Phage concentration detection method and device based on fluorescent labeling and image recognition

Abstract

This invention relates to the field of phage concentration detection technology, specifically to a method and device for phage concentration detection based on fluorescent labeling and image recognition. The device includes a support component, a detection component bolted to one side of the top of the support component, and a housing bolted to the same side of the top of the support component. The housing and detection component are bolted together. This invention achieves continuous automated detection of multiple samples through the integrated design of the support component and the detection component, combined with a sealed housing and a rotatable support stage. The placement slots and storage areas on the support stage can simultaneously accommodate multiple samples, which are sequentially positioned by a rotary motor, reducing manual intervention. The housing creates a dark environment, effectively isolating ambient light interference and improving the accuracy and reliability of fluorescence imaging. Furthermore, the operating interface, consisting of rubber gloves, allows users to operate samples in a sealed environment, further ensuring the cleanliness and consistency of the detection process. This significantly improves the efficiency and accuracy of phage concentration detection, shortening a single detection cycle to less than 1.5 hours.

Description

Technical Field

[0001] This invention relates to the field of phage concentration detection technology, specifically to a phage concentration detection method and device based on fluorescent labeling and image recognition. Background Technology

[0002] Bacteriophages are a class of viruses that infect bacteria and have significant research and application value in medicine, biology, and food safety. For example, they are used to treat bacterial diseases (phage therapy) and as indicators for the detection of specific bacteria. Accurate and rapid determination of phage concentration (i.e., titer) is a fundamental and crucial step in these applications. Traditional methods for phage concentration detection primarily rely on the double-layer agar plate method. This method is cumbersome, time-consuming (usually requiring overnight incubation), and the results are easily affected by factors such as operator experience and agar thickness, resulting in poor reproducibility. Furthermore, this method depends on visual observation and manual counting of phage plaques, which is highly subjective and makes high-throughput and automated detection difficult.

[0003] For example, application number CN201922474829.6, with an authorization announcement date of 20210618, describes a device for rapid detection of environmental bacteriophages. This device uses a vacuum pump to pass air through a double-layer plate coated with sensitive bacteria, adsorbing bacteriophages in the air. After incubation, the bacteriophages are counted via plaques. Although this method achieves air sample collection, it still requires 2 hours of adsorption and more than 6 hours of incubation, and relies on manual observation and counting, resulting in limited detection efficiency and accuracy. For example, a method for detecting bacteriophages, with application number CN201410570832.7 and authorization announcement date of 20170111, includes the following steps: S100, preparation of culture medium: preparing a host culture medium suitable for the growth of bacteriophage host bacteria; S200, coating of bacterial suspension: coating the bacteriophage host bacteria suspension onto the host culture medium to form a detection culture medium; S300, detection of bacteriophages: including: detection of bacteriophages in gas: first exposing the detection culture medium to the area to be tested, then culturing, and calculating the number of bacteriophages in the gas using the Omeryansky formula; and detection of bacteriophages in liquid: first coating the liquid to be tested onto the detection culture medium, then culturing, and calculating the number of bacteriophages in the liquid. This method also suffers from problems such as long processing time, low throughput, and high subjectivity. In recent years, some methods based on optical principles have been proposed, such as the patent with application number CN202511573130.9, which uses the change in turbidity of the host bacterial suspension caused by bacteriophage infection to indirectly determine the phage. However, this method can only detect qualitatively, not quantitatively, the concentration of bacteriophages.

[0004] Traditional phage concentration detection techniques generally suffer from drawbacks such as cumbersome and time-consuming operations, reliance on overnight incubation, and high consumption of culture medium. Furthermore, result interpretation is highly dependent on manual observation and counting, leading to subjectivity and susceptibility to errors. These methods also have significant limitations in throughput and automation, making it difficult to achieve high-throughput, rapid batch analysis of multiple samples. They also lack integrated image acquisition and data analysis modules, hindering real-time, dynamic monitoring of the phage growth process for rapid result acquisition. In summary, current technologies generally lack a rapid, automated, high-throughput phage concentration detection solution that provides objective and accurate results. Summary of the Invention

