Microsphere counting plate calibration method, microsphere counting plate, microsphere counting plate calibration device, cell counter calibration method, cell counter and computer readable storage medium

Abstract

The invention relates to the field of biomedical treatment, and discloses a microsphere counting plate calibration method, a microsphere counting plate calibration device, a cell counter calibration method, a cell counter and a computer readable storage medium. The microsphere counting plate calibration method comprises the following steps: determining a plurality of reference microsphere images from counting plate images; calculating according to the image position parameters to obtain a microsphere position relationship of the plurality of reference microsphere images; positioning a reference point in the counting panel image according to the image position parameter; constructing a counting area based on the reference point, and determining the number of microspheres in the counting area; and calibrating the microsphere counting plate by using the position relationship of the microspheres and the number of the microspheres. Compared with the prior art, the microsphere counting plate calibration method, the microsphere counting plate calibration device, the cell counter calibration method, the cell counter and the computer readable storage medium provided by the embodiment of the invention have the advantages that the calibration cost of the cell counter can be reduced, and the accuracy of a calibration result is improved.

Description

Technical Field

[0001] The present application relates to the field of biomedical technology, and more particularly, to a method for calibrating a microsphere counting plate, a microsphere counting plate, a device for calibrating a microsphere counting plate, a method for calibrating a cell counter, a cell counter, and a computer-readable storage medium. Background Art

[0002] In various clinical and research processes, cell counting is of interest, including the counting of white blood cells and red blood cells, which is valuable in the diagnosis of various diseases or abnormal conditions and in monitoring patients undergoing treatment for such diseases or conditions. Cells can be manually counted through the following process: placing a known sample dilution between optically clear plates that are sufficiently close to each other (usually, the spacing is on the order of 100 micrometers) so that the cells form a single layer; magnifying a region of the layer by a known multiple in a specified dimension; and counting the cells in the magnified region through a microscope. Due to the small size of cells, manual cell counters not only have low accuracy and are prone to counting errors when performing cell counting, but also cause discomfort to the user's eyes after long-term observation of the small-sized cells. On this basis, the cell counter was born. A cell counter is an instrument that can automatically detect the number of sample cells and is widely used in cell biology research in major universities and hospitals. However, as a metering device, the cell counter needs to be calibrated regularly to ensure the precision of the measurement.

[0003] When calibrating a cell counter, the prior art usually uses customized consumables with special cell-like markings made on them as a standard counting plate. This requires special customization, and the customized consumables have a limited number of uses, resulting in a high calibration cost. Moreover, since the standard counting plate for calibration is usually randomly placed, there is usually a certain range of variation in the standard counting plate within the field of view, resulting in the possibility that some regions of the standard counting plate are outside the field of view. The markings on these regions outside the field of view will not be counted, affecting the accuracy of the cell counter during calibration. Summary of the Invention

[0004] The purpose of the present application is to provide a method for calibrating a microsphere counting plate, a device for calibrating a microsphere counting plate, a method for calibrating a cell counter, a cell counter, and a computer-readable storage medium, which can reduce the calibration cost of the cell counter while improving the accuracy of the calibration result.

[0005] In a first aspect, an embodiment of the present application provides a method for calibrating a microsphere counting plate, including: obtaining a counting plate image of the microsphere counting plate; determining a plurality of reference microsphere images from the counting plate image, and respectively obtaining image parameters of each of the reference microsphere images, where the image parameters at least include image position parameters; calculating the microsphere position relationship of the plurality of reference microsphere images according to the image position parameters; positioning reference points in the counting plate image according to the image position parameters; constructing a counting area based on the reference points, and determining the number of microspheres in the counting area; and calibrating the microsphere counting plate using the microsphere position relationship and the number of microspheres.

[0006] Compared with the related art, in the method for calibrating a microsphere counting plate provided by the embodiment of the present application, after constructing a counting area in each microsphere counting plate and counting the number of microspheres in the counting area, the microsphere counting plate is calibrated using the microsphere position relationship of the counting area and the counted number of microspheres. Subsequently, when the cell counter calibrates using this microsphere counting plate, it can determine the counting area according to the calibrated microsphere position relationship, and compare the number of microspheres measured in the counting area with the calibrated number of microspheres to complete the calibration process of the cell counter. Since the calibrated microsphere counting plate can be reused, the calibration cost of the cell counter can be reduced; since the counting area is delimited using the microsphere position relationship of some reference microspheres in the counting plate image, that is, the counting area only includes a partial area in the counting plate image, even if there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position during the calibration process of the cell counter, resulting in an incomplete counting plate image taken during the calibration process, the counting area probably still exists in the taken counting plate image, and the calibration of the cell counter can still be accurately completed, reducing the impact of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and improving the accuracy of the calibration result of the cell counter.

[0007] In an optional embodiment, the obtaining a counting plate image of the microsphere counting plate includes: respectively obtaining counting plate images when the microsphere counting plate is in a plurality of different positions; the determining a plurality of reference microsphere images from the counting plate image includes: respectively determining the plurality of reference microsphere images from each of the counting plate images. Determining a plurality of reference microsphere images from the counting plate images of the microsphere counting plate in a plurality of different positions, that is, the determined plurality of reference microsphere images exist in all the counting plate images, that is, the counting plate images taken when the microsphere counting plate is in different positions all include reference microsphere images, which can increase the possibility of capturing all the reference microsphere images when there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position, reduce the impact of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and further improve the accuracy of the calibration result of the cell counter.

