Fluorescence calibration method and device and computer readable storage medium

Abstract

The invention relates to the technical field of medical instruments, and provides a fluorescence calibration method and device.The fluorescence calibration method is applied to in-vitro diagnostic equipment, and the in-vitro diagnostic equipment comprises a carrier used for loading a detected object and a photographing module used for photographing the carrier; the fluorescence calibration method comprises the following steps: acquiring a first detection image of a carrier and a detected object shot by a shooting module based on a white light source; acquiring a second detection image of the carrier and the detected object shot by the shooting module based on the fluorescent light source; confirming a carrier area of the carrier and a read value of the detected object in the carrier area based on the first detection image and the second detection image; a corrected detection value of the read value in the carrier region is determined on the basis of a calibration curve of the read value in the carrier region. According to the invention, the container with uneven light receiving can obtain the reading value under the condition of uniform light receiving, the deviation coefficient is reduced, and the precision of the interpretation result is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and in particular to a fluorescence calibration method and device. Background Art

[0002] In in vitro diagnostic equipment, fluorescence interpretation is one of the commonly used interpretation methods. After the excitation light of a specific wavelength generated by the optical system is irradiated on the reaction cup, the magnetic beads and enhancers in the reaction cup are irradiated by the excitation light of the specific wavelength. The luminescent substances adsorbed on the magnetic beads absorb the light energy and enter the excited state, and then emit fluorescence longer than the laser wavelength. The external camera takes a picture of the reaction cup area to obtain a fluorescence image.

[0003] For the fluorescence images obtained in the prior art, the light source hardware such as lamp beads is generally adjusted to achieve the purpose of uniform distribution of pixel values ​​in the fluorescence image. However, in actual application, refer to Figure 2 The light formed by the excitation light irradiating the reaction cup is radial, and the pixel values ​​of the generated fluorescence image are not evenly distributed. This phenomenon is due to the difference in the light received by the reaction cup caused by the installation of the optical system itself, and the differences that may be caused by the aging of the optical system components. For example, the brightness of the lamp changes with the length of use, which leads to a large accuracy error between the interpreted fluorescence image and the actual image. Summary of the invention

[0004] In order to overcome the deficiencies of the prior art, the object of the present invention is to provide a fluorescence standard method and device to solve the problems of low accuracy and large coefficient of deviation of the interpretation results of the interpretation methods in the prior art.

[0005] In a first aspect, the present invention provides a fluorescence calibration method, which is applied to an in vitro diagnostic device, wherein the in vitro diagnostic device includes a carrier for loading an object to be detected and a camera module for photographing the carrier, and the fluorescence calibration method includes: Based on a white light source, obtaining a first detection image of the carrier and the detected object taken by the camera module; Based on the fluorescent light source, obtaining a second detection image of the carrier and the detected object taken by the camera module; Based on the first detection image and the second detection image, confirming the carrier area of ​​the carrier and the reading value of the detected object in the carrier area; According to a calibration curve of the read value in the carrier area, a corrected detection value of the read value in the carrier area is determined.

[0006] The fluorescence calibration method provided by the embodiment of the first aspect of the present invention has at least the following beneficial effects: the present invention utilizes the correspondence between the pixel value of the acquired fluorescence image and the actual brightness of the fluorescent substance in the actual image in the calibration curve to interpret each pixel point in the fluorescence image and obtain an effective correction detection value actually related to the fluorescent object, so that the detection and interpretation information of the actual distribution state of the fluorescent object in the fluorescence image is not affected by the uneven light exposure of the reaction cup, the deviation coefficient is reduced, the accuracy of the interpretation result is increased, and it can be applied to the phenomenon of reaction cup light exposure deviation caused by reasons such as the position deviation of the optical system of the interpretation module of different equipment.

[0007] In a preferred embodiment of the present invention, the process of obtaining the calibration curve includes: Acquire a calibration image, where the calibration image is an image taken by the camera module on a carrier loaded with a mixture of a magnetic substance and a fluorescent substance; Confirming the carrier range in the calibration image and determining the reference point with the highest reading value in the carrier range; the carrier range is the same as the carrier area; According to the position of the reference point in the carrier area, a calibration curve of the read value at the corresponding position in the carrier area is determined.

[0008] In a preferred embodiment of the present invention, the calibration image includes a first image and a second image, and acquiring the calibration image includes: Based on a white light source, obtaining a first image of the carrier and the mixture taken by the camera module; Based on the fluorescent light source, obtaining a second image of the carrier and the mixture taken by the camera module; The confirming the carrier range in the calibration image comprises: Confirming area information of the carrier in the first image according to brightness information of the carrier in the first image; The fluorescent area in the second image is determined according to the brightness information of the fluorescent substance in the second image, and the carrier range of the carrier in the second image is confirmed in combination with the area information of the carrier in the first image.

[0009] In a preferred embodiment of the present invention, the determining of the reference point with the highest reading value in the carrier range comprises: Dividing the carrier range into a plurality of pixel points, wherein each of the pixel points corresponds to a first pixel value capable of reflecting area information; Selecting an area consisting of a set number of pixel points having the highest first pixel value as a reference area; The highest pixel point is selected from the reference area as the reference point.

[0010] In a preferred embodiment of the present invention, determining the calibration curve of the read value at the corresponding position in the carrier region according to the position of the reference point in the carrier range includes: Determining a reference line within the carrier based on the reference point; Determine a calibration pixel value of any pixel point on the reference line according to the pixel value of each pixel point in the carrier range and the positional relationship with the pixel points on the reference line; A calibration curve is obtained according to the calibration pixel value of each pixel point on the reference line and the positional relationship between each pixel point and the reference point.

