A fully automatic detection system and method for milk somatic cells based on fluorescence analysis

Abstract

The present invention discloses a fully automatic detection system and method for milk somatic cells based on fluorescence analysis. The present invention completes automatic sampling and automatic mixing through an automatic sampling module, completes automatic focusing of stained samples through an automatic focusing module, and completes automatic photographing and automatic counting through an automatic photographing and counting module, thereby realizing fully automatic fluorescence microscopy. The present invention significantly improves the level of automated detection, ensures the consistency of the detection process, reduces manual operation errors, and thus improves experimental efficiency, data accuracy and reproducibility; compared with traditional offline detection or laboratory detection, the method and system proposed by the present invention have better real-time performance and accuracy, and also have a higher degree of automation.

Description

Technical Field

[0001] The invention belongs to the technical field of analysis and detection and is a method for detecting milk somatic cells, and in particular relates to a full-automatic detection system and method for milk somatic cells based on fluorescence analysis. Background Art

[0002] Somatic cell count can reflect the quality of milk production and the health of dairy cows, so the industry uses somatic cell count of milk as a quality standard. Currently, most tests in this area are conducted in offline laboratories. Common methods for milk somatic cell detection technology can be divided into indirect and direct methods.

[0003] Indirect methods infer the total somatic cell count by detecting changes in the physical and chemical properties of milk, such as the California Mastitis Test (CMT), the Wisconsin Mastitis Test (WMT), and the viscosity method. These methods use surfactants to release DNA in cells, which reacts with reagents to form gels, and the degree of gel is observed to determine inflammation. However, these methods have certain limitations due to problems such as untypical physical and chemical characteristics, strong subjectivity, and reliance on experience. In particular, they are difficult to be accurate when screening for subclinical inflammation, but they are still feasible as a preliminary screening method.

[0004] Among the direct methods, the classic methods include making smears and counting them under a microscope, and fluorescence microscopy, which uses fluorescent dyes to mark and detect under a fluorescence microscope. However, these methods are gradually replaced by flow cytometry due to their complex operation, dependence on manual labor, and low degree of automation. Flow cytometry uses a laser beam to illuminate a cell population suspended in a liquid, stimulating the DNA and RNA dyes in the cells to emit fluorescent signals. The pre-fluorescently stained cells are wrapped and diluted with sheath fluid, and passed through the optical detection system one by one to achieve accurate counting. This technology has the advantages of high sensitivity and high accuracy, but the operation process is complicated, the equipment and use costs are high, and cell morphology cannot be observed.

[0005] In summary, the current detection methods for milk somatic cells are mostly offline operations, which have the problem of poor real-time performance, more specifically, detection delays and lack of timeliness. In addition, the sample pre-treatment process is usually more complicated and requires a lot of manual intervention, affecting the overall efficiency. The data from offline detection is difficult to integrate with the data from other online monitoring systems in real time, which hinders the comprehensive analysis and decision-making of the data. The data interface and synchronization issues of offline detection are also more complicated, which increases the difficulty of system integration and has a low level of automation and intelligence.

[0006] Since online testing generates a large amount of data, especially some high-frequency and complex data, and integrating the testing equipment with the production line usually requires highly customized solutions, complex debugging and design, and is difficult to implement. Therefore, it is necessary to propose an online testing method and system for milk somatic cell testing. Summary of the invention

[0007] The present invention aims to solve the problems of low efficiency of traditional offline detection and low automation level of fluorescence analysis methods, and proposes a fully automatic detection system and method for milk somatic cells based on fluorescence analysis. The present invention realizes fully automatic fluorescence microscopy through an automatic sampling module, an automatic focusing module, an automatic photo counting module and a control module, which significantly improves the level of automated detection, ensures the consistency of the detection process, reduces manual operation errors, and thus improves experimental efficiency, data accuracy and reproducibility; compared with traditional offline detection or laboratory detection, it has good real-time performance, accuracy and high automation.