[0008] In an alternative embodiment, constructing the counting area based on the reference points includes: constructing the counting area in each of the counting plate images; if the counting area constructed in any of the counting plate images is incomplete, determining new reference microsphere images from each of the counting plate images until the counting areas constructed in all the counting plate images are complete. Redetermining new reference microsphere images when the counting area constructed in any of the counting plate images is incomplete can ensure that all the counting plate images include complete counting areas, that is, the counting plate images taken at different positions of the microsphere counting plate all include complete counting areas, which can increase the possibility of capturing a complete counting area when there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position, reduce the influence of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and further improve the accuracy of the calibration result of the cell counter.

[0009] In an alternative embodiment, determining the number of microspheres in the counting area includes: determining the number of microspheres in the counting area in each of the counting plate images; if the number of microspheres corresponding to each of the counting plate images is not completely the same, determining new reference microsphere images from each of the counting plate images until the number of microspheres corresponding to all the counting plate images is completely the same. When the number of microspheres corresponding to each of the counting plate images is not completely the same, there may be microspheres in the counting area that are prone to misjudgment. Redetermining new reference microsphere images until the number of microspheres corresponding to all the counting plate images is completely the same can ensure the accuracy of the calibrated number of microspheres. The higher the accuracy of the calibrated number of microspheres, the higher the accuracy of the calibration result when calibrating the cell counter.

[0010] In an alternative embodiment, calculating the microsphere position relationship of the plurality of reference microsphere images according to the image position parameters includes: determining any one of the reference microsphere images as the target microsphere image; calculating the position differences between each of the reference microsphere images and the target microsphere image. In the microsphere counting plate, the possibility that the position differences between multiple microspheres are the same is relatively low. Calculating the position differences between each of the reference microsphere images and the target microsphere image as the microsphere position relationship can more accurately locate the reference microsphere images. When calibrating the cell counter, the higher the positioning accuracy of the reference microsphere images, the higher the accuracy of the calibration result.

[0011] In an alternative embodiment, the image parameters further include an image size parameter and / or an image brightness parameter, and the method for calibrating the microsphere counting plate further includes: establishing a correspondence between the position difference and the image size parameter and / or the image brightness parameter; and calibrating the microsphere counting plate using the microsphere position relationship and the number of microspheres, including: calibrating the microsphere counting plate using the position difference, the image size parameter and / or the image brightness parameter, the correspondence, and the number of microspheres. In a microsphere counting plate, the possibility of several microspheres having the same position difference, the same image size parameter, and / or the same image brightness parameter is relatively low. Using the position difference, the image size parameter and / or the image brightness parameter, and the correspondence to calibrate the microsphere counting plate can more accurately locate the reference microsphere image. When calibrating a cell counter, the higher the positioning accuracy of the reference microsphere image, the higher the accuracy of the calibration result.

[0012] In an alternative embodiment, determining several reference microsphere images from the counting plate image includes: determining several microsphere image sets from the counting plate image, each microsphere image set including several reference microsphere images; calculating the microsphere position relationship of the several reference microsphere images according to the image position parameters includes: for each microsphere image set, calculating the microsphere position relationship corresponding to the microsphere image set according to the image position parameters of each reference microsphere image in the microsphere image set; locating a reference point in the counting plate image according to the image position parameters includes: for each microsphere image set, locating the reference point corresponding to the microsphere image set in the counting plate image according to the image position parameters of each reference microsphere image in the microsphere image set; and constructing a counting area based on the reference point includes: constructing the counting area based on the reference points corresponding to all the microsphere image sets.

[0013] In a second aspect, an embodiment of the present application provides a microsphere counting plate, including: a substrate and several microspheres fixed in the substrate.

[0014] In a third aspect, an embodiment of the present application provides a method for calibrating a cell counter, including: obtaining a calibration counting plate image and calibration information of a microsphere counting plate, where the calibration information at least includes the microsphere position relationship and the number of microspheres; obtaining calibration image parameters of each microsphere image in the calibration counting plate image, where the calibration image parameters at least include calibration position parameters; determining a plurality of calibrated microsphere images that meet the microsphere position relationship in the calibration counting plate image based on the calibration position parameters; positioning a reference point in the calibration counting plate image according to the calibration position parameters of the plurality of calibrated microsphere images; constructing a counting area based on the reference point, counting the microspheres in the counting area to obtain a calibration number; and determining the calibration result of the cell counter according to the calibration number and the number of microspheres.

[0015] Compared with the related art, in the method for calibrating a cell counter provided by the embodiment of the present application, by determining a corresponding counting area in the captured calibration counting plate image according to the microsphere position relationship in the calibration information of the microsphere counting plate, and performing microsphere counting on the microspheres in the counting area to obtain a calibration number, and then comparing the actual calibration number with the number of microspheres in the calibration information to determine the calibration result of the cell counter. Since the counting area only includes a part of the calibration counting plate image, even if there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position during the calibration process of the cell counter, resulting in an incomplete calibration counting plate image captured during the calibration process, the counting area is still likely to exist in the captured calibration counting plate image, and the calibration of the cell counter can still be accurately completed, reducing the influence of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and improving the accuracy of the calibration result of the cell counter.

[0016] In a fourth aspect, an embodiment of the present application provides a microsphere counting plate calibration device, including: an image acquisition module for acquiring a counting plate image of a microsphere counting plate; an image processing module for determining a plurality of reference microsphere images from the counting plate image, and respectively acquiring image parameters of each of the reference microsphere images, where the image parameters at least include image position parameters; the image processing module is further configured to calculate the microsphere position relationship of the plurality of reference microsphere images according to the image position parameters; the image processing module is further configured to position a reference point in the counting plate image according to the image position parameters; a counting module for constructing a counting area based on the reference point and determining the number of microspheres in the counting area; and a calibration module for calibrating the microsphere counting plate using the microsphere position relationship and the number of microspheres.