[0011] In a preferred embodiment of the present invention, the step of determining a reference line within the carrier range based on the reference point comprises: The reference line is a straight line connecting a reference point and a relative point on the edge of the carrier range, wherein the relative point is a point on the edge of the carrier range that is farthest from the reference point; Determining the calibration pixel value of any pixel point on the baseline according to the pixel value of each pixel point in the carrier range and the positional relationship with the pixel points on the baseline includes: Take any pixel point on the baseline, and take the average of the first pixel values ​​of all the pixel points within the specified area with the pixel point as the center and located within the range of the carrier as the calibration pixel value of the pixel point; According to the calibration pixel value of each pixel point on the reference line and the positional relationship between each pixel point and the reference point, a calibration curve is obtained: A curve fitting is performed on the calibration pixel values ​​of all the pixel points on the reference line and the distance between the corresponding pixel point and the reference point to obtain a calibration curve.

[0012] In a preferred embodiment of the present invention, the set number of pixel points with the highest first pixel values ​​are selected in the following manner: Grouping the pixel points into a first set, and sorting the pixel points in the first set from large to small according to the first pixel value; Selecting a set number of pixel points that are located at the front when the first set is sorted as pixel points constituting the reference area; The selecting the highest pixel point from the reference area as the reference point comprises: Forming a pixel area with each pixel point in the reference area as the center, wherein one pixel area corresponds to a second pixel value; The pixel areas are sorted according to the second pixel values, starting from the pixel area corresponding to the largest second pixel value, and the central pixel points in the pixel areas are sequentially determined as reference points until the central pixel point is determined as the reference point.

[0013] In a preferred embodiment of the present invention, the forming of a pixel area with each pixel point in the reference area as the center comprises: Taking the same number of adjacent pixel points in multiple directions around the central pixel point, each of the adjacent pixel points and the central pixel point forms a pixel area; The second pixel value is obtained based on the first pixel value, and the obtaining method includes: The sum of the first pixel values ​​corresponding to all the pixel points in the pixel area is calculated, and the sum is selected as the second pixel value.

[0014] In a preferred embodiment of the present invention, the step of determining a reference point for a central pixel in the pixel area includes: In multiple directions around the pixel area to be judged, respectively judging whether the pixel values ​​of the same number of pixel points as the pixel points in the pixel area in each direction are less than the first pixel value corresponding to the central pixel point in the pixel area; If the first pixel values ​​corresponding to the central pixel point in the pixel area are all greater than the pixel values ​​of the pixels in all directions, the central pixel point corresponding to the pixel area is determined to be the reference point.

[0015] In a preferred embodiment of the present invention, the confirming the carrier area of ​​the carrier based on the first detection image and the second detection image includes: Confirming area information of the carrier in the first detection image according to brightness information of the carrier in the first detection image; Determine the fluorescent area in the second detection image according to the brightness information of the fluorescent substance in the second detection image, and confirm the carrier area of ​​the carrier in the second detection image in combination with the area information of the carrier in the first detection image; Confirming the read value of the detected object in the carrier area based on the first detection image and the second detection image includes: The carrier area is divided into a plurality of pixel points, wherein each pixel point corresponds to a pixel value that can reflect area information, and the read value is a second set of pixel values ​​of corresponding pixel points of the detected object in the second detection image.

[0016] In a preferred embodiment of the present invention, determining the corrected detection value of the read value in the carrier area according to a calibration curve of the read value in the carrier area includes: According to the distance between the pixel points corresponding to all the pixel values ​​of the second set and the reference point in the calibration curve, the corrected pixel value of each pixel point in the carrier area is obtained, and the corrected detection value is a set composed of each of the corrected pixel values.

[0017] In a second aspect, the present invention provides a fluorescence calibration device, which is applied to an in vitro diagnostic device, wherein the in vitro diagnostic device comprises a carrier for carrying an object to be detected and a camera module for photographing the carrier, and is characterized in that it comprises: An acquisition unit, used to acquire a calibration curve and a detection image of the carrier and the detected object taken by the camera module, wherein the detection image includes a first detection image acquired based on a white light source and a second detection image acquired based on a fluorescent light source; A reading unit, configured to confirm a carrier region of the carrier and a reading value of the detected object in the carrier region according to the detection module acquired by the acquisition unit; A calibration unit is used to determine a corrected detection value of the read value in the carrier area according to a calibration curve of the read value in the carrier area.

[0018] In a third aspect, the present invention further provides a computer-readable storage medium, wherein the computer-readable storage medium stores a program, and when the program runs, the device controls the computer-readable storage medium to execute the method provided in the embodiment of the first aspect.

[0019] Other features and advantages of the invention will be described in the following description, and partly become apparent from the description, or understood by implementing the technical solution of the invention. The purpose and other advantages of the invention can be achieved and obtained by the structures and / or processes specifically pointed out in the description, claims and drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 A schematic diagram of the structure of an in vitro diagnostic device provided by an embodiment of the present invention;

[0021] Figure 2 A schematic diagram of light reception when the light reception inside the carrier provided by an embodiment of the present invention is uneven;

[0022] Figure 3 A flow chart of a fluorescence calibration method provided by an embodiment of the present invention;

[0023] Figure 4 A flowchart of a method for obtaining a calibration curve provided by an embodiment of the present invention;

[0024] Figure 5 It is a structural schematic diagram of a first image captured by a photographing module of an in vitro diagnostic device according to an embodiment of the present invention;

[0025] Figure 6 A schematic diagram of the structure of a second image captured by a camera module of an in vitro diagnostic device according to an embodiment of the present invention;