[0005] The purpose of this invention is to provide a method and device for detecting phage concentration based on fluorescent labeling and image recognition, so as to overcome the above-mentioned shortcomings in the prior art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A phage concentration detection device based on fluorescent labeling and image recognition includes a carrier assembly. A detection component is bolted to one side of the top of the carrier assembly. A housing is bolted to one side of the top of the carrier assembly, and the housing and detection component are bolted together. A lighting monitoring module is embedded in the top of the housing. A cover plate is bolted to one side of the outer wall of the housing, and two cylinders are bolted to one side of the cover plate. A rubber glove is bolted to one end of each cylinder. A display screen is bolted to one side of the outer wall of the housing. A door is hinged to one side of the outer wall of the housing, and a touch screen is embedded in one side of the outer wall of the door. A door handle is bolted to one side of the outer wall of the door. The supporting component includes a base, a mounting seat is mounted on the top center of the base via a bearing, and a support platform is bolted to the top of the mounting seat. Four placement slots arranged in a circular array are formed on the top outer wall of the support platform, and lower lighting modules are bolted into each placement slot. A glass plate located above the lower lighting modules is bolted into each placement slot. A storage area is formed at the top center of the support platform, and a stepped test tube rack is bolted into the storage area. A drawer box is slidably inserted into the storage area. A rotary motor is bolted to the center of one side of the inner wall of the base, and the output end of the rotary motor is fixedly connected to the mounting seat via a coupling.

[0007] Furthermore, a power module and a control module are bolted to both sides of the inner wall of the base, and the control module is electrically connected to the power module and the rotary motor respectively through wires.

[0008] Furthermore, the detection component includes a support frame, which is bolted to one side of the top of the base, and an installation groove is provided on one side of the outer wall of the support frame.

[0009] Furthermore, a groove is provided on one side of the outer wall of the support frame, and two slide rods are bolted inside the groove.

[0010] Furthermore, a lifting screw is installed inside the slide groove via a bearing, and a mounting shell is threaded onto the outside of the lifting screw. The mounting shell is slidably mounted on the two slide rods.

[0011] Furthermore, a drive motor is bolted to one side of the inner wall of the mounting groove, and the output end of the drive motor is fixedly connected to one end of the lifting screw through a coupling.

[0012] Furthermore, a vision module and a computing communication module are bolted inside the mounting housing, and the computing communication module is electrically connected to the vision module, the drive motor, and the touch screen via wires.

[0013] Furthermore, two upper lighting modules are bolted to the outer wall of one side of the mounting housing, and the computing communication module is electrically connected to the upper lighting modules via wires.

[0014] Furthermore, the vision module includes a camera and an image processing unit, which is used to capture images of fluorescent markers and calculate phage concentration using an image recognition algorithm.

[0015] A phage concentration detection method based on fluorescent labeling and image recognition includes the following steps: S1. Sample preparation and labeling: The phage sample to be tested is mixed with a specific fluorescent dye and incubated to form a fluorescently labeled phage sample; S2. Sample loading and sealing: Place multiple marked samples in the placement slots or test tube racks on the support platform, then close the door to form a sealed dark box to block ambient light. If necessary, internal operations can be performed in a sealed environment through the operating interface consisting of a cover plate, cylinder and rubber gloves. The operation process will be displayed on the screen. S3. Parameter setting and initialization: Input the detection parameters through the touch screen, and the control module initializes the system accordingly to prepare for automatic detection; S4. Automatic positioning and focusing: The control module starts the rotary motor to rotate the support platform, moving the samples to the detection position in sequence. Then, the drive motor is started, and the height of the vision module is adjusted by the lifting screw to make it accurately focus on the current sample. S5. Fluorescence Excitation and Imaging: The control module activates the lower illumination module and / or the upper illumination module to excite sample fluorescence, while the illumination monitoring module ensures stable light intensity. The camera of the vision module then acquires clear fluorescence images in a closed environment. S6. Image Processing and Analysis: The computing and communication module processes the acquired images, including image noise reduction and background correction, using image recognition algorithms to identify and count fluorescent spots, and calculating phage concentration based on the number of spots, field of view area and sample volume, or performing concentration conversion based on a pre-stored standard curve. (Based on the original description that “the glass plate (17) of the placement slot (15) can also be placed in the test tube of the test tube rack (19),” there are two measurement methods, as shown below:) Slide method (static): For the "placement tank", the sample is prepared as an immobilized thin layer sample. For example, after mixing bacteriophage with fluorescent dye, it is dropped onto a glass slide, and perhaps a coverslip is also added to form a liquid thin layer. In this state, the flow of liquid and the Brownian motion of bacteriophage are greatly restricted to a two-dimensional plane, and the range of movement is very limited. Test tube method (dynamic / static): directly measuring liquid in a test tube, the random movement of bacteriophages is indeed a huge challenge; As described in the original content (the visual module focuses on the current sample and glass plate), this application uses the slide method. By using the "slide method" and letting it stand for a period of time, the bacteriophages settle onto the surface of the glass plate or are confined within a thin layer. Their movement is negligible within the exposure time (milliseconds), so they can be "frozen" by the camera into a relatively stationary point. This is a common strategy in many current microscope-based bacterial / virus counting methods, so there is no need to add a trajectory capture function.