[0017] Fifth aspect, an embodiment of the present application provides a cell counter, including: an image acquisition module configured to acquire a calibration counting board image of a microsphere counting board; a calibration acquisition module configured to acquire calibration information of the microsphere counting board, where the calibration information includes at least the microsphere position relationship and the number of microspheres; a positioning module configured to acquire calibration image parameters of each microsphere image in the calibration counting board image, where the calibration image parameters include at least calibration position parameters; the positioning module is further configured to determine a plurality of calibrated microsphere images that satisfy the microsphere position relationship in the calibration counting board image based on the calibration position parameters; the positioning module is further configured to locate a reference point in the calibration counting board image according to the calibration position parameters of the plurality of calibrated microsphere images; a counting module configured to construct a counting area based on the reference point, count the microspheres in the counting area, and obtain a calibration quantity; a calibration module configured to determine a calibration result of the cell counter according to the calibration quantity and the number of microspheres.

[0018] Sixth aspect, an embodiment of the present application provides a computer-readable storage medium storing a computer program, where the computer program is executed by a processor to implement the microsphere counting board calibration method or the cell counter calibration method as described above. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] To more clearly illustrate the technical solutions of the embodiments of the present application, the following briefly introduces the drawings required to be used in the embodiments. It should be understood that the following drawings only show some embodiments of the present application and should not be regarded as limiting the scope. For those of ordinary skill in the art, other related drawings can also be obtained based on these drawings.

[0020] Figure 1 It is a schematic flowchart of the microsphere counting board calibration method provided by Embodiment 1 of the present application;

[0021] Figure 2 It is a schematic diagram of a counting board picture in the microsphere counting board calibration method provided by Embodiment 1 of the present application;

[0022] Figure 3 It is a schematic diagram of a counting board picture in the microsphere counting board calibration method provided by another embodiment of the present application;

[0023] Figure 4 、 Figure 5 is Figure 2 a partial enlarged view of;

[0024] Figure 6 It is a schematic structural diagram of the microsphere counting board calibration device provided by Embodiment 3 of the present application;

[0025] Figure 7 Schematic flowchart of the cell counter calibration method provided in the fourth embodiment of the present application;

[0026] Figure 8 Schematic structural diagram of the cell counter provided in the fifth embodiment of the present application. Detailed implementation manners

[0027] To make the objectives, technical solutions, and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described in detail below with reference to the accompanying drawings in the embodiments of the present application. Apparently, the described embodiments are some but not all of the embodiments of the present application. Usually, the components of the embodiments of the present application described and illustrated herein can be arranged and designed in various different configurations.

[0028] Therefore, the following detailed description of the embodiments of the present application provided in the drawings is not intended to limit the scope of the claimed present application, but merely represents selected embodiments of the present application.

[0029] It should be noted that similar reference numerals and letters denote similar items in the following drawings. Therefore, once an item is defined in one drawing, it does not need to be further defined and explained in subsequent drawings.

[0030] In addition, terms such as "first" and "second" are used for descriptive purposes only and cannot be construed as indicating or implying relative importance.

[0031] It should be noted that, without conflict, the features in the embodiments of the present application can be combined with each other.

[0032] Embodiment 1

[0033] Embodiment 1 of the present application provides a calibration method for a microsphere counting plate, which is used to calibrate the microsphere counting plates provided in other embodiments of the present application. The calibrated microsphere counting plate can be used to calibrate the counting accuracy of a cell counter. As Figure 1 shown, the calibration method for the microsphere counting plate provided in Embodiment 1 includes:

[0034] Step S101: Obtain a counting plate image of the microsphere counting plate.

[0035] Step S102: Determine a plurality of reference microsphere images from the counting plate image, and respectively obtain the image parameters of each reference microsphere image. The image parameters at least include image position parameters.

[0036] Step S103: Calculate the microsphere position relationships of the plurality of reference microsphere images according to the image position parameters.

[0037] Step S104: Locate the reference point in the counting plate image according to the image position parameters.

[0038] Step S105: Construct a counting area based on the reference point and determine the number of microspheres within the counting area.

[0039] Step S106: Calibrate the microsphere counting plate using the microsphere position relationship and the number of microspheres.

[0040] Among them, in the first embodiment, the counting plate image of the microsphere counting plate obtained in step S101 can specifically be the counting plate image obtained by placing the microsphere counting plate on a cell counter used for calibrating the microsphere counting plate and using the cell counter for calibration to photograph the microsphere counting plate. Or in some other embodiments of the present application, it can also be the counting plate image obtained by photographing the entire microsphere counting plate through other means, or it can be the counting plate image obtained in advance by communicating with communication devices such as servers through communication, etc., and other means to obtain the counting plate image of the microsphere counting plate.

[0041] Further, as Figure 2 shown, the counting plate image obtained in step S101 includes the microsphere image 100 of the microspheres in the microsphere counting plate. In different embodiments of the present application, for different types of microsphere counting plates, there may also be differences between the microsphere images 100 in the counting plate image. For example Figure 2 shown in the counting plate image, the sizes of different microsphere images 100 may be different (such as Figure 2 there are microsphere images 101 and 102 with different sizes), and the microsphere images 100 of different sizes correspond to microspheres of different sizes in the microsphere counting plate. The microspheres of different sizes in the microsphere counting plate can be used to simulate cells of different sizes, thereby expanding the application range of the microsphere counting plate.

[0042] It can be understood that the difference in the sizes between different microsphere images 100 in the counting plate image is only an example of the microsphere image 100 in some embodiments of the present application. In some other embodiments of the present application, as Figure 3 shown, the shapes of different microsphere images 100 in the counting plate image (such as Figure 3 there are a hollow microsphere image 103 and a solid microsphere image 104) can also be different. Microsphere images of the same shape can also be used to simulate cells of the same type and different states to expand the application range of the microsphere counting plate.