[0026] Figure 7 A schematic structural diagram of a fluorescence calibration device provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0027] The following will describe the implementation methods of the present invention in detail in combination with the accompanying drawings and embodiments, so that the implementation process of how the present invention applies technical means to solve technical problems and achieve technical effects can be fully understood and implemented accordingly. It should be noted that these specific descriptions are only for ordinary technicians in this field to understand the present invention more easily and clearly, rather than a restrictive interpretation of the present invention; for example, the first and second mentioned in the embodiments of the present invention do not constitute a limitation thereto, but are only for expressing the serial numbers of multiple identical or similar devices and mechanisms. Ordinary technicians in this field can also readjust these serial numbers for the convenience of expression or in the process of arranging technical solutions; and alternative solutions are described for some mechanisms in different embodiments, and these alternatives can also be applied to other identical or similar devices and mechanisms; and as long as there is no conflict, the various embodiments in the present invention and the various features in each embodiment can be combined with each other, and the technical solutions formed are all within the protection scope of the present invention.

[0028] The technical solution of the present invention is described in detail below through the accompanying drawings and specific embodiments.

[0029] First, the in vitro diagnostic device used in the embodiment of the present invention is described. Figure 1 The in vitro diagnostic device is used for a carrier 20 for loading the object to be detected and a camera module 10 for photographing the carrier 20. The object to be detected can reflect the light emitted by an external light source, or after being irradiated by a laser of a specific wavelength emitted by a fluorescent light source in an external optical path system, enter an excited state and emit fluorescent light with a wavelength longer than the specific wavelength of the laser to the outside, so as to be reflected by a reflector in the external optical path system. When the camera module 10 photographs the material in the carrier 20 in the reflector, a fluorescent image that can reflect the density and distribution position of the object to be detected is obtained. The carrier 20 is a storage container that can store the object to be detected, such as a reaction cup, a beaker or other container structure. The camera module 10 can use a camera in the prior art, or other photographing equipment, which can photograph the state in the carrier 20 and intercept photos during the shooting process to obtain the image required for interpretation. Of course, it can also be other photographing equipment that can obtain fluorescent images, which is not limited here.

[0030] In this embodiment, the object to be detected is magnetic beads, and the carrier 20 is set as a reaction cup. After the fluorescent substance in the reaction cup is adsorbed by the magnetic beads, the external excitation light excites the fluorescence of the fluorescent substance on the magnetic beads and is captured by the camera module 10, and is further interpreted and detected by the interpretation module in the in vitro diagnostic equipment.

[0031] Based on this, the present invention proposes a fluorescence calibration method in a first embodiment, which is applied to the above-mentioned in vitro diagnostic device, and can calibrate the interpretation image obtained by the camera module to obtain an effective detection value. Figure 3 , the fluorescence calibration method includes: S100, obtaining a first detection image of the carrier and the detected object captured by a camera module based on a white light source; S200, based on the fluorescent light source, obtaining a second detection image taken by the camera module of the carrier and the object to be detected.

[0032] Among them, the first light source is set as a white light source. Under the irradiation of the first light source, when the carrier is a container made of transparent materials such as a reaction cup, the camera module shoots the carrier and the object to be detected to obtain a first detection image. The background of the first detection image is in a white light state, and the area of ​​the carrier where the object to be detected is loaded will show a clear contour line in the first detection image. During the process of judging the first detection image, the judgment module can obtain the area information of the carrier in the first detection image based on the contour line, thereby obtaining the exact position of the carrier.

[0033] The second light source is set as a fluorescent light source, which emits a laser with a specific wavelength toward the area of ​​the carrier where the object to be detected is loaded, so that the object to be detected emits corresponding fluorescent light in the carrier. Then the camera module photographs the carrier and the object to be detected to obtain a second detection image. The background in the second detection image is dark. In this embodiment, the area information of the carrier cannot be obtained based on the second detection image, but it can correspond to the area information of the carrier obtained in the first detection image. The area information of the carrier will not change when only the first light source and the second light source are switched. Therefore, the first detection image is used to obtain the area information of the carrier, and the second detection image further obtains the detection information of the object to be detected based on the area information of the carrier during the interpretation process.

[0034] S300, based on the interpreted image, confirming the carrier area of ​​the carrier in the interpreted image and the reading value of the detected object in the carrier area; In the judgment image, the judgment image includes the first detection image and the second detection image, and it can be known that the carrier obtains the carrier area in the judgment image. As mentioned in the above description, the carrier area is obtained from the area information of the carrier area obtained in the first detection image, specifically including: S310, confirming area information of the carrier in the first detection image according to brightness information of the carrier in the first detection image; S320, determining the fluorescent area in the second detection image according to the brightness information of the fluorescent substance in the second detection image, and confirming the carrier area of ​​the carrier in the second detection image in combination with the area information of the carrier in the first detection image.

[0035] In the white field information corresponding to the first detection image, the carrier will appear as brightness information inconsistent with the background brightness in the first detection image. According to the brightness information, the contour line of the carrier can be confirmed, and the regional information corresponding to the carrier can be further divided based on the contour line. In one embodiment, when a carrier such as a reaction cup is placed in a white light source, the light irradiation direction of the white light source passes through the carrier from above the carrier, and then enters the shooting module to be photographed by the shooting module. At this time, since the light will have a certain amount of light loss after passing through the object to be detected and the bottom of the carrier, the area corresponding to the carrier will have a darker brightness image compared to other areas. According to the regional position of the pixel value corresponding to the image, the regional information of the carrier is obtained.