[0016] Regarding fluorescent spots: Fluorescent spots are not the shape of the bacteriophage itself, but rather a "Airy spot" formed by the light emitted by the fluorescent dye on the surface of the bacteriophage after passing through the optical system. The size of this spot (2-5 μm) is determined by the wavelength of the light and the numerical aperture of the microscope objective, and is much larger than the bacteriophage itself. Pixel resolution (5 million) only divides the image into how many small grids. What truly determines whether "details can be seen" is the optical resolution (determined by the objective). Furthermore, the goal of this application is not to "see" the bacteriophage, but to "identify" and "count" that 2-5 μm fluorescent spot, which is a completely different technical requirement. Secondly, appropriate content has been added below, and effective recognition is ensured through the design of the objective lens.

[0017] S7. Result Output: The calculated phage concentration value is displayed on the touch screen, and data storage, export or transmission is supported through the computing communication module; S8. Inspection Completion and Reset: After the inspection cycle is completed, the equipment controls all execution components to reset. The user can open the cabinet door to clean the support platform and other components for the next use.

[0018] In the above technical solution, the phage concentration detection method and device based on fluorescent labeling and image recognition provided by the present invention have the following beneficial effects: This invention achieves continuous automated detection of multiple samples through the integrated design of the support and detection components, combined with a sealed enclosure and a rotatable stage. Compared to traditional methods that require overnight incubation (8-24 hours), this invention can complete the entire detection process from sample loading to result output within 1.5 hours. The placement slots and storage areas on the stage can simultaneously accommodate multiple samples, which are sequentially positioned by a rotary motor, reducing manual intervention. The enclosure creates a dark environment, effectively isolating ambient light interference and improving the accuracy and reliability of fluorescence imaging. Furthermore, the rubber glove-based operating interface allows users to handle samples in a sealed environment, further ensuring the cleanliness and consistency of the detection process and significantly improving the efficiency and accuracy of phage concentration detection.

[0019] This invention achieves automated and stable operation of the detection process through the coordinated control of the control module and the power supply module. The lifting screw and drive motor in the detection component allow the vision module to precisely adjust the height, ensuring rapid focusing on different samples. The combined use of the upper and lower illumination modules optimizes fluorescence excitation conditions, and the illumination monitoring module maintains stable light intensity, thereby obtaining high-quality images. The integration of the computing and communication module with the vision module enables real-time processing and analysis of image data, reducing human error and improving the repeatability and reliability of the detection results.

[0020] This invention employs an image recognition algorithm for counting fluorescent spots and calculating concentration, avoiding the subjectivity and cumbersome operation of traditional methods. The vision module includes a camera and an image processing unit, which can automatically identify fluorescently labeled bacteriophages and perform concentration conversion using a pre-stored standard curve, greatly shortening the detection time. At the same time, the detection method is standardized, making the operation simple and fast, suitable for rapid screening in laboratories and industrial sites, and improving the versatility and practicality of bacteriophage concentration detection.

[0021] The device of this invention has a compact structure and complete functions. It provides a user-friendly human-machine interface through a touch screen and a display screen, which facilitates parameter setting and result viewing. The data storage and export functions support subsequent analysis and traceability. In addition, the device is easy to clean and maintain, which extends its service life and reduces operating costs. It has broad application value in the field of biological detection. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this invention. For those skilled in the art, other drawings can be obtained based on these drawings.

[0023] Figure 1 This is a schematic diagram of the overall structure of the phage concentration detection method and device based on fluorescent labeling and image recognition of the present invention.

[0024] Figure 2 This is a schematic diagram of the carrier component and detection component structure provided in the embodiment of the phage concentration detection method and equipment based on fluorescent labeling and image recognition of the present invention.