[0008] The technical solution adopted by the present invention is:

[0009] 1. A fully automatic detection system for milk somatic cells based on fluorescence analysis

[0010] An automatic sampling module is used to automatically extract a preset amount of sample and dye, and automatically mix the sample and dye to obtain a dyed sample, and then transport the dyed sample to the microfluidic chip;

[0011] An automatic focusing module is used to automatically adjust the position of the stained sample according to the imaging quality of the stained sample in the microfluidic chip, so as to automatically focus on the sample;

[0012] An automatic camera counting module is used to collect the stained sample image in the microfluidic chip and count the somatic cells in the stained sample image, thereby obtaining the detection result of the somatic cells;

[0013] The control module is used to obtain and send control signals to the automatic sampling module, the automatic focusing module and the automatic photo counting module, and upload the detection results of somatic cells to the terminal.

[0014] The automatic sampling module also includes an automatic cleaning pipeline and an automatic emptying pipeline.

[0015] The automatic focusing module includes an image processing module, a fluorescence microscope system, an electric stage, a stepper motor assembly and a stage driver, the control module is connected to the stepper motor assembly, the microfluidic chip is installed in the electric stage, the electric stage is connected to the stage driver, the stepper motor is connected to the stage driver, the fluorescence microscope system is installed above the electric stage, and the stepper motor assembly and the fluorescence microscope system are both connected to the image processing module; when the stained sample enters the microfluidic chip, the control module starts the stepper motor assembly, the stepper motor assembly drives the electric stage to move up and down within a preset moving range with a preset step length, and each moving position is recorded as a focus position, the fluorescence microscope system collects the stained sample image at each focus position and sends the stained sample image corresponding to each focus position to the image processing module, the image processing module generates an optimal focus position after judging the imaging quality of each stained sample image, and sends the optimal focus position to the stepper motor assembly, thereby controlling the electric stage to be located at the optimal focus position.

[0016] After focusing is completed, in the automatic photographing and counting module, the camera is started to automatically photograph the stained sample in the microfluidic chip to obtain the stained sample image, and then the image processing method is used to count the somatic cells of the stained sample images taken at different times, thereby obtaining the somatic cell detection result.

[0017] In the image processing module, after edge detection is performed on the dyed sample image corresponding to each focus position using the Sobel operator, the corresponding sample edge image is obtained, and then the average grayscale value of the sample edge image at the current focus position is calculated; the dyed sample images at different focus positions are traversed and processed to obtain the average grayscale values ​​of different focus positions, and the focus position with the highest average grayscale value is taken as the optimal focus position.

[0018] The electric object displacement stage is equipped with an LED light source, the power of the LED light source is 5W, and the rated current is 300mA.

[0019] A fishbone structure is arranged at the flow channel inlet of the microfluidic chip.

[0020] 2. A detection method of a fully automatic detection system for milk somatic cells based on fluorescence analysis

[0021] 1) Place the sample to be tested in the sample pool, the host computer sends a start command and controls the automatic sampling module to extract liquid from the sample pool and the dye pool into the mixing pool and complete the mixing of the system to obtain the dyed sample, and then inject the dyed sample into the inlet of the microfluidic chip flow channel;

[0022] 2) driving the electric stage with the microfluidic chip in the autofocus module so that the electric stage and the objective lens of the fluorescence microscope move relative to each other to achieve automatic focusing of the sample;

[0023] 3) The LED light source emits excitation light and illuminates the liquid in the flow channel of the microfluidic chip. The camera on the fluorescence microscope system collects real-time images and transmits the real-time collected stained sample images to the host computer. The collected stained sample images are processed and counted to obtain the detection results of somatic cells;

[0024] 4) After the test is completed, the automatic sampling module is controlled to perform self-cleaning of the pipeline and the cleaning of the mixing pool.

[0025] The specific contents of 2) are:

[0026] When the stained sample enters the microfluidic chip, the control module starts the stepper motor assembly, and the stepper motor assembly drives the electric stage to move up and down within a preset moving range with a preset step length, and each moving position is recorded as a focus position. The fluorescence microscope system collects the stained sample image at each focus position and sends the stained sample image corresponding to each focus position to the image processing module. The image processing module judges the imaging quality of each stained sample image and then generates the optimal focus position, and sends the optimal focus position to the stepper motor assembly, thereby controlling the electric stage to be located at the optimal focus position.

