the liquid;
The stained sperm sample in the staining reaction tank (305) enters the sheath liquid reaction chamber (371) through the transmission pipe (207), the sheath liquid in the sheath liquid pond (204) enters the sheath liquid reaction chamber (371) through the second sheath liquid peristaltic pump (309) and the second buffer filtering device (310), the stained sperm sample and the sheath liquid react in the sheath liquid reaction chamber (371) to form a sheath liquid sperm sample, and the inner layer stained sperm sample of the sheath liquid sperm sample is in single-cell linear arrangement, passes through the laser detection chamber (372) and is transmitted to the waste liquid pond (308) through the transmission pipe (207) for recovery treatment; the sheath liquid sperm sample is fluid focusing liquid flow of an outer sheath liquid and an inner staining sperm sample, each sperm cell in the staining sperm sample is marked and stained by a specific fluorescent probe, and fluorescence with specific wavelength can be emitted after the sperm cell is excited by corresponding laser.
6. The fully automated sperm cell tester as described in claim 5, wherein said laser optical system (40) comprises a laser emitting device (401), a fiber tube (402), a focusing lens (403), a first light splitting mirror (404), a forward light path device (405), a lateral light path device (406), a fluorescent path device (407);
The laser emission device (401), the optical fiber tube (402), the focusing lens (403), the laser detection chamber (372) and the first light splitting reflector (404) are arranged in an optical path, the focusing lens (403) is arranged on one side of the laser detection chamber (372), the forward light path device (405) is arranged on the opposite side of the laser detection chamber (372), the first light splitting reflector (404) is arranged on the other side of the laser detection chamber (372), and the lateral light path device (406) and the first light splitting reflector (404) are arranged at an angle;
the laser 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 onto each sperm cell flowing in the laser detection chamber (372);
the focused laser light, upon illuminating each of said sperm cells, forms a shadow behind said sperm cell due to its different size, thereby forming a forward optical signal, which is received by said forward optical access means (405); after the laser irradiates each sperm cell, lateral refraction light with different light intensities is formed on the sperm cell lateral direction due to different cell densities, so that a lateral light signal is formed, the lateral light signal is received by the lateral light path device (406) after being refracted by the first light splitting reflector (404), and the lateral light signal is an optical signal lower than Anm;
After the focused laser irradiates the fluorescence of each sperm cell, different fluorescent signals can be reflected, and after the laser irradiates the fluorescent probes on each sperm cell, the different fluorescent probes can be excited by the laser with different characteristic wavelengths, and then the fluorescent signals with different characteristic wavelengths can be emitted;
forming a first fluorescence signal after the fluorescence signal passes through the first light splitting reflector (404), wherein the first fluorescence signal is received by the fluorescence channel device (407); the first fluorescent signal is the fluorescent signal exceeding Anm.
7. The fully automated sperm cell tester as described in claim 6, wherein said fluorescence pathway apparatus (407) comprises a second light splitting mirror (471), a first fluorescence receiving apparatus (472), a third light splitting mirror (473), a second fluorescence receiving apparatus (474), and a third fluorescence receiving apparatus (475);
the first fluorescent signal irradiates the second light-splitting reflector (471) to form a second fluorescent signal after passing through the second light-splitting reflector, and the second fluorescent signal is received by the first fluorescent receiving device (472); the first fluorescent signal irradiates the second light-splitting reflector (471) and is refracted by the second light-splitting reflector to form a third fluorescent signal, and the third fluorescent signal is received by the third light-splitting reflector (473); the second fluorescent signal is the first fluorescent signal below Bnm, and the third fluorescent signal is the first fluorescent signal above Bnm;
The third fluorescent signal irradiates the third light-splitting reflector (473) and is refracted by the third light-splitting reflector to form a fourth fluorescent signal, and the fourth fluorescent signal is received by the second fluorescent receiving device (474); the third fluorescent signal irradiates the third light-splitting reflector (473) to form a fifth fluorescent signal after passing through the third light-splitting reflector, and the fifth fluorescent signal is received by the third fluorescent receiving device (475); the fourth fluorescent signal is the third fluorescent signal below Cnm and the fifth fluorescent signal is the third fluorescent signal above Cnm.
8. The fully automated sperm cell tester as described in claim 7, wherein said forward light path means (405) is a forward light bandpass filter and said lateral light path means (406) is a lateral light bandpass filter;
the numerical range of A is 490-510nm, the numerical range of B is 550-600nm, and the numerical range of C is 600-640nm;
the first fluorescence receiving device (472) is a first fluorescence band-pass filter, and the first fluorescence band-pass filter can pass the second fluorescence signal of 500-550 nm;
the second fluorescence receiving device (474) is a second fluorescence band-pass filter, and the second fluorescence band-pass filter can pass the fourth fluorescence signal of 550nm-600 nm;
The third fluorescence receiving device (475) is a third fluorescence band-pass filter, and the third fluorescence band-pass filter can pass the fifth fluorescence signal of 600nm-680 nm.
9. The fully automatic sperm cell tester as described in claim 1, wherein said control system (60) comprises control means (601) for controlling, an analog-to-digital converter (602), data analysis means (603), data storage means (604) and data output means (605) connected to said control means (601);
the photoelectric detection system (50) respectively converts the received optical signals into electric signals and sends the electric signals to the analog-to-digital converter (602), the analog-to-digital converter (602) converts the electric signals into digital signals and stores the digital signals in the data storage device (604), the data analysis device (603) decodes the digital signals into corresponding numerical values of various parameters for statistical analysis and generates analysis results, and the data output device (605) is used for outputting the analysis results.
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| CN201811599058.7A CN109612911B (en) | 2018-12-26 | 2018-12-26 | Full-automatic sperm cell detector |
| PCT/CN2019/073863 WO2020133635A1 (en) | 2018-12-26 | 2019-01-30 | Fully automatic sperm cell detector |
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| CN111323361B (en) * | 2020-03-17 | 2022-05-10 | 南通大学 | Method for quickly separating sperm head, sperm tail and normal viable sperm |
| CN111504869B (en) * | 2020-05-15 | 2021-06-08 | 中国计量科学研究院 | Flow type aggregate impurity analyzer |
| CN114527041A (en) * | 2020-10-30 | 2022-05-24 | 深圳市瑞图生物技术有限公司 | Display control method and device, sperm quality analysis system and storage medium |
| CN112698024B (en) * | 2020-12-08 | 2022-04-26 | 华中农业大学 | Immunoassay method based on differential impedance particle counting |
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| CN113092430B (en) * | 2021-04-09 | 2022-06-28 | 青岛复诺生物医疗有限公司 | Sperm active oxygen content measuring device and measuring method thereof |
| CN113156146A (en) * | 2021-04-12 | 2021-07-23 | 上海禹视科技有限公司 | Full-automatic high-throughput cell analysis equipment and detection method thereof |
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