[0025] Figure 3 This is a schematic diagram of the supporting component structure provided in the embodiment of the phage concentration detection method and equipment based on fluorescent labeling and image recognition of the present invention.

[0026] Figure 4 This is a top view of the supporting component provided in the embodiment of the phage concentration detection method and equipment based on fluorescent labeling and image recognition of the present invention.

[0027] Figure 5 This is a schematic diagram of the detection component structure provided in the embodiment of the phage concentration detection method and equipment based on fluorescent labeling and image recognition of the present invention.

[0028] Figure 6 This is a side view of the detection component provided in the embodiment of the phage concentration detection method and equipment based on fluorescent labeling and image recognition of the present invention.

[0029] Figure 7 This is a flowchart illustrating the method and equipment embodiment of the phage concentration detection method and device based on fluorescent labeling and image recognition of the present invention.

[0030] Explanation of reference numerals in the attached figures: 1. Load-bearing component; 2. Shell; 3. Door; 4. Touch screen; 5. Display screen; 6. Cover plate; 7. Cylinder; 8. Rubber glove; 9. Lighting and monitoring module; 10. Detection component; 11. Door handle; 12. Base; 13. Mounting base; 14. Support platform; 15. Placement slot; 16. Lower lighting module; 17. Glass plate; 18. Storage area; 19. Test tube rack; 20. Drawer box; 21. Control module; 22. Rotary motor; 23. Power module; 24. Support frame; 25. Mounting slot; 26. Slide rail; 27. Lifting screw; 28. Slide rod; 29. ​​Mounting shell; 30. Computing and communication module; 31. Vision module; 32. Drive motor; 33. Upper lighting module. Detailed Implementation

[0031] To enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.