[0036] The second detection image corresponds to dark field information, and the carrier cannot be detected in the second detection image, but the fluorescence emitted by the detected object in the excited state can be captured by the camera module in the second detection image, and the fluorescence area is determined according to the brightness information corresponding to the captured fluorescence. In this embodiment, the position does not change during the light source switching process, thereby combining the area information of the carrier in the first detection image to further confirm the carrier area of ​​the carrier in the second detection image.

[0037] According to the above steps, the detected object in the judgment image can be read, including: The carrier area is divided into a plurality of pixel points, wherein each pixel point corresponds to a pixel value that can reflect the area information, and the read value is a second set of pixel values ​​of the corresponding pixel points of the detected object in the judgment image.

[0038] The carrier area can be considered to be composed of multiple pixels, and the pixel value on the pixel can correspond to the brightness information of the pixel in the second detection image. At this time, the read value is a second set of pixel values ​​of the corresponding pixel of the detected object in the read image. The second set is composed of multiple pixels with specific pixel values, forming a fluorescent image on the second detection image. The fluorescent image is the interpretation of the detected object with fluorescent substances in the second detection image. After the read value is calibrated, an accurate calibration detection value is obtained.

[0039] S400, determining a correction detection value of the read value in the carrier area according to a calibration curve of the read value in the carrier area.

[0040] The corrected detection value can be obtained based on the calibration curve. Specifically, after obtaining the read value of the corresponding fluorescent image in the corresponding carrier area, the correspondence between the position of any point of the corresponding fluorescent image on the calibration curve and the position of the corresponding point of the calibration curve in the carrier area is used to obtain the corrected detection value after correcting the read value. The corrected detection value is the actual detection value, which can reduce the reading deviation.

[0041] In one embodiment, the process of obtaining the correction detection value includes: According to the distance between the pixel points corresponding to all the pixel values ​​of the second set and the reference point in the calibration curve, the corrected pixel value of each pixel point in the carrier area is obtained, and the corrected detection value is a set composed of each corrected pixel value.

[0042] The calibration curve in this embodiment has a reference point, and there is a certain correspondence between the pixel value of the corresponding point at the corresponding distance from the reference point and the actual pixel value. It can correspond to a fluorescent image with a brightness radiation distribution, which has a center point and the farther away from the center point, the lower the brightness of the light source. Based on this, accurate correction pixel values ​​are obtained through the corresponding relationship with the reference point distance distribution, and the correction pixel values ​​are aggregated to form a correction detection value.

[0043] In addition, the calibration curve may also be a curve relationship corresponding to other non-uniform light sources, and the corresponding actual correction detection value may be obtained according to the curve relationship, which is within the scope of the implementation of the present invention.

[0044] Accordingly, in some embodiments, see Figure 4 , the process of obtaining the calibration curve may include: S500, acquiring a calibration image, where the calibration image is an image captured by a photographing module of a carrier loaded with a mixture of a magnetic substance and a fluorescent substance; In the process of acquiring the calibration image, a mixture containing a magnetic substance and a fluorescent substance is loaded into a carrier, and because the mixture can be in a liquid state in the carrier, it can be evenly mixed in the carrier accordingly, so that when acquiring the calibration image, especially in acquiring the fluorescent image obtained by the second light source, the carrier can determine the deviation state between the actual performance caused by the radial light distribution and the detection value in the fluorescent image by judging the pixel value of the fluorescent substance in the fluorescent image, and further obtain the corresponding brightness correction relationship based on the deviation state to form a calibration curve. Among them, the magnetic substance can absorb the fluorescent substance, and the fluorescent substance forms a larger area of ​​fluorescent area in the fluorescent image after being absorbed by the magnetic substance, so as to be further judged in the subsequent process.

[0045] In one embodiment, the mixture contains a small amount of magnetic substance and a large amount of fluorescent substance, wherein the magnetic substance is a magnetic bead without adsorbing the sample reagent, and the fluorescent substance is fluorescein. After each magnetic substance adsorbs more fluorescent substance, a larger light spot is formed in the fluorescent image. The uniform brightness state of the light spot is subsequently analyzed, and the reference point, i.e., the center point, in the radial light is obtained. It should be noted that the mixture and the object to be detected are composed of different substances. The fluorescent substance in the mixture can be set to a substance commonly used for detection and verification, such as fluorescein, which can be captured by the camera module and form a fluorescent image. The fluorescent substance in the object to be detected is the fluorescent substance represented by the corresponding component in the sample, and the fluorescent substance can also be adsorbed by the magnetic substance, so that the content of the corresponding component in the object to be detected can be known according to the corrected detection value in the second detection image taken by the camera module.

[0046] The calibration image includes a first image and a second image, the first image is obtained by a first light source as a white light source, and the second image is obtained by a second light source as a fluorescent light source, specifically including: S510, acquiring a first image of the carrier and the mixture captured by a camera module based on a white light source; S520, based on the fluorescent light source, obtaining a second image of the carrier and the mixture taken by the camera module.

[0047] The first image and the second image correspond to the first detection image and the second detection image. The first image is a white field image obtained based on a white light source. The reaction cup and other carriers have a relatively obvious outline in the white field image, and the outline corresponds to the carrier range of the carrier, so that the regional information of the carrier can be obtained in the first image. The second image is a dark field image obtained based on a fluorescent light source. The carrier range cannot be determined in the dark field image. Therefore, the fluorescent light source is switched while keeping the camera module and the carrier position unchanged. The acquired fluorescent information is based on the regional information of the carrier in the first image to know the corresponding position of the fluorescent substance of the corresponding point in the carrier range.