[0027] In the image processing module, after edge detection is performed on the dyed sample image corresponding to each focus position using the Sobel operator, the corresponding sample edge image is obtained, and then the average grayscale value of the sample edge image at the current focus position is calculated; the dyed sample images at different focus positions are traversed and processed to obtain the average grayscale values ​​of different focus positions, and the focus position with the highest average grayscale value is taken as the optimal focus position.

[0028] The beneficial effects of the present invention are:

[0029] Compared with the traditional laboratory offline detection method, the fully automatic detection system and method of milk somatic cells based on fluorescence analysis provided by the present invention simplifies the cumbersome operations such as sample dilution, staining and chip injection.

[0030] The present invention communicates with the detection system through the host computer, and each module can automatically execute the complete detection process, which significantly improves the automation level, reduces manual intervention, and improves overall efficiency without increasing costs. The present invention can perform fluorescence detection on milk somatic cells, which is of great significance to the quality evaluation of milk. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 It is a schematic diagram of the system framework of the present invention.

[0032] Figure 2 Schematic diagram of a control framework of an embodiment of the present invention.

[0033] Figure 3 This is a simplified schematic diagram of the automatic sampling module.

[0034] Figure 4 The figure is a flow chart of a method according to an embodiment of the present invention.

[0035] Figure 5 This is a flowchart of the autofocus module. DETAILED DESCRIPTION

[0036] The present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0037] The present invention proposes a fully automatic detection system for milk somatic cells based on fluorescence analysis. The system is a modular system that can automatically perform sampling, vibration mixing, focusing, photography and counting. Figure 1 and Figure 2 As shown, the system includes:

[0038] The automatic sampling module is used to automatically extract preset amounts of samples and dyes, automatically mix the samples and dyes to obtain dyed samples, and then transport the dyed samples to the microfluidic chip; the automatic sampling module also includes automatic cleaning pipelines and automatic emptying pipelines.

[0039] An automatic focusing module is used to automatically adjust the position of the stained sample according to the imaging quality of the stained sample in the microfluidic chip, so as to automatically focus on the sample;

[0040] The automatic camera counting module is used to collect the stained sample image in the microfluidic chip and count the somatic cells in the stained sample image, so as to obtain the detection result of the somatic cells.

[0041] The control module is used to obtain control signals from the terminal and send them to the automatic sampling module, the automatic focusing module and the automatic photo counting module, and upload the detection results of somatic cells to the terminal.

[0042] In this embodiment, the terminal is a host computer, which includes a graphical user interface, and the control module is a single-chip microcomputer. The host computer sends commands to the single-chip microcomputer of the system to control the start and stop of the automatic injection and focusing process, and at the same time, the microscope image is obtained through the camera, and the corresponding results are given in real time in the graphical user interface of the host computer through the image processing method. The functions and implementation methods of the automatic injection module, the automatic focusing module and the automatic photo counting module are described in detail below.

[0043] The automatic injection module is a key component of the system. The main body is a piping system that uses a pump to drive the circulation of liquid. Specifically, the host computer accurately controls the speed of the peristaltic pump by adjusting the PWM (pulse width modulation) duty cycle, thereby achieving accurate and repeatable liquid delivery. The host computer controls the vibration motor to automatically vibrate and mix through communication. The piping system is mainly based on a peristaltic pump, which has strong self-priming ability and a non-return function to ensure the unidirectional flow of the liquid. The automatic injection module has the characteristics of high accuracy and high automation, and can automatically complete the entire process after receiving the signal. In the event of a fault, the automatic injection module integrates a multi-channel control interface, which can be remotely controlled or manually controlled to start and stop, increasing the reliability and operational flexibility of the system.

[0044] The detection area is the observation room in the flow channel of the microfluidic chip. Samples are placed in the flow channel of the microfluidic chip. When somatic cells pass through the common microfluidic flow channel, accumulation, agglomeration, and clustering may occur occasionally at the entrance. In order to avoid this problem, the flow channel entrance has been improved. A fishbone structure is set at the flow channel entrance of the microfluidic chip, which can effectively separate the passing somatic cells and allow the somatic cells to pass through the flow channel evenly, which is convenient for subsequent photo analysis.