[0032] like Figure 1-6As shown, the embodiment of the present invention includes a support component 1, a detection component 10 bolted to one side of the top of the support component 1, a housing 2 bolted to one side of the top of the support component 1, and the housing 2 and the detection component 10 bolted together. A lighting monitoring module 9 is bolted into the top of the housing 2. The lighting monitoring module 9 has a built-in light intensity sensor and adjustment circuit, which monitors the lighting intensity inside the housing in real time. It communicates with the control module 21 and automatically adjusts the brightness of the upper lighting module 33 and the lower lighting module 16 to maintain the stability of the fluorescence excitation conditions and improve image quality. A cover plate 6 is bolted into one side of the outer wall of the housing 2, and two cylinders 7 are bolted to one side of the cover plate 6. A rubber glove 8 is bolted to one end of each cylinder 7. The rubber gloves 8 allow operators to place or adjust samples in a closed environment without opening the box, avoiding sample contamination and ambient light leakage, and ensuring the cleanliness of the testing process. A display screen 5 is bolted to one side of the outer wall of the box shell 2, showing the operation process including the rubber gloves 8, facilitating observation of the interior of the box shell 2. A door 3 is hinged to one side of the outer wall of the box shell 2, and a touch screen 4 is embedded in one side of the outer wall of the door 3. A door handle 11 is bolted to one side of the outer wall of the door 3, used for manually opening and closing the door 3 for convenient maintenance and sample loading. The supporting assembly 1 includes a base 12, with a mounting seat 13 mounted at the top center of the base 12 via a bearing, and the top of the mounting seat 13 is bolted... The system is equipped with a support stage 14, the rotation of which is controlled by a control module 21 to ensure sequential sample switching. Four placement slots 15 arranged in a ring array are formed on the top outer wall of the support stage 14. These slots are used to place sample dishes. A lower illumination module 16 is bolted inside each placement slot 15. The lower illumination module 16 uses an LED or laser light source to generate excitation light of a specific wavelength, which penetrates a glass plate 17 to illuminate the sample, optimizing the fluorescence signal. Its brightness is adjusted by the control module 21 to ensure consistent illumination. A glass plate 17, made of transparent material, is bolted inside each placement slot 15 and located above the lower illumination module 16. This glass plate 17 supports the sample and allows excitation light and fluorescence transmission while preventing interference. Sample contamination or displacement; a storage area 18 is provided at the center of the top of the support platform 14, and a stepped test tube rack 19 is bolted inside the storage area 18. The stepped structure of the test tube rack 19 facilitates the placement of test tubes of different sizes; a drawer box 20 is slidably inserted inside the storage area 18, which is used to store auxiliary reagents for testing and can be slidably removed for easy access to miscellaneous items; a rotary motor 22 is bolted to the center of one side of the inner wall of the base 12. The rotary motor 22 is controlled by the control module 21 and drives the support platform 14 to rotate, so that each placement slot 15 is aligned with the detection component 10 in sequence, realizing continuous testing of multiple samples; and the output end of the rotary motor 22 is fixedly connected to the mounting base 13 through a coupling. In one embodiment provided by the present invention, such as Figure 4As shown, a power module 23 and a control module 21 are bolted to both sides of the inner wall of the base 12. The control module 21 has an embedded controller that coordinates the operation of the rotary motor 22, the lighting module, and the detection component 10. It receives instructions from the touch screen 4, executes preset detection programs, and processes sensor data. The power module 23 provides power to the entire device, including the motor, lighting module, and electronic module, ensuring stable operation. The control module 21 is electrically connected to the power module 23 and the rotary motor 22 via wires. In another embodiment provided by the present invention, such as Figure 5-6 As shown, the detection assembly 10 includes a support frame 24, which is bolted to one side of the top of the base 12. A mounting groove 25 and a sliding groove 26 are formed on the outer wall of one side of the support frame 24. Two sliding rods 28 are bolted inside the sliding groove 26, providing sliding support and preventing the mounting housing 29 from rotating. A lifting screw 27 is mounted inside the sliding groove 26 via bearings and driven by a drive motor 32. This screw drives the mounting housing 29 up and down through threaded transmission, precisely adjusting the height of the vision module 31 to accommodate different samples. The system meets the focusing requirements; and the lifting screw 27 is externally threaded with a mounting shell 29, which is slidably mounted on two slide rods 28. A drive motor 32 is bolted to one side of the inner wall of the mounting groove 25. The drive motor 32 is a stepper motor or servo motor, controlled by the computing communication module 30, which drives the lifting screw 27 to rotate, thus achieving automatic focusing; and the output end of the drive motor 32 is fixedly connected to one end of the lifting screw 27 via a coupling. Inside the mounting shell 29, a vision module 31 and a computing communication module 30 are bolted to respectively. 31 includes a high-resolution camera and an image processing unit. The camera captures images of fluorescently labeled samples, and the image processing unit runs image recognition algorithms such as spot counting and contour analysis to identify phage fluorescent spots and calculate concentration values ​​based on pre-stored standard curves. The computing communication module 30 is a core processor that receives image data from the vision module 31, executes concentration calculation algorithms, and controls the drive motor 32, the upper illumination module 33, and communication with the touch screen 4. It is also responsible for data storage and export. The computing communication module 30 is electrically connected to the vision module 31, the drive motor 32, and the touch screen 4 via wires. Two upper illumination modules 33 are bolted to the outer wall of one side of the mounting housing 29. The upper illumination modules 33 provide supplemental illumination from above and work in conjunction with the lower illumination module 16 to provide uniform fluorescence excitation conditions. Their switching and brightness are adjusted by the computing communication module 30. The computing communication module 30 is electrically connected to the upper illumination modules 33 via wires. The vision module 31 includes a camera and an image processing unit. The vision module 31 is used to capture images of fluorescently labeled samples and calculate phage concentrations using image recognition algorithms.

[0033] The carrier assembly 1 drives the carrier stage 14 to rotate via a rotary motor 22, allowing multiple samples (placed in the placement slot 15 or test tube rack 19) to move sequentially to the detection position for continuous detection. The detection assembly 10 adjusts the height of the vision module 31 via a lifting screw 27 and a drive motor 32 to ensure precise focusing on different samples. The airtight design of the enclosure 2, combined with the illumination monitoring module 9, effectively eliminates ambient light interference and improves fluorescence imaging quality. The operating interface, consisting of rubber gloves 8 and a cover plate 6, allows users to make internal adjustments during detection without opening the enclosure door, maintaining the stability of the darkroom environment. The touch screen 4 and display screen 5 provide intuitive human-computer interaction, facilitating parameter setting and result viewing.