[0048] It should be noted that the first image and the second image are taken at the same viewing angle of the carrier, usually after being reflected by the reflector of the optical path system from the top view direction of the carrier, so as to better observe and judge the distribution of the fluorescent material in the carrier. Similarly, the first detection image and the second detection image are also arranged in this way, and no further explanation will be given here.

[0049] S600, confirming the carrier range in the calibration image, and determining the reference point with the highest reading value in the carrier range, the carrier range being the same as the carrier area; As shown above, after obtaining the regional information of the carrier in the first image, the carrier range of the carrier in the second image can be confirmed based on the regional information. The carrier range is the same as the carrier regions corresponding to the first detection image and the second detection image, thereby being able to confirm the corresponding positions of points including fluorescent points in the second image within the carrier range, so as to establish a calibration curve based on the information of each point at the position.

[0050] In one embodiment, the carrier range confirmation method includes: S610, confirming area information of the carrier in the first image according to brightness information of the carrier in the first image; S620, determining the fluorescent area in the second image according to the brightness information of the fluorescent substance in the second image, and confirming the carrier range of the carrier in the second image in combination with the area information of the carrier in the first image.

[0051] Similar to the first detection image, the carrier in the first image will also have its own structure and other reasons, causing the light transmitted by the white light source to be dissipated or completely reflected, so that the carrier has obvious contour information in the first image. The area information is further determined based on the contour information and the position in the first image. On this basis, the second image can be combined with the area information of the carrier in the first image to confirm the carrier range corresponding to the carrier. In addition, since the fluorescent substance appears as a light spot or point in the second image, when the second image assigns brightness information to each point on it, the position of the fluorescent substance in the second image can be known and the fluorescent information in the second image can be determined based on the position.

[0052] In this embodiment, when determining the reference point with the highest reading value in the carrier range, the following steps are specifically performed: S630, dividing the carrier range into a plurality of pixel points, wherein each pixel point corresponds to a first pixel value capable of reflecting the region information; S640, selecting an area consisting of a set number of pixel points with the highest first pixel value as a reference area; S650: Select the highest pixel point from the reference area as the reference point.

[0053] See also Figure 5, the carrier range represented by the carrier in the first image is set to D1, and accordingly, the carrier range is represented as image area D2 in the second image, and the range of the image area D2 is the same as the range of the carrier range D1. Subsequently, the carrier is subdivided into multiple small areas, each of which is regarded as a pixel point N1~Nn, each pixel point corresponds to a first pixel value that can reflect the regional information of the location of the pixel point, and the pixel point can be composed of one or more pixels in the second image, which is not limited here. In this embodiment, the number of pixels constituting the pixel point is the same and the shape is the same. The first pixel value of each pixel point can correspond to the average value or the sum of the pixel values ​​of the pixels constituting the pixel point, which is not limited here. In one embodiment, a pixel corresponds to a dot constituting the pixel point, and the pixel point is composed of multiple dots, wherein each dot is composed of adjacent dots in the front, back, left, and right directions to form a small set, and the small set constitutes a pixel point, and the first pixel value is the sum of the pixel values ​​corresponding to each dot, and then the reference point is analyzed and obtained based on the pixel point.

[0054] Each first pixel value corresponds to brightness information, and the brightness information may be obtained by graying the second image, or other parameters that can represent brightness information, all of which are within the scope of this embodiment. Thus, after assigning the brightness information as a numerical value to the first pixel value of each pixel point, a set number of pixel points with the highest light-receiving pixel value can be determined as reference points.

[0055] In the specific determination, in one implementation manner, the method of selecting a set number of pixel points with the highest first pixel value includes: S641, grouping the pixels into a first set, and sorting the pixels in the first set from large to small according to the first pixel value; S642, selecting a set number of pixel points that are located at the front in the first set during sorting as pixel points that constitute the reference area.

[0056] After the pixel points are organized into a first set, they are sorted from large to small according to the first pixel value to obtain a set number k of pixel points Nm1, Nm2, Nm3...Nmk with the largest first pixel value. Based on this, each pixel point in the first set, hereinafter referred to as the selected pixel point, is analyzed, and accurate reference points are obtained after excluding abnormal pixels from the selected pixel points.

[0057] In the specific analysis, a reference area D3 is formed based on each selected pixel point Nm1, Nm2, Nm3 ... Nmk, and after performing abnormality elimination analysis on the pixel points in each reference area D3, it is known whether the selected pixel point is a reference point.

[0058] The abnormality elimination analysis process specifically includes: S643, forming pixel areas with each pixel point in the reference area as the center, wherein one pixel area corresponds to one second pixel value; S644, sorting the pixel areas according to the second pixel values, starting from the pixel area corresponding to the largest second pixel value, sequentially determining the reference points of the central pixel points in the pixel areas until the central pixel point is determined as the reference point.

[0059] First, each pixel point in the reference area D3 is analyzed to determine whether the selected pixel point is an abnormal pixel point. In one embodiment, the selected pixel point is taken as the center, and the pixel points within a preset range around the selected pixel point as the center are taken to form a pixel area D4. Taking Nmk as an example, the same number of adjacent pixel points in multiple directions around the selected pixel point Nmk, such as up, down, front, back, front upper, front lower, back upper, and back lower, can be set to one or more, and can be reasonably set according to the specific carrier size and other specific conditions. In this embodiment, it is represented as n. Therefore, the pixel area D4 has 8n+1 pixel points with the selected pixel point Nmk as the center. For each pixel point in the 8n+1 pixel points, a pixel area is formed with each pixel point as the center, and the second pixel value is determined according to the first pixel value of each pixel point in the pixel area. The central pixel point in each pixel area is judged as a reference point starting from the largest second pixel value. When it meets the reference point requirements, it is judged as the central pixel point that can be used as the reference point in this embodiment.