[0043] In step S102, the reference microsphere image is the microsphere image used to locate the counting area. In the first embodiment, as Figure 2 、 Figure 4 、 Figure 5As shown, determining several reference microsphere images from the counting plate image can specifically be determining several microsphere image sets from the counting plate picture, and each microsphere image set includes several reference microsphere images. For example Figure 2 , Figure 4 , Figure 5 As shown, three reference microsphere images a1, a2, and a3 together form a microsphere image set 1, and three reference microsphere images b1, b2, and b3 together form a microsphere image set 2.

[0044] Furthermore, in the first embodiment, obtaining the microsphere image set can specifically be obtaining several microsphere images that meet the preset conditions as reference microsphere images to jointly form a microsphere image set. Among them, the preset conditions can specifically include conditions such as the number, size, distance, and relative position of the reference microsphere images. For example Figure 2 , Figure 4 As shown, in the first embodiment, the preset conditions can be, for example, that the number of microsphere images meets three, the sizes of two of the three microsphere images are the same, the size of the other microsphere image is larger than the two microsphere images with the same size, the minimum distance between the three microsphere images is less than the preset distance, and the three microsphere images are arranged in a straight line. For another example Figure 2 , Figure 5 As shown, in the first embodiment, the preset conditions can also be, for example, that the number of microsphere images meets three, the sizes of the three microsphere images are the same, the three microsphere images are in contact, and the three microsphere images are arranged in a triangle.

[0045] It can be understood that the foregoing determination of several microsphere images that meet the preset conditions to jointly form a microsphere image set is only an example of the method for determining several reference microsphere images from the counting plate image in the first embodiment. In some other embodiments of the present application, for example, a human-computer interaction device can also be set up to determine several reference microsphere images from the counting plate image by interacting with the user, that is, determining several microsphere images specified by the user from the counting plate image to jointly form a microsphere image set.

[0046] Furthermore, in step S102, the image position parameter is specifically the position parameter of each microsphere image in the coordinate system constructed for the counting plate image. Among them, the coordinate system can, for example, construct a rectangular coordinate system with the center point of the counting plate image as the origin, and then determine the coordinate position of each reference microsphere image in the coordinate system as the image position parameter of each reference microsphere image.

[0047] In addition, in some embodiments of the present application, in addition to the image position parameter, the image parameters can also include, for example, the image size parameter, image shape parameter, image color parameter, image brightness parameter, etc. of the microsphere image as described above, and can be flexibly utilized according to the form of the microspheres in the microsphere counting plate.

[0048] In step S103, the specific microsphere position relationships of a number of reference microsphere images calculated based on the image position parameters can be as follows: For each set of microsphere images, the microsphere position relationship corresponding to the set of microsphere images is calculated according to the image position parameters of each reference microsphere image in the set of microsphere images. That is, each set of microsphere images corresponds to a microsphere position relationship.

[0049] Further, for each set of microsphere images, the specific calculation of the corresponding microsphere position relationship may include: determining any reference microsphere image as the target microsphere image; and respectively calculating the position differences between each reference microsphere image and the target microsphere image. As Figure 4 shown, for the three reference microsphere images a1, a2, and a3, for example, the reference microsphere image a1 can be determined as the target microsphere image, and the position differences between the three reference microsphere images a1, a2, and a3 and the target microsphere image are respectively calculated. In the microsphere counting plate, the possibility that the position differences between multiple microspheres are the same is relatively low. Calculating the position differences between each reference microsphere image and the target microsphere image as the microsphere position relationship can more accurately locate the reference microsphere images. When calibrating the cell counter, the higher the positioning accuracy of the reference microsphere images, the higher the accuracy of the calibration result.

[0050] Further, when the aforementioned image parameters may also include parameters such as the image size parameter, image shape parameter, image color parameter, and image brightness parameter of the aforementioned microsphere images, a corresponding relationship between the position difference and the image size parameter and / or image brightness parameter can also be constructed. On this basis, representing the three reference microsphere images a1, a2, and a3 in the form of an array is: ; wherein, the first column of the array is the abscissa of the reference microsphere image in the coordinate system, the second column is the ordinate of the reference microsphere image in the coordinate system, the third column is the image size parameter of the reference microsphere image, and the fourth column is the image shape parameter of the reference microsphere image (h represents a solid microsphere image). On this basis, calculating the position differences between each reference microsphere image and the target microsphere image is: .

[0053] As Figure 5 shown, for the three reference microsphere images b1, b2, and b3, for example, the reference microsphere image b1 can be determined as the target microsphere image, and the position differences between the three reference microsphere images b1, b2, and b3 and the target microsphere image are respectively calculated.

[0054] Further, when the aforementioned image parameters further include parameters such as the image size parameter, image shape parameter, image color parameter, and image brightness parameter of the aforementioned microsphere image, a corresponding relationship between the position difference and the image size parameter and / or image brightness parameter can also be constructed. On this basis, representing the three reference microsphere images b1, b2, and b3 in the form of an array is as follows: ; On this basis, calculating the position difference between each reference microsphere image and the target microsphere image is as follows: .

[0057] In the microsphere counting plate, the possibility of several microspheres having the corresponding relationship between the same position difference, the same image size parameter, and / or the same image brightness parameter is relatively low. Using the position difference, the image size parameter, and / or the image brightness parameter, and the corresponding relationship to calibrate the microsphere counting plate can more accurately locate the reference microsphere image. When calibrating the cell counter, the higher the positioning accuracy of the reference microsphere image, the higher the accuracy of the calibration result.