[0045] The autofocus module includes an image processing module, a fluorescence microscope system, an electric stage, a stepper motor assembly and a stage driver. The control module is connected to the microcontroller of the stepper motor assembly, the microfluidic chip is installed in the electric stage, the electric stage is connected to the stage driver, the stepper motor is connected to the stage driver, and the fluorescence microscope system is installed above the electric stage. The stepper motor assembly includes a stepper motor and a microcontroller connected to the stepper motor. The fluorescence microscope system includes a fluorescence microscope installed above the electric stage and a camera installed in the fluorescence microscope. The microcontroller of the stepper motor assembly and the camera of the fluorescence microscope system are both connected to the image processing module. An LED light source is installed on the electric stage, and the system has high brightness and miniaturization requirements for the light source. The power of the LED light source is 5W, the rated current is 300mA, and it is equipped with a driver chip and a shell. The stage driver is a ball screw, and the ball screw is driven by a stepper motor to achieve precise movement of the electric stage. The stage and the connected Z-axis translation stage can be controlled by the microcontroller in the stepper motor assembly by adjusting the step angle, speed, acceleration and other parameters of the stepper motor, so as to flexibly drive the stepper motor to drive the ball screw to move slightly and achieve precise focusing of the sample. After the sample is mixed in the automatic injection module, it is injected into the channel entrance of the microfluidic chip through a peristaltic pump. After the stained sample enters the microfluidic chip, the automatic focusing module starts working, using the automatic focus control method to accurately adjust the sample position to ensure the best shooting effect.

[0046] The focusing process is highly automated, and fast and stable focusing effects are achieved through closed-loop precision control. Figure 5 As shown, the specific process is as follows:

[0047] When the stained sample enters the microfluidic chip, the control module starts the stepper motor assembly, which drives the electric stage to move up and down within the preset moving range with a preset step length, and records each moving position as the focus position. The preset step length and moving range are optimized according to the specific application and sample characteristics of the fluorescence microscope. The camera of the fluorescence microscope system collects the stained sample image at each focus position and sends the stained sample image corresponding to each focus position to the image processing module. The image processing module determines the imaging quality of each stained sample image and then generates the optimal focus position, and sends the optimal focus position to the stepper motor assembly, thereby controlling the electric stage to be located at the optimal focus position, completing the precise adjustment of the sample position, and ensuring the best shooting effect.

[0048] In the image processing module, after edge detection is performed on the stained sample image corresponding to each focus position using the Sobel operator, the corresponding sample edge image is obtained. The Sobel operator can efficiently detect the horizontal and vertical edges in the image, enhance the edge information of the image, and is used to extract the edge features in the image. Then, the average grayscale value of the sample edge image at the current focus position is calculated. The higher the average grayscale value, the better the clarity and focus quality of the image; the stained sample images at different focus positions are traversed and processed to obtain the average grayscale values ​​of different focus positions, and the average grayscale value of each focus position is stored in a data structure for subsequent comparison. Arrays or lists and other structures can be used to save the corresponding positions and their average grayscale values. The focus position with the highest average grayscale value is used as the optimal focus position, and the stepper motor is controlled to move quickly and accurately to the determined optimal position to achieve accurate focus. Through the image processing module proposed by the present invention, the system can accurately find the best focus point; the entire focusing process can be automated, reduce manual intervention, and improve efficiency; the entire focusing process can also be adjusted according to the characteristics of different samples and fluorescence microscopes to meet various microscopic imaging requirements.

[0049] After focusing is completed, in the automatic camera counting module, the camera on the fluorescence microscope system is started to automatically take pictures of the stained samples in the microfluidic chip to obtain the stained sample images, and then the image processing method is used to count the somatic cells of the stained sample images taken at different times, so as to obtain accurate somatic cell detection results. The automatic camera counting module ensures the high quality and high accuracy of each image. The graphical user interface of the host computer is simple and easy to use, and can display the captured images and processing results in real time, allowing the operator to monitor the system operation status at any time.

[0050] Each module of the overall system has a rigorous process, high degree of automation, and strong accuracy. All modules can be manually fine-tuned to meet specific experimental needs. This system is different from traditional laboratory testing or offline testing methods. It can achieve online, real-time, and accurate testing. Through automated sampling, mixing, focusing, and photography, the detection efficiency and accuracy are greatly improved, and human errors and operational complexity are reduced. The close cooperation between the modules and the efficient communication between the host computer and the single-chip microcomputer ensure the accuracy and stability of the system.