[0060] According to the aforementioned embodiment, the method for forming the pixel region includes: S645, taking the same number of adjacent pixel points in multiple directions around the central pixel point, and each of the adjacent pixel points and the central pixel point form a pixel area.

[0061] The second pixel value is obtained from the first pixel value, and the obtaining method includes: The sum of the first pixel values ​​corresponding to all the pixel points in the pixel area is calculated, and the sum is selected as the second pixel value.

[0062] When the second pixel value is the sum of multiple first pixel values, the situation where the first pixel value of an individual pixel is abnormally high can be ruled out. That is, in the fluorescent image, a pixel that does not conform to the radiation direction may appear in the original radiation receiving scene due to shooting errors or other reasons. This abnormal point can be ruled out based on the sum of multiple pixel values.

[0063] One embodiment, when determining a reference point for a central pixel point in a pixel area, includes: S646, in multiple directions surrounding the pixel area to be determined, respectively determining whether the first pixel values ​​of the same number of pixels as the pixel points in the pixel area in each direction are smaller than the first pixel value corresponding to the central pixel point in the pixel area; S647: If the first pixel values ​​corresponding to the central pixel point in the pixel area are all greater than the pixel values ​​of the pixels in all directions, the central pixel point corresponding to the pixel area is determined to be the reference point.

[0064] Among them, continuing to use the above embodiment as a reference, when the number of pixels constituting the pixel area is 8n+1, the pixel values ​​of the 8n+1 pixel points in multiple directions surrounding the pixel area, such as up, down, front, back, front upper, front lower, back upper, and back lower, are judged in turn with the first pixel value of the central pixel point. If there is a pixel point whose pixel value is greater than or equal to the central pixel point, it means that the central pixel point is not the reference point with the highest pixel value, and the pixel area does not meet the requirements and enters the central pixel area of ​​the next pixel area with a lower second pixel value for reference point judgment. In other words, when the first pixel value of the central pixel point is greater than the pixel values ​​of all the pixel points used for judgment, it means that the pixel area and the adjacent pixel points are radially centered on the central pixel point, and thus the central pixel point can be judged as the reference point. It should be noted that the pixel value of the pixel point used for judgment in this embodiment is the first pixel value.

[0065] S700, determining a calibration curve of a reading value at a corresponding position in the carrier region according to the position of the reference point in the carrier region.

[0066] According to the above embodiment, after the reference point is determined, it is necessary to further determine the calibration curve of the reading value in the second image at the corresponding position in the carrier area. Because the reference point is determined, the fluorescence brightness of the radiant light centered on the reference point is in an inverse proportional relationship with the distance from the reference point, that is, the farther away from the reference point, the weaker the brightness that can be photographed by the camera module under the excited state of the same fluorescent substance. Therefore, the embodiment of the present invention uses the distance as the independent variable to obtain the calibration curve.

[0067] In one embodiment, after obtaining the position of the reference point, determining a calibration curve of the read value at the corresponding position in the carrier area includes: S710, determining a reference line within the carrier range based on the reference point; S720, determining a calibration pixel value of any pixel point on the reference line according to the pixel value of each pixel point in the carrier range and the positional relationship with the pixel points on the reference line; S730: Obtain a calibration curve according to the calibration pixel value of each pixel point on the reference line and the positional relationship between each pixel point and the reference point.

[0068] Within the carrier range, the reference line can correspond to the position of the pixel point corresponding to any position within the carrier range. With the reference line as a reference, the reference point is set at the starting position of the reference line and the reference line extends toward the edge of the carrier range until it intersects with the edge. Any pixel point on the reference line has a corresponding position distance with the reference point. Under the position distance, there is a corresponding relationship between the pixel value and the distance of the corresponding pixel point, and the calibration curve is determined according to the corresponding relationship.

[0069] The reference line can also be set as a line segment corresponding to the shortest distance between the reference point and the edge of the carrier range, or a line segment formed by connecting any of its extreme points within the carrier range. Because the affected area of ​​the light receiving range may be different, in special cases, there will be a light receiving deviation within a certain range within the carrier range, and this deviation may not exist after leaving the range. Therefore, the reference line can be specifically set according to the actual situation. In addition, in addition to being set as a straight line, the reference line can also be set as a curve. Because the difference in radiation-type light receiving may vary in distance, setting it to a relationship corresponding to its position distance can also achieve the above effect.

[0070] After determining the baseline, since the distance between any point within the range affected by the light deviation and the reference point can correspond to each other on the baseline, in the process of calibrating the read value by obtaining the second detection image in the aforementioned embodiment, the read value is corrected according to the positional correspondence between the pixel points on the second detection image and the reference point. Taking the baseline as a straight line as an example, when the radiation deviation under the influence of light deviation is a uniform deviation relationship, that is, the farther away from the reference point, the lower the pixel value of the captured image is, and the light deviation relationship between each point on the circular edge line at the same distance from the reference point and the reference point is determined, then in the second detection image, the corrected detection value of any point can be obtained through the positional relationship between each pixel point and the reference point.

[0071] In one embodiment, when the radiation type is affected by the light deviation within the entire carrier range, determining a reference line within the carrier range based on the reference point includes: S711, the reference line is a straight line connecting the reference point and the relative point on the edge of the carrier range, wherein the relative point is the point on the edge of the carrier range that is farthest from the reference point.

