CN114821578A - Microsphere counting method and system based on microscopic imaging and electronic equipment - Google Patents
Microsphere counting method and system based on microscopic imaging and electronic equipment Download PDFInfo
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Abstract
The invention relates to a microsphere counting method, a microsphere counting system and electronic equipment based on microscopic imaging, wherein the method comprises the following steps: acquiring a first to-be-detected image of the to-be-detected microsphere solution in each to-be-counted area based on a microscopic imaging technology; preprocessing each first image to be detected to obtain a second image to be detected of each area to be counted; and counting each circular connected domain in each second image to be detected to obtain the number of each microsphere in the microsphere solution to be detected. According to the invention, the number of microspheres with different diameters is obtained by adopting a microscopic imaging technology, and the microscopic imaging technology is combined with a computer vision technology, so that the defects of low resolution, narrow application range and the like of the existing microsphere counting software are overcome, automatic identification, classification and statistics of microsphere images are realized, the universality of a microscopic imaging microsphere counting platform is further widened, and the speed and the precision of analysis and detection are well improved.
Description
Technical Field
The invention relates to the technical field of image recognition, in particular to a microsphere counting method and system based on microscopic imaging and an electronic device.
Background
Most of the existing microsphere counting systems adopt biosensors for analysis and detection, such as electrochemical biosensors, mechanical biosensors, magnetic relaxation biosensors, and the like, and although the existing microsphere counting systems have the advantages of high sensitivity, high analysis speed, and the like, the signal reading mode and the signal reading equipment of the existing microsphere counting systems have many defects to be solved. The electrochemical biosensor usually needs to modify a working electrode, and the signal processing mode is complex, and a special signal acquisition card is needed; the mechanical biosensor has high requirements on the processing difficulty of the device and is difficult to produce in batch; the magnetic relaxation biosensor needs a special low-field nuclear magnetic resonance instrument which has strict requirements on temperature and large equipment volume, and is difficult to adapt to on-site rapid detection. The optical biosensor mainly uses the optical signal generated by the reaction of the substance to be detected and the detection reagent as the detection basis, and generally comprises 3 functional elements: the optical sensor comprises an optical sensing layer, an optical signal conversion element and a signal amplification processing element. However, the existing optical sensor has relatively high economic cost, precise structure, high signal processing requirement, and certain gap in stability compared with other biosensors.
Therefore, there is a need to develop a novel optical sensing platform, perfect biological functional substances, explore novel optical labeling materials or signal probes, and the like, which is significant for further development and application of optical biosensors.
Disclosure of Invention
In order to solve the technical problems, the invention provides a microsphere counting method and system based on microscopic imaging and an electronic device.
The invention discloses a microsphere counting method based on microscopic imaging, which comprises the following technical scheme:
s1, acquiring a first to-be-detected image of the to-be-detected microsphere solution in each to-be-counted area based on a microscopic imaging technology;
s2, preprocessing each first image to be detected to obtain a second image to be detected of each region to be counted, wherein each second image to be detected comprises at least one circular connected domain, and each circular connected domain corresponds to a microsphere with one diameter;
and S3, respectively counting each circular connected domain in each second image to be detected to obtain the number of each microsphere in the microsphere solution to be detected.
The microsphere counting method based on microscopic imaging has the following beneficial effects:
according to the method, the number of the microspheres with different diameters is obtained by adopting a microscopic imaging technology, and the microscopic imaging technology is combined with a computer vision technology, so that the defects of low resolution, narrow application range and the like of the existing microsphere counting software are overcome, automatic identification, classification and statistics of microsphere images are realized, the universality of a microscopic imaging microsphere counting platform is further widened, and the speed and the precision of analysis and detection are well improved.
On the basis of the scheme, the microsphere counting method based on microscopic imaging can be further improved as follows.
Further, the S2 specifically includes:
s21, performing gray-scale map conversion on each first image to be detected to obtain a first preprocessed image of each first image to be detected;
s22, performing binarization processing on each first pre-processed image to obtain a second pre-processed image of each first image to be detected;
and S23, filtering the interference information of each second preprocessed image to obtain each second image to be detected.