[0058] In step S104, specifically, locating the reference point in the counting plate image according to the image position parameter can be: for each microsphere image set, locating the reference point corresponding to the microsphere image set in the counting plate picture according to the image position parameters of each reference microsphere image in the microsphere image set. That is, each microsphere image combination corresponds to a reference point.

[0059] Further, in the first embodiment of the present application, specifically determining the reference point corresponding to each microsphere image set can be determining the center point of the target microsphere image in the microsphere image set as the reference point. Or in some other embodiments of the present application, it can also be determining the center point of all the reference microsphere images in the microsphere image set (the point corresponding to the average value of the position parameters of all the reference microsphere images) as the reference point, etc.

[0060] In step S105, specifically constructing the counting area based on the reference point can be constructing the counting area based on the reference points corresponding to all the microsphere image sets. As Figure 2 shown, in the first embodiment of the present application, specifically, a rectangular counting area is constructed with the two reference points corresponding to the two microsphere image sets as the relative vertices. Or in some other embodiments of the present application, it can also be constructing a circular counting area with the line connecting the two reference points corresponding to the two microsphere image sets as the diameter, or when including three, four... reference points, connecting the respective reference points in sequence to form corresponding triangular, quadrilateral... counting areas, etc.

[0061] In step S106, when the image parameters only include the image position parameters, the microsphere counting plate can be calibrated directly using the microsphere position relationship and the number of microspheres. When the image parameters further include other parameters such as the image size parameter, image shape parameter, image color parameter, and image brightness parameter of the microsphere image, the microsphere counting plate can also be calibrated using the position difference, image size parameter and / or image brightness parameter, corresponding relationship, and the number of microspheres.

[0062] It can be understood that in the first embodiment, the number of counting plate images can be one or more. When the number of counting plate images is more than one, obtaining the counting plate pictures of the microsphere counting plate in step S101 can be: obtaining the counting plate pictures when the microsphere counting plate is at several different positions respectively. Among them, different positions correspond to different positions of the microsphere counting plate on the cell counter used for calibration. Specifically, the microsphere counting plate can be placed at different positions on the cell counter used for calibration, and the cell counter used for calibration takes pictures to obtain several counting plate pictures when the microsphere counting plate is at several different positions.

[0063] Correspondingly, when the number of counting plate images is more than one, in step S102, determining several reference microsphere images from the counting plate pictures can specifically be: determining several reference microsphere images from each of the counting plate pictures respectively. That is, obtaining the microsphere images existing in each counting plate picture as the reference microsphere images. Determining several reference microsphere images from the counting plate pictures of the microsphere counting plate at several different positions, that is, the determined several reference microsphere images exist in all the counting plate pictures. That is, the counting plate pictures taken when the microsphere counting plate is at different positions all include the reference microsphere images, which can increase the possibility of capturing all the reference microsphere images when there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position, reduce the influence of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and further improve the accuracy of the calibration result of the cell counter.

[0064] Further, in some embodiments of the present application, when the number of counting plate images is multiple, the step of constructing a counting area based on the reference points in step S105 may further include: constructing a counting area in each counting plate image respectively; if the counting area constructed in any one of the counting plate images is incomplete, determining new reference microsphere images from each of the counting plate images until the counting areas constructed in all the counting plate images are complete. That is, for each counting plate image, a counting area is constructed in each counting plate image respectively, and then it is judged whether there is any counting area constructed in any one of the counting plate images that is incomplete, that is, the counting area cannot be completely constructed in this counting plate image. If there is any counting area constructed in any one of the counting plate images that is incomplete, new reference microsphere images are determined again from each of the counting plate images, and a new counting area is constructed again until the counting areas constructed in all the counting plate images are complete, that is, there is no situation where the counting area cannot be completely constructed in any counting plate image. When the counting area constructed in any one of the counting plate images is incomplete, determining new reference microsphere images again can ensure that all the counting plate images include complete counting areas, that is, the counting plate images taken at different positions of the microsphere counting plate all include complete counting areas, which can increase the possibility of capturing a complete counting area when there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position, reduce the influence of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and further improve the accuracy of the calibration result of the cell counter.

[0065] Further, in some embodiments of the present application, when the number of counting plate images is multiple, the step of determining the number of microspheres in the counting area in step S105 may further include: determining the number of microspheres in the counting area in each counting plate image respectively; if the number of microspheres corresponding to each counting plate image is not completely the same, determining new reference microsphere images from each of the counting plate images until the number of microspheres corresponding to all the counting plate images is completely the same. That is, the number of microspheres in the counting area of each counting plate image is counted respectively to obtain the number of microspheres corresponding to each counting plate image, and then all the numbers of microspheres corresponding to all the counting plate images are compared to judge whether all the numbers of microspheres are completely the same. If there is at least one counting plate image whose corresponding number of microspheres is different from that of other counting plate images, new reference microsphere images are determined again from each of the counting plate images, and a new counting area is constructed again, and the new number of microspheres is counted until the number of microspheres corresponding to all the counting plate images is completely the same. When the number of microspheres corresponding to each counting plate image is not completely the same, there may be microspheres in the counting area that are prone to misjudgment. Determining new reference microsphere images again until the number of microspheres corresponding to all the counting plate images is completely the same can ensure the accuracy of the calibrated number of microspheres. The higher the accuracy of the calibrated number of microspheres, the higher the accuracy of the calibration result when calibrating the cell counter.