[0051] The present invention also proposes a fully automatic detection method for milk somatic cells based on fluorescence analysis, such as Figure 4 As shown, the detection method comprises the following steps:

[0052] 1) After the milk in the production line pipeline is pumped into the sample bottle for sampling, the sample to be tested is placed in the sample pool. The host computer sends a start command and controls the automatic sampling module to drive the peristaltic pump to extract a certain amount of liquid from the sample pool and the dye pool into the mixing pool and complete the mixing of the system (that is, control the motor at the mixing pool to vibrate) to obtain the dyed sample; then control the peristaltic pump to inject the dyed sample into the inlet of the microfluidic chip flow channel; the microfluidic inlet has a special structural design to improve the subsequent observation effect. In the whole process, cleaning and emptying steps are interspersed to ensure the cleanliness of the system and minimize concentration fluctuations.

[0053] 2) The controller drives the electric stage with the microfluidic chip in the autofocus module, so that the electric stage and the objective lens of the fluorescence microscope move slightly relative to each other. The stage moves through the combination of the stepper motor and the ball screw, and the automatic focusing of the sample is achieved through the software.

[0054] 3) The LED light source emits excitation light and irradiates the liquid in the flow channel of the microfluidic chip through the optical system. The somatic cells stained with special dyes can be observed from the fluorescence microscope; the camera on the fluorescence microscope system collects real-time images and transmits the real-time collected stained sample images to the graphical user interface of the host computer. The software processes and counts the collected stained sample images, obtains the detection results of somatic cells and outputs them for display. The counting results of the software can reflect the quality of milk.

[0055] 4) After all the detection and analysis steps are completed, since the samples and dyes used are soluble in water, the microcontroller controls the water pump of the automatic sampling module to extract clean water from the clean water bottle for self-cleaning of the pipeline and cleaning of the mixing pool, and finally pumps away the waste liquid.

[0056] The following is further described by an embodiment:

[0057] Figure 3It is a simple schematic diagram of the automatic sampling module, which includes a milk pipeline 1, a two-way valve 2, a clean water bottle 3, a sample bottle 4, a dye bottle 5, a waste liquid bottle 6, a mixing pool 7, a microfluidic chip (i.e., an observation chamber) 8, an optical system 9, a camera 10, a host computer 11, a three-way valve 12, a light source 13, and a first peristaltic pump 14. The sample bottle 4 is connected to the milk pipeline 1 through the two-way valve 2 and the first peristaltic pump 14, and the milk pipeline 1 is used to circulate milk; the sample bottle 4 is also connected to the mixing pool 7 through the second peristaltic pump, the clean water bottle 3 is connected to the mixing pool 7, the dye bottle 5 is connected to the mixing pool 7 through the third peristaltic pump, the mixing pool 7 is connected to the microfluidic chip 8 through the three-way valve 12, and the microfluidic chip 8 is also connected to the waste liquid bottle 6; the microfluidic chip 8 is also equipped with an optical system 9, a light source 13, and a camera 10, and the camera 10 is connected to the host computer 11.

[0058] The automatic focusing module includes an electric translation stage, a stage and a fluorescence microscope system. The automatic photo counting module is mainly realized by the camera and the host computer software. The system is controlled by an STM32 microcontroller.

[0059] The system is operated by driving a peristaltic pump with a stepper motor, and sending the samples and dyes in the sample bottle and dye bottle into the mixing pool and mixing them automatically at a certain ratio according to the speed set by the software. Then the samples are pumped into the microfluidic chip (observation chamber). The cell molecules in the observation chamber will emit fluorescence under the excitation of the LED excitation light source, and the focus will be automatically adjusted through the autofocus module. The fluorescence image observed by the fluorescence microscope is captured by a camera and sent to the host computer for processing. The counting method gives the counting results in real time.

[0060] More specifically, since the dye sample needs to be injected in a 1:4 ratio, the milk sample and dye need to be pre-filled in the pipeline before the mixing pool is injected to prevent the injection ratio from being affected by external factors such as the length of the pipeline. The specific implementation method is to drive the two pumps to rotate a predetermined number of circles before the formal pumping, the sample pool peristaltic pump rotates 26.5 circles, that is, 6400×24.5 pulses are generated, the dye pump rotates 22 circles, and the part of the sample or dye that leaks into the mixing pool at the pipe mouth is cleaned with clean water, and then the waste liquid is pumped away. After all is completed, the proportional mixing injection is started again.