[0034] The power module 23 uses a rechargeable lithium battery, supporting at least 4 hours of operation without an external power source, improving portability. The control module 21 uses an STM32 series microcontroller and communicates with the computing communication module 30 via a CAN bus, achieving more efficient motion control and data synchronization. The support frame 24 of the detection component 10 is made of aluminum alloy, reducing weight while ensuring structural strength; a cooling fan is also added inside the mounting slot 25 to prevent the drive motor 32 from overheating. The camera resolution of the vision module 31 is increased to 5 megapixels, and it incorporates an automatic white balance function to adapt to the spectral characteristics of different fluorescent dyes. The vision module 31 is also equipped with an objective lens of appropriate magnification (e.g., a 4x or 10x objective lens), so that the "pixel equivalent" of its image on the 5-megapixel camera reaches approximately 0.5-1 μm / pixel. Therefore, a fluorescent spot with a diameter of 5 μm can occupy 25-100 pixels, which is more than enough for any image recognition algorithm to perform stable and accurate recognition and counting.

[0035] The lifting screw 27 uses a ball screw, which, together with the linear guide rail of the slide bar 28, enables the lifting accuracy of the mounting housing 29 to reach ±0.1mm. The drive motor 32 uses a stepper motor, and the control module 21 ensures rapid focusing of the vision module 31 through a closed-loop control algorithm. The image processing unit of the vision module 31 integrates deep learning algorithms (such as convolutional neural networks), which can identify fluorescent spots in real time and filter non-specific signals, improving counting accuracy. The computing and communication module 30 supports 4G network and cloud platform connectivity, enabling remote transmission and storage of detection data, facilitating data sharing and analysis across multiple locations.

[0036] like Figure 7 The phage concentration detection method based on fluorescent labeling and image recognition, as shown, includes the following steps: S1. Sample preparation and labeling: The phage sample to be tested is mixed with a specific fluorescent dye and incubated to form a fluorescently labeled phage sample; S2. Sample loading and sealing: Place multiple marked samples in the placement slot 15 of the support platform 14 or the test tube rack 19, then close the box door 3 to form a sealed dark box in the box shell 2 to isolate ambient light. If necessary, internal operations can be carried out in a sealed environment through the operation interface composed of the cover plate 6, the cylinder 7 and the rubber glove 8, and the operation process will be displayed on the display screen 5. S3. Parameter setting and initialization: Input the detection parameters through the touch screen 4, and the control module 21 initializes the system accordingly to prepare for automatic detection; S4. Automatic positioning and focusing: The control module 21 starts the rotary motor 22 to rotate the support stage 14, moving the samples to the detection position in sequence. Then, the drive motor 32 is started, and the height of the vision module 31 is adjusted by the lifting screw 27 to make it accurately focus on the current sample. S5. Fluorescence excitation and imaging: Control module 21 turns on lower illumination module 16 and / or upper illumination module 33 to excite sample fluorescence, while illumination monitoring module 9 ensures stable light intensity, and the camera of vision module 31 then acquires clear fluorescence images in a closed environment. S6. Image Processing and Analysis: The computing and communication module 30 processes the acquired images, including image noise reduction and background correction, using image recognition algorithms to identify and count fluorescent spots, and calculating the phage concentration based on the number of spots, field of view area and sample volume, or performing concentration conversion based on a pre-stored standard curve. S7. Result output: The calculated phage concentration value is displayed on the touch screen 4, and data storage, export or transmission is supported through the computing communication module 30; S8. Inspection Completion and Reset: After the inspection cycle is completed, the equipment controls all execution components to reset. The user can open the door 3 to clean the support platform 14 and other components for the next use.

[0037] The upper illumination module 33 and lower illumination module 16 employ multi-band LED light sources (such as ultraviolet, blue, and green light), which can switch excitation wavelengths according to different fluorescent dyes, improving detection flexibility. The camera in the vision module 31 is equipped with a motorized zoom lens, and combined with the real-time image processing of the computing and communication module 30, it can adapt to different sample thicknesses and concentration ranges. The image processing unit integrates a machine learning model, which, trained on historical data, can automatically optimize recognition parameters and reduce false positive results.

[0038] The number of placement slots 15 on the support stage 14 can be increased to eight, and a replaceable design allows users to change the dedicated clamps according to the sample type. The stepped structure of the test tube rack 19 is optimized to be adjustable, supporting the accommodation of test tubes from 1.5mL centrifuge tubes to 15mL tubes. The vision module 31 of the detection component 10 adopts a dual-camera system (one for wide-angle scanning and one for detailed imaging), combined with multi-source illumination, to achieve faster image acquisition and higher accuracy spot counting. In addition, the device also integrates environmental sensors (temperature and humidity), and the control module 21 can automatically adjust the detection parameters according to environmental data to ensure the reliability of the results.