Further, the S23 specifically includes: setting a first threshold, and filtering interference information of each second preprocessed image according to the first threshold to obtain each second image to be detected; wherein the first threshold is predicted using a ResNet50 neural network model.
Further, the S3 specifically includes:
s31, marking each circular connected domain in each second image to be detected to obtain a marked image corresponding to each second image to be detected;
s32, identifying and counting the number of each marker in each marker image to obtain the number of each circular connected domain;
and S33, obtaining the number of each microsphere in the microsphere solution to be detected according to the number of each circular connected domain.
Further, each microsphere is no less than 2 microns in diameter.
The invention discloses a technical scheme of a microsphere technology system based on microscopic imaging, which comprises the following steps:
the method comprises the following steps: the device comprises a first processing module, a second processing module and a counting module;
the first processing module is configured to: acquiring a first to-be-detected image of the to-be-detected microsphere solution in each to-be-counted area based on a microscopic imaging technology;
the second processing module is configured to: preprocessing each first image to be detected to obtain a second image to be detected of each area to be counted, wherein each second image to be detected comprises at least one circular connected domain, and each circular connected domain corresponds to a microsphere with one diameter;
the counting module is used for: and respectively counting each circular connected domain in each second image to be detected to obtain the number of each microsphere in the microsphere solution to be detected.
The microsphere technology system based on microscopic imaging has the following beneficial effects:
the system of the invention obtains the number of microspheres with different diameters by adopting the microscopic imaging technology, and combines the microscopic imaging technology with the computer vision technology, thereby overcoming the defects of low resolution, narrow application range and the like of the existing microsphere counting software, realizing automatic identification, classification and statistics of microsphere images, further widening the universality of a microscopic imaging microsphere counting platform, and simultaneously well improving the speed and the precision of analysis and detection.
On the basis of the scheme, the microsphere technology system based on microscopic imaging can be further improved as follows.
Further, the second processing module specifically includes: the system comprises a first preprocessing module, a second preprocessing module and a third preprocessing module;
the first pre-processing module is configured to: performing gray-scale image conversion on each first image to be detected to obtain a first pre-processed image of each first image to be detected;
the second pre-processing module is configured to: carrying out binarization processing on each first pre-processed image to obtain a second pre-processed image of each first image to be detected;
the third pre-processing module is to: and filtering interference information of each second preprocessed image to obtain each second image to be detected.
Further, the counting module specifically includes: the device comprises a first counting module, a second counting module and a third counting module;
the first counting module is configured to: marking each circular connected domain in each second image to be detected to obtain a marked image corresponding to each second image to be detected;
the second counting module is configured to: identifying and counting the number of each type of mark in each mark image to obtain the number of each type of circular connected domain;
the third counting module is configured to: and obtaining the quantity of each microsphere in the microsphere solution to be detected according to the quantity of each circular connected domain.
The technical scheme of the storage medium of the invention is as follows:
the storage medium has stored therein instructions that, when read by a computer, cause the computer to perform the steps of a method for microsphere counting based on microscopic imaging according to the present invention.
The technical scheme of the electronic equipment is as follows:
comprising a memory, a processor and a computer program stored on the memory and executable on the processor, wherein the processor, when executing the computer program, causes the computer to perform the steps of a method for microsphere counting based on microscopic imaging according to the present invention.
Drawings
FIG. 1 is a schematic flow chart of a method for counting microspheres based on microscopic imaging according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of a microsphere counting system based on microscopic imaging according to an embodiment of the present invention.
Detailed Description
As shown in fig. 1, a first embodiment of a method for microsphere counting based on microscopic imaging according to the present invention comprises the steps of:
s1, acquiring a first image to be detected of the microsphere solution to be detected in each area to be counted based on the microscopic imaging technology.