[0072] See also Figure 6 When the end point of the baseline is selected as the pixel point B farthest from the reference point A within the carrier range, the baseline can be used as the radiation range to cover all the pixels within the carrier range. Therefore, during the interpretation process, all the pixels on the second detection image can obtain the corrected detection value in the calibration curve according to the baseline.

[0073] In another embodiment, determining the calibration pixel value of any pixel point on the baseline according to the pixel value of each pixel point in the carrier range and the positional relationship with the pixel point on the baseline includes: S721, taking any pixel point on the baseline, and taking the average of the first pixel values ​​of all the pixels within the specified area centered on the pixel point and located within the carrier range as the calibration pixel value of the pixel point; S722, obtaining a calibration curve according to the calibration pixel value of each pixel point on the reference line and the positional relationship between each pixel point and the reference point: S723, performing curve fitting on the calibration pixel values ​​of all pixel points on the reference line and the distances between the corresponding pixel points and the reference points to obtain a calibration curve.

[0074] After obtaining the baseline, there may be abnormal pixels in any pixel on the baseline, so the pixel is excluded as an abnormality, and the average of the first pixel values ​​of all pixels in the carrier range within the designated area centered on the pixel is used as the baseline pixel value of the pixel. The designated area can be the circular area D5, which is centered on the pixel and has a radius of R, wherein the radius R may correspond to M pixels, that is, a circular area D5 composed of M pixels in all directions centered on the pixel, and the average of the pixel values ​​of each pixel in the circular area D5 is the baseline pixel value corresponding to the pixel on the baseline.

[0075] After obtaining the reference pixel value of any pixel point on the reference line one by one, the corresponding calibration curve can be obtained with distance as the independent variable according to the fitting relationship between the pixel value and the distance. In the subsequent interpretation process, the read value is corrected according to the distance relationship between the interpretation curve and the reference point.

[0076] In a second aspect, the present invention further provides a fluorescence calibration device 30, which is applied to an in vitro diagnostic device, wherein the in vitro diagnostic device includes a carrier 20 for loading an object to be detected and a camera module 10 for photographing the carrier, including: An acquisition unit 31 is used to acquire a calibration curve and a detection image taken by a camera module of the carrier and the detected object, wherein the detection image includes a first detection image acquired based on a white light source and a second detection image acquired based on a fluorescent light source; The reading unit 32 is used to confirm the carrier area of ​​the carrier and the reading value of the detected object in the carrier area according to the detection module obtained by the obtaining unit 31; The calibration unit 33 is used to determine the corrected detection value of the read value in the carrier area according to the calibration curve of the read value in the carrier area.

[0077] In a third aspect, an embodiment of the present invention further provides a computer-readable storage medium, the computer-readable storage medium comprising a stored program, wherein when the program is running, the computer-readable storage medium is controlled to execute the fluorescence calibration method of the embodiment of the first aspect of the present invention in a device.

[0078] The embodiment of the present invention further provides a computer device, the computer device of the embodiment comprises: a processor, a memory, and a computer program stored in the memory and executable on the processor, the computer program when executed by the processor implements the method in the embodiment, to avoid repetition, it is not described one by one here. Alternatively, the computer program when executed by the processor implements the functions of each model / unit of the control device in the embodiment, to avoid repetition, it is not described one by one here.

[0079] Computer devices include, but are not limited to, processors and memories. Those skilled in the art will appreciate that the above are merely examples of computer devices and do not constitute a limitation on computer devices. Computer devices may include more or fewer components than shown in the figure, or may combine certain components, or different components. For example, computer devices may also include input and output devices, network access devices, buses, etc.

[0080] The processor may be a central processing unit 33 (CPU), or other general-purpose processors, digital signal processors (DSP), application-specific integrated circuits (ASIC), field-programmable gate arrays (FPGA) or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc. A general-purpose processor may be a microprocessor or any conventional processor, etc.

[0081] The memory may be an internal storage unit of a computer device, such as a hard disk or memory of the computer device. The memory may also be an external storage device of the computer device, such as a plug-in hard disk, a smart media card (SMC), a secure digital (SD) card, a flash card, etc. equipped on the computer device. Furthermore, the memory may include both an internal storage unit of the computer device and an external storage device. The memory is used to store computer programs and other programs and data required by the computer device. The memory may also be used to temporarily store data that has been output or is to be output.

[0082] In the above embodiments, it can be implemented in whole or in part by software, hardware, firmware or any combination thereof. When implemented using software, it can be implemented in whole or in part in the form of a computer program product. The computer program product includes one or more computer instructions. When the computer instructions are loaded and executed on a computer, the process or function described in the embodiment of the present application is generated in whole or in part. The computer can be a general-purpose computer, a special-purpose computer, a computer network, or other programmable devices. The computer instructions can be stored in a computer-readable storage medium, or transmitted from one computer-readable storage medium to another computer-readable storage medium, for example, the computer instructions can be transmitted from a website site, computer, server or data center by wired (e.g., coaxial cable, optical fiber, digital subscriber line (Digital Subscriber Line, DSL)) or wireless (e.g., infrared, wireless, microwave, etc.) mode to another website site, computer, server or data center. The computer-readable storage medium can be any available medium that a computer can access or a data storage device such as a server, data center, etc. that contains one or more available media integrated. The available medium may be a magnetic medium (eg, a floppy disk, a hard disk, a magnetic tape), an optical medium (eg, a high-density digital video disc (DVD)), or a semiconductor medium (eg, a solid state disk (SSD)).

[0083] Although some specific embodiments of the present invention have been described in detail by way of example, it will be appreciated by those skilled in the art that the above examples are for illustration only and are not intended to limit the scope of the present invention. It will be appreciated by those skilled in the art that the above embodiments may be modified without departing from the scope and spirit of the present invention. The scope of the present invention is defined by the appended claims.