[0039] It should be noted that: The specific details of the phage concentration detection method are as follows: S1. Sample preparation and fluorescent labeling: The phage sample to be tested is mixed with a specific fluorescent dye and incubated to allow the phage to bind to the fluorescent dye, forming a fluorescently labeled phage sample. S2. Sample loading and preparation in a closed environment: Open the door 3 and place multiple fluorescently labeled phage samples into the placement slot 15 or test tube rack 19 of the support platform 14 of the detection equipment; close the door 3 to create a sealed dark environment in the shell 2 to isolate external light interference; during the detection process, the user can use the sealed operation interface on the shell 2, which consists of a cover plate 6, a cylinder 7 and a rubber glove 8, to take out or place or fine-tune the internal samples or tools in the drawer box 20 without damaging the sealed environment, and the operation process will be displayed on the display screen 5; S3. Device Initialization and Parameter Settings: The device is started by using the touch screen 4 as the input interface, and the detection parameters are set, including lighting intensity, image acquisition parameters, platform rotation speed and image recognition algorithm threshold. S4. Automated Sample Positioning and Visual Focusing: According to the set parameters, the control module 21 starts the rotary motor 22 to drive the carrier platform 14 to rotate, so that each sample moves to the detection position in sequence; then, the control module 21 starts the drive motor 32, and adjusts the height of the mounting shell 29 through the lifting screw 27, so as to drive the vision module 31 and the upper illumination module 33 inside it to rise and fall until the vision module 31 is precisely focused on the current sample. S5. Fluorescence Excitation and Image Acquisition: The control module 21 activates the lower illumination module 16 and / or the upper illumination module 33 to illuminate the sample from below and / or above to excite the fluorescent marker to produce fluorescence; during this process, the illumination monitoring module 9 monitors the illumination status to ensure stable light intensity; subsequently, the camera of the vision module 31 captures high-quality fluorescent images of the sample inside the sealed enclosure 2. S6. Image Processing and Concentration Calculation: The vision module 31 or the computing and communication module 30 processes the acquired fluorescence image by performing the following sub-steps: S6.1: Image preprocessing: Denoising, contrast enhancement, and background correction are performed on the original image to highlight the fluorescence signal; S6.2: Fluorescent Spot Recognition and Counting: Using image recognition algorithms based on machine learning models, such as convolutional neural networks, fluorescent spots in images can be identified and segmented, and their number can be counted. At the same time, the fluorescence intensity, area, and roundness features of the spots can be extracted to help determine the phage status or distinguish non-specific signals. S6.3: Concentration Conversion: Based on the number of identified fluorescent spots, combined with the actual sample area and sample volume corresponding to the field of view, the phage concentration is calculated using the formula C=N / S*V, where C is the concentration, N is the number of fluorescent spots, S is the area, and V is the volume; or the fluorescence signal characteristics are converted into phage concentration values ​​based on a standard curve established in advance using samples with known concentrations. S7. Results Output and Display: The computing and communication module 30 sends the final calculated phage concentration data to the touch screen 4, which serves as the output interface, for clear display. It can also be viewed interactively on the touch screen 4 and supports data export, storage, or remote transmission. S8. Equipment reset and cleaning: After the test is completed, the equipment automatically controls the reset of each component, opens the door 3, and cleans the support platform 14, the placement slot 15 and the glass plate 17 for the next use.

[0040] The foregoing has only described certain exemplary embodiments of the present invention by way of illustration. Undoubtedly, those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the foregoing drawings and descriptions are illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.