Wherein, the microscopic imaging technology adopts the magnifying imaging function of a bright field optical microscope, and the model of the bright field optical microscope is as follows: LK-50, from the company Centipedae, Ltd. It should be noted that the imaging and capturing device may be a dark field optical microscope, a smart phone with a photographing function, a CMOS camera, and the like, besides the bright field optical microscope, and is not limited herein.
The microsphere solution in this embodiment is a polystyrene microsphere solution, and may also be any other polymeric microspheres, such as latex microspheres, silica microspheres, and the like, without limitation.
The polystyrene microsphere solution is prepared by using PS microspheres with diameters of 2 microns 2000 4 micron PS 4000 6 micron PS 6000 All carboxyl modified microspheres of (1) were purchased from Bangs Laboratories, Inc.
It should be noted that, the microspheres in this embodiment are carboxyl-modified microspheres with diameters of 2 micrometers, 4 micrometers, and 6 micrometers, and microspheres with diameters not smaller than 2 micrometers may also be used according to actual requirements, which is not limited herein.
Wherein, for the sake of explanation, PS is separately mixed with pure water 2000 Diluting to 1. mu.g/mL, 0.5. mu.g/mL, 0.25. mu.g/mL, 0.1. mu.g/mL, 0.05. mu.g/mL, 0.01. mu.g/mL in a gradient manner; PS Using pure Water 4000 Diluting to 10. mu.g/mL, 5. mu.g/mL, 2. mu.g/mL, 1. mu.g/mL, 0.5. mu.g/mL, 0.25. mu.g/mL in a gradient; using purePS with water 6000 Diluting to 30. mu.g/mL, 20. mu.g/mL, 10. mu.g/mL, 5. mu.g/mL, 1. mu.g/mL, 0.5. mu.g/mL in a gradient; the microsphere solution to be tested is according to PS 2000 ，PS 4000 ，PS 6000 At least one of the three is made of microspheres with any diameter and concentration. For example, PS is taken at a concentration of 0.5. mu.g/mL 2000 10mL, 5. mu.g/mL PS 4000 10mL, 20. mu.g/mL PS 6000 10ml of the microsphere solution to be detected is prepared.
Specifically, the microsphere solution to be detected is placed in an ultrasonic cleaner for 15 minutes of ultrasound. To ensure good observation, the coverslips were washed by immersion in an ethanol-water solution (75%, v/v) and wiped dry with a paper mirror. The test microsphere solution was dropped 10 μ L onto the counting area of a clean blood cell counting plate, and the wiped cover slip was gently covered, taking care not to generate bubbles during the process. Under the action of capillary phenomenon, the microsphere solution to be detected can fill the whole counting area. And (3) after standing for 3 minutes, naturally settling and stabilizing particles in the area to be counted so as to avoid counting errors caused by a layering phenomenon, placing the assembled slide under an optical microscope, selecting a 10-time ocular lens and a 20-time objective lens, and capturing images in the counting area on a computer screen through computer software (such as ImageView).
Wherein, the hemocytometer is purchased from Shanghai Biochemical reagent refining apparatus Co., Ltd; the model of the ultrasonic cleaner is BX5200HP, and is purchased from Shanghai New Miao medical instruments manufacturing Co., Ltd; ethanol was purchased from the national pharmaceutical group chemical agents limited.
In order to increase the accuracy of the method and the reliability of the result, the present embodiment selects to capture and identify the images of the microspheres in the two counting plates (four regions to be counted).
It should be noted that the number of counting regions can be set according to the requirement, and the more counting regions are captured, the higher the accuracy is, but the time cost is increased correspondingly.
Wherein the first image to be measured is an image of the microspheres within the area to be counted captured by computer software (ImageView).
S2, preprocessing each first image to be detected to obtain a second image to be detected of each region to be counted, wherein each second image to be detected comprises at least one circular connected domain, and each circular connected domain corresponds to a microsphere with one diameter.
Wherein the pre-treatment process substantially comprises: gray-scale map processing, binarization processing, image interference information filtering and the like.