[0061] In this embodiment, a travel electromagnet is provided ( Figure 3Not marked in the figure), the purpose is to energize the travel electromagnet coil to compress the pipeline after the stained sample is sent to the observation chamber. The purpose of this is to prevent the waste liquid from the subsequent pipeline from flowing back; secondly, the camera used to take the image is not a high-speed camera, and the cells need to be decelerated as soon as possible until they can be observed during injection. Compressing the pipeline can improve the deceleration effect. In order to further achieve the purpose of deceleration, when injecting the sample, after the pump head rotates forward and pumps it into the observation chamber, it will be reversed for a short distance, which is equivalent to giving the liquid a back-drawing force, which is conducive to its deceleration, thereby facilitating faster observation and achieving better results.

[0062] After the test is completed, in the subsequent cleaning process, the clean water pump and the waste liquid pump are connected to the mixing tank, and then through the three-way valve, the mixing tank and the observation room can be fully rinsed and the waste liquid can be pumped away. The subsequent cleaning of the sample and dye pipelines can be achieved by connecting one end to a clean water bottle and quickly pumping back the clean water. Since the sample and dye pipelines need to be filled in advance and there is always liquid, if they are cleaned directly, the residual water in the pipeline will affect the concentration, so clean water cleaning is not arranged separately when the system is working. It can be directly replaced, disassembled and cleaned after the test is completed.

[0063] The microcontroller communicates with the host computer through the serial port, and the baud rate is set to 115200.

[0064] The pipes used in the system are food-grade silicone tubes, which are easy to disassemble and clean, avoid dead corners and slow flow areas, maximize the uniformity of the transported milk sample ingredients, and prevent chemical reactions. Silicone tubes have good temperature resistance and can usually be used in a temperature range of -60℃ to 200℃, which helps control the temperature during transportation.

[0065] The peristaltic pump uses a six-roller pump head. Compared with a three-roller pump head or a four-roller pump head, the six-roller design increases the number of rollers, which can effectively reduce the pulsation and fluctuation of the fluid, and is very suitable for scenarios with high requirements for flow stability. The setting of more rollers also reduces the gap between the rollers, thereby reducing the possibility of liquid reflux and recoil, and improving pumping efficiency. At the same time, it reduces the pressure in the pipeline and reduces the wear and fatigue of the pipeline.

[0066] The peristaltic pump is driven by a stepper motor, and the parameters are set to 6400 pulses for one rotation of the rotor. The amount pumped by the pump head is fixed for each rotation of the rotor. Therefore, when controlling, you only need to adjust the number of pulses generated to control the number of rotor rotations, thereby controlling the amount of sample injection.

[0067] Vibration motors are installed on both sides of the mixing tank, and the speed of the internal rotor is as high as 22,000 rpm. Since the rotor is equipped with an eccentric block, the centrifugal force generated during high-speed rotation causes the entire motor to vibrate, ensuring that the sample and dye system are fully mixed in the mixing tank, thereby ensuring the dyeing effect.

[0068] Finally, it should be noted that the above embodiments and explanations are only used to illustrate the technical solution of the present invention rather than to limit it. Those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the spirit and scope disclosed in the technical solution of the present invention, which should be included in the scope of protection of the claims of the present invention.