Claims

1. A device for detecting the concentration of bacteriophages based on fluorescent labeling and image recognition, comprising a support assembly (1), characterized in that: The top side of the bearing assembly (1) is bolted with a detection assembly (10), and the top side of the bearing assembly (1) is bolted with a housing (2), and the housing (2) and the detection assembly (10) are bolted together. The top of the housing (2) is bolted with a lighting monitoring module (9), and the outer wall of the housing (2) is bolted with a cover plate (6), and the cover plate (6) is bolted with two cylinders (7). One end of the cylinder (7) is bolted with a rubber glove (8), and the outer wall of the housing (2) is bolted with a display screen (5). The outer wall of the housing (2) is bolted with a door (3) via a hinge, and the outer wall of the door (3) is bolted with a touch screen (4). The outer wall of the door (3) is bolted with a door handle (11). The bearing assembly (1) includes a base (12), and the top of the base (12) is bolted with a door handle (11). A mounting base (13) is installed at the center of the base via a bearing, and a support platform (14) is bolted to the top of the mounting base (13). Four placement slots (15) arranged in a ring array are opened on the top outer wall of the support platform (14), and a lower lighting module (16) is bolted inside the placement slot (15). A glass plate (17) located above the lower lighting module (16) is bolted inside the placement slot (15). A storage area (18) is opened at the center of the top of the support platform (14), and a stepped test tube rack (19) is bolted inside the storage area (18). A drawer box (20) is slidably inserted inside the storage area (18). A rotary motor (22) is bolted to the center of one side of the inner wall of the base (12), and the output end of the rotary motor (22) is fixedly connected to the mounting base (13) via a coupling.

2. The fluorescent marker and image recognition based phage concentration detection apparatus according to claim 1, characterized in that, The power module (23) and control module (21) are bolted to both sides of the inner wall of the base (12), and the control module (21) is electrically connected to the power module (23) and the rotary motor (22) through wires.

3. The fluorescent marker and image recognition based phage concentration detection apparatus according to claim 1, characterized in that, The detection component (10) includes a support frame (24), which is bolted to one side of the top of the base (12), and an installation groove (25) is provided on the outer wall of one side of the support frame (24).

4. The fluorescent marker and image recognition based phage concentration detection apparatus according to claim 3, characterized in that, The support frame (24) has a groove (26) on one side of its outer wall, and two slide rods (28) are bolted inside the groove (26).

5. The phage concentration detection device based on fluorescent labeling and image recognition according to claim 4, characterized in that, The sliding groove (26) is equipped with a lifting screw (27) through a bearing, and the lifting screw (27) is threaded with a mounting shell (29). The mounting shell (29) is slidably mounted on two sliding rods (28).

6. The phage concentration detection device based on fluorescent labeling and image recognition according to claim 5, characterized in that, A drive motor (32) is bolted to one side of the inner wall of the mounting groove (25), and the output end of the drive motor (32) is fixedly connected to one end of the lifting screw (27) through a coupling.

7. The phage concentration detection device based on fluorescent labeling and image recognition according to claim 6, characterized in that, The mounting housing (29) is bolted with a vision module (31) and a computing communication module (30), and the computing communication module (30) is electrically connected to the vision module (31), the drive motor (32) and the touch screen (4) through wires.

8. The phage concentration detection device based on fluorescent labeling and image recognition according to claim 7, characterized in that, Two upper lighting modules (33) are bolted to one side of the outer wall of the mounting housing (29), and the computing communication module (30) is electrically connected to the upper lighting module (33) via a wire.

9. The phage concentration detection device based on fluorescent labeling and image recognition according to claim 7, characterized in that, The vision module (31) includes a camera and an image processing unit. The vision module (31) is used to capture images of fluorescent markers and calculate the phage concentration using an image recognition algorithm.

10. A method for phage concentration detection based on fluorescent labeling and image recognition, characterized in that, Includes the following steps: S1. Sample preparation and labeling: Phage samples are mixed with fluorescent dyes and incubated for labeling; S2. Sample loading and sealing: Mark the sample and place it on the support platform (14), close the box door (3) to form a dark box, and operate it through the operation interface if necessary, and display it on the display screen (5); S3. Parameter setting and initialization: Input parameters on the touch screen (4), and initialize the system in the control module (21); S4. Automatic positioning and focusing: The rotary motor (22) moves the sample to the detection position and drives the motor (32) to adjust the height of the vision module (31) for focusing; S5. Fluorescence excitation and imaging: The lower illumination module (16) and / or the upper illumination module (33) are turned on to excite fluorescence, and the vision module (31) acquires the image; S6. Image Processing and Analysis: Process images, identify and count fluorescent spots, and calculate concentrations or convert them based on standard curves; S7. Result output: The concentration is displayed on the touch screen (4) and stored, exported or transmitted through the computing communication module (30); S8. Inspection Completion and Reset: Reset after inspection and clean the support platform (14).

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