Wherein, the second image to be measured is: comprises at least one circular connected domain, each circular connected domain has different diameter, and one circular connected domain corresponds to one diameter of the microsphere.
And S3, respectively counting each circular connected domain in each second image to be detected to obtain the number of each microsphere in the microsphere solution to be detected.
The microspheres are spherical, but are initially circular in a first image to be detected captured under an optical microscope, and can be more clearly distinguished from a second image to be detected, which is obtained by preprocessing the first image to be detected, so that the microspheres are circular connected domains in the second image to be detected.
And each circular connected domain adopts different marks, so that software can conveniently identify the number of each object to be identified.
After each circular connected domain is identified, the number of each circular connected domain is obtained, and classified statistics is carried out according to the number of each circular connected domain to obtain the number of each microsphere with each diameter; and finally, the data information in the image is classified and counted into a table for convenient reference.
Preferably, the S2 specifically includes:
and S21, performing grayscale map conversion on each first image to be detected to obtain a first preprocessed image of each first image to be detected.
The gray level image conversion of the first image to be detected has the following functions: the interference of natural light to the color information in the image is avoided.
Wherein the first pre-processed image is: and converting the gray level image of the first image to be detected to obtain an image.
And S22, performing binarization processing on each first pre-processed image to obtain a second pre-processed image of each first image to be detected.
The binarization processing of the first preprocessed image has the following functions: the gray value of the pixel point on the image is set to be 0 or 255, namely the whole image is subjected to an obvious black-and-white effect process, so that information in the first preprocessed image can be processed by a computer language conveniently.
Wherein the second preprocessed image is: and (3) obtaining an image after the first image to be detected is subjected to binarization processing.
And S23, filtering the interference information of each second preprocessed image to obtain each second image to be detected.
And each second image to be detected at least comprises a circular connected domain.
Preferably, the S23 specifically includes: setting a first threshold, and filtering interference information of each second preprocessed image according to the first threshold to obtain each second image to be detected; wherein the first threshold is predicted using a ResNet50 neural network model.
Specifically, each counting plate (counter) has a single background, and the target to be detected (circular connected domain) has an obvious difference in color depth from the background (the darker the color is, the smaller the gray value of the pixel is, the lighter the color is, and the larger the gray value of the pixel is), by using this characteristic, a first threshold is set, the pixel larger than the set threshold is set to be the maximum value 255, and the pixel smaller than the set threshold is set to be 0, so that noise point information can be filtered and the position information of the target to be detected can be focused on.
The set first threshold is generally set manually, and in the present embodiment, the neural network model ResNet50 is used for prediction, so as to replace the global threshold, thereby improving the detection accuracy.
Preferably, the S3 specifically includes:
and S31, marking each circular connected domain in each second image to be detected to obtain a marked image corresponding to each second image to be detected.
And S32, identifying and counting the number of each marker in each marker image to obtain the number of each circular connected domain.
S33, obtaining the number of each microsphere in the microsphere solution to be detected according to the number of each circular connected domain.
Preferably, each microsphere is no less than 2 microns in diameter.
Among these, microspheres are used as signal probes because they are observed when their size is 2 microns or larger.
In this embodiment, four to-be-counted areas are used by default for counting, classifying and identifying.
The microsphere counting method (system) of the embodiment has a sensitive and stable signal reading function, and can be applied to the fields of food safety rapid detection, in-vitro diagnosis, environmental monitoring and the like.
According to the technical scheme of the embodiment, the number of the microspheres with different diameters is obtained by adopting a microscopic imaging technology, and the microscopic imaging technology is combined with a computer vision technology, so that the defects of low resolution, narrow application range and the like of the existing microsphere counting software are overcome, automatic identification, classification and statistics of microsphere images are realized, the universality of a microscopic imaging microsphere counting platform is further widened, and the speed and the precision of analysis and detection are well improved.