[0066] Compared with the related art, in the microsphere counting plate calibration method provided by the first embodiment, after constructing a counting area in each microsphere counting plate and counting the number of microspheres in the counting area, the microsphere counting plate is calibrated using the microsphere position relationship in the counting area and the counted number of microspheres. Subsequently, when the cell counter calibrates using this microsphere counting plate, it can determine the counting area according to the calibrated microsphere position relationship, and compare the number of microspheres measured in the counting area with the calibrated number of microspheres to complete the calibration process of the cell counter. Since the calibrated microsphere counting plate can be reused, the calibration cost of the cell counter can be reduced; since the counting area is delimited using the microsphere position relationship of some reference microspheres in the counting plate image, that is, the counting area only includes a partial area in the counting plate image, even if there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position during the calibration process of the cell counter, resulting in an incomplete counting plate image taken during the calibration process, the counting area probably still exists in the taken counting plate image, and the calibration of the cell counter can still be accurately completed, reducing the impact of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result and improving the accuracy of the calibration result of the cell counter.

[0067] Embodiment 2

[0068] The second embodiment of the present application provides a microsphere counting plate, including: a substrate and a plurality of microspheres fixed in the substrate.

[0069] Among them, the plurality of microspheres may specifically include microspheres of different sizes and different colors. Among them, microspheres of different sizes can be used to simulate cells of different sizes (such as white blood cells and red blood cells), and microspheres of different colors can be used to simulate cells in different states (such as live cells and dead cells).

[0070] When preparing the microsphere counting plate provided by the second embodiment of the present application, it may specifically include:

[0071] Step 1: Determine the cell types and states to be simulated. The types are divided into small cell suspension, large cell suspension, and mixed suspension of small and large cells, and the states are divided into live cells, dead cells, and cells with different ratios of activity.

[0072] Step 2: Prepare a specific microsphere suspension according to the cell state to be simulated.

[0073] Step 3: Prepare a cell counting plate for use with a cell counter.

[0074] Step 4: Integrate and fix the microsphere suspension with the counting plate.

[0075] Compared with the related art, after being calibrated by the microsphere counting plate provided in the foregoing embodiments of the present application, the microsphere counting plate provided in the embodiments of the present application can be used to calibrate the counting accuracy of a cell counter, and can be reused during the calibration process, which better saves the cost of calibrating the cell counter.

[0076] Embodiment III

[0077] Embodiment III of the present application provides a method for calibrating a cell counter, as Figure 6 shown, including the following steps:

[0078] Step S201: Obtain a calibration counting plate image of the microsphere counting plate and calibration information, where the calibration information includes at least the microsphere position relationship and the number of microspheres.

[0079] Step S202: Obtain calibration image parameters of each microsphere image in the calibration counting plate image, where the calibration image parameters include at least calibration position parameters.

[0080] Step S203: Determine a plurality of calibrated microsphere images that satisfy the microsphere position relationship in the calibration counting plate image based on the calibration position parameters.

[0081] Step S204: Locate a reference point in the calibration counting plate picture according to the calibration position parameters of the plurality of calibrated microsphere images.

[0082] Step S205: Construct a counting area based on the reference point, count the microspheres in the counting area, and obtain a calibration quantity.

[0083] Step S206: Determine the calibration result of the cell counter according to the calibration quantity and the number of microspheres.

[0084] In this embodiment, the microsphere counting plate is the microsphere counting plate provided in the foregoing embodiments of the present application, and the calibration information is the data calibrated by the microsphere counting plate calibration method provided in the foregoing embodiments of the present application.

[0085] In step S206, the calibration quantity can be directly compared with the calibrated number of microspheres, and the calibration result of the cell counter is determined according to the difference between the calibration quantity and the calibrated number of microspheres. The smaller the difference between the two, the higher the counting accuracy of the cell counter.

[0086] It can be understood that in the cell counter calibration method provided in the third embodiment, steps S201 to S205 are substantially the same as steps S101 to S105 provided in the foregoing embodiments, and specific details can be referred to the specific descriptions in the foregoing embodiments. By adopting the same counting method as that in the calibration process of the microsphere counting plate provided in the foregoing embodiments during the calibration process of the cell counter, it can ensure that the acquisition process of the calibration quantity obtained during the calibration process of the cell counter is the same as that of the calibrated microsphere quantity. By counting the same microsphere counting plate in the same manner, the influence of the difference in counting methods on the counting results can be reduced, and the accuracy of the calibration results of the cell counter can be improved.

[0087] Compared with the related art, in the cell counter calibration method provided in the third embodiment of the present application, by determining the corresponding counting area in the captured calibration counting plate image according to the microsphere position relationship in the calibration information of the microsphere counting plate, and performing microsphere counting on the microspheres in the counting area to obtain the calibration quantity, and then comparing the actual calibration quantity with the microsphere quantity in the calibration information to determine the calibration result of the microsphere counter. Since the counting area only includes a part of the calibration counting plate image, even if there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position during the calibration process of the cell counter, resulting in an incomplete calibration counting plate image during the calibration process, the counting area is still likely to exist in the captured calibration counting plate image, and the calibration of the cell counter can still be accurately completed, reducing the influence of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and improving the accuracy of the calibration result of the cell counter.

[0088] Embodiment 4

[0089] The fourth embodiment of the present application provides a microsphere counting plate calibration device, as Figure 7 shown, including: an image acquisition module 701, which is used to acquire a counting plate picture of the microsphere counting plate; an image processing module 702, which is used to determine a plurality of reference microsphere images from the counting plate picture, and respectively acquire the image parameters of each reference microsphere image, and the image parameters at least include image position parameters; the image processing module 702 is further used to calculate the microsphere position relationship of a plurality of reference microsphere images according to the image position parameters; the image processing module 702 is further used to locate reference points in the counting plate picture according to the image position parameters; a counting module 703, which is used to construct a counting area based on the reference points and determine the number of microspheres in the counting area; a calibration module 704, which is used to calibrate the microsphere counting plate by using the microsphere position relationship and the number of microspheres.