Claims

1. A fully automatic milk somatic cell detection system based on fluorescence analysis, characterized in that: include: An automatic sampling module is used to automatically extract a preset amount of sample and dye using a peristaltic pump, and automatically mix the sample and dye to obtain a dyed sample, and then transport the dyed sample to the microfluidic chip; An automatic focusing module is used to automatically adjust the position of the stained sample according to the imaging quality of the stained sample in the microfluidic chip, so as to automatically focus on the sample; An automatic camera counting module is used to collect the stained sample image in the microfluidic chip and count the somatic cells in the stained sample image, thereby obtaining the detection result of the somatic cells; A control module, used to obtain and send control signals to the automatic sampling module, the automatic focusing module and the automatic photo counting module, and to upload the detection results of somatic cells to the terminal; The automatic focusing module includes an image processing module, a fluorescence microscope system, an electric stage, a stepper motor assembly and a stage driver, the control module is connected to the stepper motor assembly, the microfluidic chip is installed in the electric stage, the electric stage is connected to the stage driver, the stepper motor is connected to the stage driver, the fluorescence microscope system is installed above the electric stage, and the stepper motor assembly and the fluorescence microscope system are both connected to the image processing module; when the stained sample enters the microfluidic chip, the control module starts the stepper motor assembly, the stepper motor assembly drives the electric stage to move up and down within a preset moving range with a preset step length, and each moving position is recorded as a focus position, the fluorescence microscope system collects the stained sample image at each focus position and sends the stained sample image corresponding to each focus position to the image processing module, the image processing module generates an optimal focus position after judging the imaging quality of each stained sample image, and sends the optimal focus position to the stepper motor assembly, thereby controlling the electric stage to be located at the optimal focus position; In the image processing module, after edge detection is performed on the dyed sample image corresponding to each focus position using the Sobel operator, the corresponding sample edge image is obtained, and then the average gray value of the sample edge image at the current focus position is calculated; the dyed sample images at different focus positions are traversed and processed to obtain the average gray values ​​of different focus positions, and the focus position with the highest average gray value is taken as the optimal focus position; A fishbone structure is arranged at the flow channel inlet of the microfluidic chip.

2. The fully automatic detection system for milk somatic cells based on fluorescence analysis according to claim 1, characterized in that: The automatic sampling module also includes an automatic cleaning pipeline and an automatic emptying pipeline.

3. The fully automatic detection system for milk somatic cells based on fluorescence analysis according to claim 1, characterized in that: After focusing is completed, in the automatic photographing and counting module, the camera is started to automatically photograph the stained sample in the microfluidic chip to obtain the stained sample image, and then the image processing method is used to count the somatic cells of the stained sample images taken at different times, thereby obtaining the somatic cell detection result.

4. The fully automatic detection system for milk somatic cells based on fluorescence analysis according to claim 1, characterized in that: The electric object displacement stage is equipped with an LED light source, the power of the LED light source is 5W, and the rated current is 300mA.

5. A detection method for implementing a fully automatic detection system for milk somatic cells based on fluorescence analysis according to claim 1, characterized in that: The detection method comprises the following steps: 1) Place the sample to be tested in the sample pool, the host computer sends a start command and controls the peristaltic pump in the automatic sampling module to extract liquid from the sample pool and the dye pool into the mixing pool and complete the mixing of the system to obtain the dyed sample, and then inject the dyed sample into the inlet of the microfluidic chip flow channel; 2) driving the electric stage with the microfluidic chip in the autofocus module so that the electric stage and the objective lens of the fluorescence microscope move relative to each other to achieve automatic focusing of the sample; 3) The LED light source emits excitation light and illuminates the liquid in the flow channel of the microfluidic chip. The camera on the fluorescence microscope system collects real-time images and transmits the real-time collected stained sample images to the host computer. The collected stained sample images are processed and counted to obtain the detection results of somatic cells; 4) After the test is completed, the automatic sampling module is controlled to perform self-cleaning of the pipeline and the cleaning of the mixing pool; The specific contents of 2) are: When the stained sample enters the microfluidic chip, the control module starts the stepper motor assembly, and the stepper motor assembly drives the electric stage to move up and down within a preset moving range with a preset step length, and each moving position is recorded as a focus position. The fluorescence microscope system collects the stained sample image at each focus position and sends the stained sample image corresponding to each focus position to the image processing module. The image processing module determines the imaging quality of each stained sample image and then generates the optimal focus position, and sends the optimal focus position to the stepper motor assembly, thereby controlling the electric stage to be located at the optimal focus position; In the image processing module, after edge detection is performed on the dyed sample image corresponding to each focus position using the Sobel operator, the corresponding sample edge image is obtained, and then the average grayscale value of the sample edge image at the current focus position is calculated; the dyed sample images at different focus positions are traversed and processed to obtain the average grayscale values ​​of different focus positions, and the focus position with the highest average grayscale value is taken as the optimal focus position.

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