In this example, by preparing microsphere solutions with a series of concentrations of 2 μm, 4 μm, 6 μm and a mixture of three diameters and counting the number of particles in the counting area through an optical microscope, it was found that when the concentration of the solution increases, the number of particles also increases, and a good correlation between the two is shown. By linear fit, the linear equation between the concentration of 2 μm, 4 μm, 6 μm microspheres and the corresponding number in the counting zone is: Y-450.86X-1.79 (R2-0.997), Y-39.822X +0.21 (R2-0.992), and Y-23.94X-5.38 (R2-0.993).
Meanwhile, counting the number of the microspheres in the counting area by adopting manual counting and QScounting, and the result shows that the manual counting has good consistency with the counting result of the microspheres based on microscopic imaging in the embodiment. Wherein, the microspheres with diameters of 4 μm and 6 μm are linearly fitted by taking the manual counting result as the abscissa and the counting result of the embodiment as the ordinate, and the linear equations are respectively as follows: y is 0.97X +1.11, Y is 0.97X +1.04, and the correlation coefficient (R2) reaches 0.999. For microspheres with a diameter of 2 μm, the linear equation is: y is 0.97X +0.16 and R2 is 0.996.
In summary, as the concentration of microspheres increases, the number of microspheres in the counting area increases. Through parameter optimization, the computer vision microsphere counting software also obtains good accuracy, and the analysis efficiency is well improved. Therefore, the microscopic imaging microsphere counting system manufactured by the method is a sensitive and stable signal reading device, and has great potential in the fields of food safety rapid detection, in-vitro diagnosis, environmental monitoring and the like.
The technical scheme of the embodiment has the advantages that: (1) the bright field optical microscope has mature manufacturing process, lower manufacturing cost and easy miniaturization; (2) different from fluorescent microspheres and the like, which need complex modification processes and strict storage conditions, the microspheres are used as a high polymer material, and have stable chemical properties and simple storage conditions; (3) microspheres with different diameters are combined for use, so that a new direction is provided for simultaneous detection of multiple targets; (4) the microscopic imaging platform is combined with a computer vision technology, so that automatic identification, classification and statistics of batch images can be realized, and the analysis and detection efficiency can be improved better.
In a second embodiment of the microsphere counting method based on microscopic imaging, based on the first embodiment, the technical solution of this embodiment further includes:
s100, acquiring a first image to be detected of the solution to be microsphericed in each area to be counted based on a microscopic imaging technology.
S200, preprocessing each first image to be detected to obtain a second image to be detected of each area to be counted, wherein each second image to be detected comprises at least one circular connected domain, and each circular connected domain corresponds to a microsphere with one diameter; wherein, the second image to be measured comprises: microspheres to be tested and free microspheres.
Wherein, the microsphere to be detected is: microspheres to be counted in this embodiment; the free microspheres are: no statistical microspheres are required in this example.
S300, counting each circular connected domain of the microspheres to be detected in each second image to be detected to obtain the number of each microspheres to be detected in the microsphere solution to be detected.
Specifically, for example, there exists a microsphere with a diameter of 6 microns in the second image to be measured, and several microspheres with a diameter of 2 microns are connected around the microsphere; the microspheres to be measured are: 2 micron microspheres attached to 6 micron microspheres, free microspheres are: 6 micron microsphere and 2 micron microsphere in free state; only the number of 2-micron microspheres attached to 6-micron microspheres in the image was counted using step S300.
It should be noted that, in this embodiment, the parameters such as the material and the diameter of the microsphere are not specifically required, and may be in any range.
The technical scheme of the embodiment can effectively reduce the steps of biochemical reaction and improve the speed of analysis and detection on the basis of the first embodiment.
As shown in fig. 2, a microsphere counting system 200 based on microscopic imaging according to an embodiment of the present invention includes: a first processing module 210, a second processing module 220, and a counting module 230;
the first processing module 210 is configured to: acquiring a first to-be-detected image of the to-be-detected microsphere solution in each to-be-counted area based on a microscopic imaging technology;
the second processing module 220 is configured to: preprocessing each first image to be detected to obtain a second image to be detected of each area to be counted, wherein each second image to be detected comprises at least one circular connected domain, and each circular connected domain corresponds to a microsphere with one diameter;
the counting module 230 is configured to: and respectively counting each circular connected domain in each second image to be detected to obtain the number of each microsphere in the microsphere solution to be detected, wherein one circular connected domain corresponds to a microsphere with one diameter.