[0090] Compared with the related art, in the microsphere counting plate calibration device provided in the fourth embodiment of the present application, after the image processing module 702 constructs a counting area in each microsphere counting plate and the counting module 703 counts the number of microspheres in the counting area, the calibration module 704 calibrates the microsphere counting plate using the microsphere position relationship in the counting area and the counted number of microspheres. Subsequently, when the cell counter uses this microsphere counting plate for calibration, it can determine the counting area according to the calibrated microsphere position relationship, and compare the number of microspheres measured in the counting area with the calibrated number of microspheres to complete the calibration process of the cell counter. Since the calibrated microsphere counting plate can be reused, the calibration cost of the cell counter can be reduced; since the counting area is delimited using the microsphere position relationship of some reference microspheres in the counting plate picture, that is, the counting area only includes a partial area in the counting plate picture, even if there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position during the calibration process of the cell counter, resulting in an incomplete counting plate picture taken during the calibration process, the counting area will still probably exist in the taken counting plate picture, and the calibration of the cell counter can still be accurately completed, reducing the impact of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and improving the accuracy of the calibration result of the cell counter.

[0091] Further, in some other embodiments of the present application, the image acquisition module 701 is further configured to respectively acquire counting plate pictures when the microsphere counting plate is in several different positions; on this basis, the image processing module 702 is further configured to respectively determine several reference microsphere images from each counting plate picture.

[0092] Further, in some other embodiments of the present application, when the image acquisition module 701 is configured to respectively acquire counting plate pictures when the microsphere counting plate is in several different positions, the image processing module 702 is further configured to respectively construct a counting area in each counting plate picture; if the counting area constructed in any one of the counting plate pictures is incomplete, the image processing module 702 is further configured to determine new reference microsphere images from each counting plate picture until the counting areas constructed in all the counting plate pictures are complete.

[0093] Further, in some other embodiments of the present application, when the image acquisition module 701 is configured to respectively acquire counting plate pictures when the microsphere counting plate is in several different positions, the counting module 703 is further configured to respectively determine the number of microspheres in the counting area of each counting plate picture; if the number of microspheres corresponding to each counting plate picture is not completely the same, the image processing module 702 is further configured to determine new reference microsphere images from each counting plate picture until the number of microspheres corresponding to all the counting plate pictures is completely the same.

[0094] Embodiment Five

[0095] Embodiment 5 of the present application provides a cell counter, as Figure 8 shown, including: an image acquisition module 801, which is used to acquire a calibration counting plate image of a microsphere counting plate; a calibration acquisition module 802, which is used to acquire calibration information of the microsphere counting plate, and the calibration information at least includes the microsphere position relationship and the number of microspheres; a positioning module 803, which is used to acquire calibration image parameters of each microsphere image in the calibration counting plate image, and the calibration image parameters at least include calibration position parameters; the positioning module 803 is further used to determine a plurality of calibrated microsphere images that meet the microsphere position relationship in the calibration counting plate image based on the calibration position parameters; the positioning module 803 is further used to locate a reference point in the calibration counting plate image according to the calibration position parameters of the plurality of calibrated microsphere images; a counting module 804, which is used to construct a counting area based on the reference point, count the microspheres in the counting area, and obtain a calibration quantity; a calibration module 805, which is used to determine the calibration result of the microsphere counter according to the calibration quantity and the number of microspheres.

[0096] Compared with the related art, in the cell counter provided in Embodiment 5 of the present application, the positioning module 803 determines a corresponding counting area in the captured calibration counting plate image according to the microsphere position relationship in the calibration information of the microsphere counting plate, and the counting module 804 counts the microspheres in the counting area to obtain a calibration quantity. Then, the calibration module 805 compares the actual calibration quantity with the number of microspheres in the calibration information to determine the calibration result of the microsphere counter. Since the counting area only includes a part of the calibration counting plate image, even if there is a deviation between the actual placement position of the microsphere counting plate and the standard calibration position during the calibration process of the cell counter, resulting in an incomplete calibration counting plate image captured during the calibration process, the counting area probably still exists in the captured calibration counting plate image, and the calibration of the cell counter can still be accurately completed, reducing the influence of the deviation between the actual placement position of the microsphere counting plate and the standard calibration position on the calibration result, and improving the accuracy of the calibration result of the cell counter.

[0097] Embodiment 6

[0098] Embodiment 6 of the present application relates to a computer-readable storage medium storing a computer program. When the computer program is executed by a processor, the above method embodiments are implemented.

[0099] That is, those skilled in the art can understand that all or part of the steps in the methods of the above embodiments can be completed by instructing relevant hardware through a program. The program is stored in a storage medium, including several instructions to enable a device (which can be a single-chip microcomputer, a chip, etc.) or a processor to execute all or part of the steps in the methods of the various embodiments of the present application. The aforementioned storage medium includes: various media such as USB flash drives, mobile hard disks, read-only memories (ROMs), random access memories (RAMs), magnetic disks, or optical discs that can store program codes.

[0100] The above is only the specific implementation manner of the present application, but the protection scope of the present application is not limited thereto. Any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed in the present application should be covered by the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A microsphere counting plate calibration method, characterized in that: include: Acquire a counting plate image of a microsphere counting plate; Determine a plurality of reference microsphere images from the counting plate image, and respectively obtain image parameters of each of the reference microsphere images, wherein the image parameters at least include image position parameters; Calculating the microsphere position relationship of the plurality of reference microsphere images according to the image position parameters; Locating a reference point in the counting plate image according to the image position parameter; Constructing a counting area based on the reference point, and determining the number of microspheres in the counting area; The microsphere counting plate is calibrated using the microsphere position relationship and the microsphere quantity.