Preferably, the second processing module 220 specifically includes: the system comprises a first pretreatment module, a second pretreatment module and a third pretreatment module;
the first pre-processing module is configured to: performing gray-scale map conversion on each first image to be detected to obtain a first preprocessed image of each first image to be detected;
the second pre-processing module is configured to: carrying out binarization processing on each first pre-processed image to obtain a second pre-processed image of each first image to be detected;
the third pre-processing module is to: and filtering interference information of each second preprocessed image to obtain each second image to be detected.
Preferably, the counting module 230 specifically includes: the device comprises a first counting module, a second counting module and a third counting module;
the first counting module is configured to: marking each circular connected domain in each second image to be detected to obtain a marked image corresponding to each second image to be detected;
the second counting module is configured to: identifying and counting the number of each type of mark in each mark image to obtain the number of each type of circular connected domain;
the third counting module is configured to: and obtaining the quantity of each microsphere in the microsphere solution to be detected according to the quantity of each circular connected domain.
According to the technical scheme of the embodiment, the number of the microspheres with different diameters is obtained by adopting a microscopic imaging technology, and the microscopic imaging technology is combined with a computer vision technology, so that the defects of low resolution, narrow application range and the like of the existing microsphere counting software are overcome, automatic identification, classification and statistics of microsphere images are realized, the universality of a microscopic imaging microsphere counting platform is further widened, and the speed and the precision of analysis and detection are well improved.
For the above steps for realizing the corresponding functions of each parameter and each module in the microsphere counting system 200 based on microscopic imaging according to this embodiment, reference may be made to each parameter and step in the first embodiment of the microsphere counting method based on microscopic imaging, which is not described herein again.
An embodiment of the present invention provides a storage medium, including: the storage medium stores instructions, and when the instructions are read by the computer, the computer is caused to execute the steps of the microsphere counting method based on microscopic imaging, which may specifically refer to the parameters and steps in the first embodiment of the microsphere counting method based on microscopic imaging, and are not described herein again.
Computer storage media such as: flash disks, portable hard disks, and the like.
An electronic device provided in an embodiment of the present invention includes a memory, a processor, and a computer program stored in the memory and executable on the processor, and is characterized in that when the processor executes the computer program, the computer executes a step of a microsphere counting method based on microscopic imaging, which may specifically refer to each parameter and step in the first embodiment of the microsphere counting method based on microscopic imaging, and is not described herein again.
Those skilled in the art will appreciate that the present invention may be embodied as methods, systems, storage media and electronic devices.
Thus, the present invention may be embodied in the form of: may be embodied entirely in hardware, entirely in software (including firmware, resident software, micro-code, etc.) or in a combination of hardware and software, and may be referred to herein generally as a "circuit," module "or" system. Furthermore, in some embodiments, the invention may also be embodied in the form of a computer program product in one or more computer-readable media having computer-readable program code embodied in the medium. Any combination of one or more computer-readable media may be employed. The computer readable medium may be a computer readable signal medium or a computer readable storage medium. A computer readable storage medium may be, for example, but not limited to, an electronic, magnetic, optical, electromagnetic, infrared, or semiconductor system, apparatus, or device, or any combination of the foregoing. More specific examples (a non-exhaustive list) of the computer readable storage medium would include the following: an electrical connection having one or more wires, a portable computer diskette, a hard disk, a Random Access Memory (RAM), a read-only memory (ROM), an erasable programmable read-only memory (EPROM or flash memory), an optical fiber, a portable compact disc read-only memory (CD-ROM), an optical storage device, a magnetic storage device, or any suitable combination of the foregoing. In the context of this document, a computer readable storage medium may be any tangible medium that can contain, or store a program for use by or in connection with an instruction execution system, apparatus, or device. Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (10)
1. A microsphere counting method based on microscopic imaging is characterized by comprising the following steps:
s1, acquiring a first to-be-detected image of the to-be-microsphere solution in each to-be-counted area based on a microscopic imaging technology;
s2, preprocessing each first image to be detected to obtain a second image to be detected of each region to be counted, wherein each second image to be detected comprises at least one circular connected domain, and each circular connected domain corresponds to a microsphere with one diameter;
s3, counting each circular connected domain in each second image to be detected to obtain the number of each microsphere in the microsphere solution to be detected.