2. The microsphere counting plate calibration method according to claim 1, characterized in that: The step of obtaining a counting plate image of a microsphere counting plate comprises: Respectively acquiring images of the microsphere counting plate when the microsphere counting plate is in several different positions; Determining a plurality of reference microsphere images from the counting plate image comprises: The plurality of reference microsphere images are determined respectively from each of the counting plate images.

3. The microsphere counting plate calibration method according to claim 2, characterized in that: The constructing a counting area based on the reference point comprises: Constructing the counting area in each of the counting plate images respectively; If the counting area constructed in any of the counting plate images is incomplete, a new reference microsphere image is determined from each of the counting plate images until the counting areas constructed in all of the counting plate images are complete.

4. The microsphere counting plate calibration method according to claim 3, characterized in that: Determining the number of microspheres in the counting area comprises: respectively determining the number of microspheres in the counting area in each of the counting plate images; If the numbers of microspheres corresponding to the respective counting plate images are not exactly the same, a new reference microsphere image is determined from the respective counting plate images until the numbers of microspheres corresponding to all the counting plate images are exactly the same.

5. The microsphere counting plate calibration method according to claim 1, characterized in that: The step of calculating the microsphere position relationship of the plurality of reference microsphere images according to the image position parameters includes: Determining any one of the reference microsphere images as a target microsphere image; The position differences between each of the reference microsphere images and the target microsphere image are calculated respectively.

6. The microsphere counting plate calibration method according to claim 5, characterized in that: The image parameters also include image size parameters and / or image brightness parameters, and the microsphere counting plate calibration method also includes: Constructing a corresponding relationship between the position difference and the image size parameter and / or the image brightness parameter; The method of calibrating the microsphere counting plate using the microsphere position relationship and the microsphere quantity comprises: The microsphere counting plate is calibrated using the position difference, the image size parameter and / or the image brightness parameter, the corresponding relationship and the number of microspheres.

7. The microsphere counting plate calibration method according to claim 1, characterized in that: Determining a plurality of reference microsphere images from the counting plate image comprises: Determining a plurality of microsphere image sets from the counting plate image, each of the microsphere image sets including a plurality of reference microsphere images; The step of calculating the microsphere position relationship of the plurality of reference microsphere images according to the image position parameters includes: For each of the microsphere image sets, a microsphere position relationship corresponding to the microsphere image set is calculated according to image position parameters of each of the reference microsphere images in the microsphere image set; The step of locating a reference point in the counting plate image according to the image position parameter comprises: For each of the microsphere image sets, locating a reference point corresponding to the microsphere image set in the counting plate image according to image position parameters of each of the reference microsphere images in the microsphere image set; The constructing a counting area based on the reference point comprises: The counting area is constructed based on the reference points corresponding to all the microsphere image sets.

8. A microsphere counting plate, characterized in that: include: A substrate and a plurality of microspheres fixed in the substrate.

9. A cell counter calibration method, characterized in that: include: Acquire a calibration counting plate image and calibration information of a microsphere counting plate, wherein the calibration information at least includes a microsphere position relationship and a microsphere quantity; Acquire calibration image parameters of each microsphere image in the calibration counting plate image, wherein the calibration image parameters at least include calibration position parameters; Determining a plurality of calibration microsphere images satisfying the microsphere position relationship in the calibration counting plate image based on the calibration position parameters; Locating reference points in the calibration counting plate image according to calibration position parameters of the plurality of calibration microsphere images; Constructing a counting area based on the reference point, counting the microspheres in the counting area to obtain a calibration number; A calibration result of the cell counter is determined based on the calibration quantity and the microsphere quantity.

10. A microsphere counting plate calibration device, characterized in that: include: An image acquisition module, wherein the image acquisition module is used to acquire a counting plate image of a microsphere counting plate; An image processing module, the image processing module is used to determine a plurality of reference microsphere images from the counting plate image, and respectively obtain image parameters of each of the reference microsphere images, wherein the image parameters at least include image position parameters; The image processing module is also used to calculate the microsphere position relationship of the plurality of reference microsphere images according to the image position parameters; The image processing module is also used to locate the reference point in the counting plate image according to the image position parameter; A counting module, the counting module is used to construct a counting area based on the reference point and determine the number of microspheres in the counting area; A calibration module is used to calibrate the microsphere counting plate using the microsphere position relationship and the microsphere quantity.

11. A cell counter, characterized in that: include: An image acquisition module, wherein the image acquisition module is used to acquire a calibration counting plate image of a microsphere counting plate; A calibration acquisition module, the calibration acquisition module is used to acquire calibration information of the microsphere counting plate, the calibration information at least includes the microsphere position relationship and the microsphere quantity; A positioning module, the positioning module is used to obtain calibration image parameters of each microsphere image in the calibration counting plate image, the calibration image parameters at least including calibration position parameters; The positioning module is also used to determine a plurality of calibration microsphere images satisfying the microsphere position relationship in the calibration counting plate image based on the calibration position parameters; The positioning module is also used to locate the reference point in the calibration counting plate image according to the calibration position parameters of the plurality of calibration microsphere images; A counting module, the counting module is used to construct a counting area based on the reference point, count the microspheres in the counting area, and obtain a calibration number; A calibration module is used to determine a calibration result of the cell counter according to the calibration quantity and the microsphere quantity.

12. A computer-readable storage medium storing a computer program, characterized in that: The computer program is executed by a processor to implement the microsphere counting plate calibration method according to any one of claims 1 to 8 or the cell counter calibration method according to claim 9.

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