2. The microsphere counting method based on microscopic imaging according to claim 1, wherein the S2 specifically comprises:
s21, performing gray-scale map conversion on each first image to be detected to obtain a first preprocessed image of each first image to be detected;
s22, performing binarization processing on each first pre-processed image to obtain a second pre-processed image of each first image to be detected;
and S23, filtering the interference information of each second preprocessed image to obtain each second image to be detected.
3. The microsphere counting method based on microscopic imaging according to claim 2, wherein the S23 specifically comprises: setting a first threshold, and filtering interference information of each second preprocessed image according to the first threshold to obtain each second image to be detected; wherein the first threshold is predicted using a ResNet50 neural network model.
4. The microsphere counting method based on microscopic imaging according to claim 1, wherein the S3 specifically comprises:
s31, marking each circular connected domain in each second image to be detected to obtain a marked image corresponding to each second image to be detected;
s32, identifying and counting the number of each marker in each marker image to obtain the number of each circular connected domain;
and S33, obtaining the number of each microsphere in the microsphere solution to be detected according to the number of each circular connected domain.
5. The microscopic imaging based counting method of claim 4, wherein each microsphere has a diameter of no less than 2 microns.
6. A microsphere counting system based on microscopic imaging, comprising: the device comprises a first processing module, a second processing module and a counting module;
the first processing module is configured to: acquiring a first to-be-detected image of the to-be-detected microsphere solution in each to-be-counted area based on a microscopic imaging technology;
the second processing module is configured to: preprocessing each first image to be detected to obtain a second image to be detected of each area to be counted, wherein each second image to be detected comprises at least one circular connected domain, and each circular connected domain corresponds to a microsphere with one diameter;
the counting module is used for: and counting each circular connected domain in each second image to be detected to obtain the number of each microsphere in the microsphere solution to be detected.
7. The microscopic imaging based microsphere counting system according to claim 6, wherein said second processing module comprises: the system comprises a first preprocessing module, a second preprocessing module and a third preprocessing module;
the first pre-processing module is to: performing gray-scale map conversion on each first image to be detected to obtain a first preprocessed image of each first image to be detected;
the second pre-processing module is configured to: carrying out binarization processing on each first pre-processed image to obtain a second pre-processed image of each first image to be detected;
the third pre-processing module is to: and filtering interference information of each second preprocessed image to obtain each second image to be detected.
8. The microscopic imaging based microsphere counting system according to claim 6, wherein said counting module comprises: the device comprises a first counting module, a second counting module and a third counting module;
the first counting module is configured to: marking each circular connected domain in each second image to be detected to obtain a marked image corresponding to each second image to be detected;
the second counting module is configured to: identifying and counting the number of each type of mark in each mark image to obtain the number of each type of circular connected domain;
the third counting module is configured to: and obtaining the quantity of each microsphere in the microsphere solution to be detected according to the quantity of each circular connected domain.
9. A storage medium having stored therein instructions which, when read by a computer, cause the computer to perform the microscopic imaging based microsphere counting method according to any one of claims 1 to 5.
10. An electronic device comprising a memory, a processor and a computer program stored on the memory and executable on the processor, wherein the processor, when executing the computer program, causes the computer to perform the method of microscopic imaging based counting of microspheres according to any of the claims 1 to 5.
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