[0084] Finally, it should be noted that the above description is only the best embodiment of the present invention and does not limit the present invention in any form. Any technician familiar with the art can make many possible changes and simple replacements to the technical solution of the present invention by using the above disclosed practices and technical contents without departing from the scope of the technical solution of the present invention, which all belong to the scope of protection of the technical solution of the present invention.

Claims

1. A fluorescence calibration method, applied to an in vitro diagnostic device, wherein the in vitro diagnostic device comprises a carrier for carrying an object to be detected and a camera module for photographing the carrier, characterized in that: The fluorescence calibration method comprises: Based on a white light source, obtaining a first detection image of the carrier and the detected object taken by the camera module; Based on the fluorescent light source, obtaining a second detection image of the carrier and the detected object taken by the camera module; Based on the first detection image and the second detection image, confirming the carrier area of ​​the carrier and the reading value of the detected object in the carrier area; According to a calibration curve of the read value in the carrier area, a corrected detection value of the read value in the carrier area is determined.

2. The method according to claim 1, characterized in that The process of obtaining the calibration curve includes: Acquire a calibration image, where the calibration image is an image taken by the camera module on a carrier loaded with a mixture of a magnetic substance and a fluorescent substance; Confirming the carrier range in the calibration image and determining the reference point with the highest reading value in the carrier range; the carrier range is the same as the carrier area; According to the position of the reference point in the carrier area, a calibration curve of the read value at the corresponding position in the carrier area is determined.

3. The method according to claim 2, characterized in that The calibration image includes a first image and a second image, and acquiring the calibration image includes: Based on a white light source, obtaining a first image of the carrier and the mixture taken by the camera module; Based on the fluorescent light source, obtaining a second image of the carrier and the mixture taken by the camera module; The confirming the carrier range in the calibration image comprises: Confirming area information of the carrier in the first image according to brightness information of the carrier in the first image; The fluorescent area in the second image is determined according to the brightness information of the fluorescent substance in the second image, and the carrier range of the carrier in the second image is confirmed in combination with the area information of the carrier in the first image.

4. The method according to claim 2, characterized in that: The step of determining the reference point with the highest reading value in the carrier range includes: Dividing the carrier range into a plurality of pixel points, wherein each of the pixel points corresponds to a first pixel value capable of reflecting area information; Selecting an area consisting of a set number of pixel points having the highest first pixel value as a reference area; The highest pixel point is selected from the reference area as the reference point.

5. The method according to claim 4, characterized in that The step of determining a calibration curve of the read value at a corresponding position in the carrier region according to the position of the reference point in the carrier range comprises: Determining a reference line within the carrier based on the reference point; Determine a calibration pixel value of any pixel point on the reference line according to the pixel value of each pixel point in the carrier range and the positional relationship with the pixel points on the reference line; A calibration curve is obtained according to the calibration pixel value of each pixel point on the reference line and the positional relationship between each pixel point and the reference point.

6. The method according to claim 5, characterized in that The determining of a reference line within the carrier range based on the reference point comprises: The reference line is a straight line connecting a reference point and a relative point on the edge of the carrier range, wherein the relative point is a point on the edge of the carrier range that is farthest from the reference point; Determining the calibration pixel value of any pixel point on the baseline according to the pixel value of each pixel point in the carrier range and the positional relationship with the pixel points on the baseline includes: Take any pixel point on the baseline, and take the average of the first pixel values ​​of all the pixel points within the specified area with the pixel point as the center and located within the range of the carrier as the calibration pixel value of the pixel point; According to the calibration pixel value of each pixel point on the reference line and the positional relationship between each pixel point and the reference point, a calibration curve is obtained: A curve fitting is performed on the calibration pixel values ​​of all the pixel points on the reference line and the distance between the corresponding pixel point and the reference point to obtain a calibration curve.

7. The method according to claim 4, characterized in that The set number of pixel points with the highest first pixel values ​​may be selected in the following manner: Grouping the pixel points into a first set, and sorting the pixel points in the first set from large to small according to the first pixel value; Selecting a set number of pixel points that are located at the front when the first set is sorted as pixel points constituting the reference area; The selecting the highest pixel point from the reference area as the reference point comprises: Forming a pixel area with each pixel point in the reference area as the center, wherein one pixel area corresponds to a second pixel value; The pixel areas are sorted according to the second pixel values, starting from the pixel area corresponding to the largest second pixel value, and the central pixel points in the pixel areas are sequentially determined as reference points until the central pixel point is determined as the reference point.

8. The method according to claim 7, characterized in that The forming of a pixel area with each pixel point in the reference area as the center comprises: Taking the same number of adjacent pixel points in multiple directions around the central pixel point, each of the adjacent pixel points and the central pixel point forms a pixel area; The second pixel value is obtained based on the first pixel value, and the obtaining method includes: The sum of the first pixel values ​​corresponding to all the pixel points in the pixel area is calculated, and the sum is selected as the second pixel value.

9. The method according to claim 8, characterized in that The determining of the reference point of the central pixel in the pixel area includes: In multiple directions around the pixel area to be judged, respectively judging whether the pixel values ​​of the same number of pixel points as the pixel points in the pixel area in each direction are less than the first pixel value corresponding to the central pixel point in the pixel area; If the first pixel values ​​corresponding to the central pixel point in the pixel area are all greater than the pixel values ​​of the pixels in all directions, the central pixel point corresponding to the pixel area is determined to be the reference point.

10. The method according to claim 1, characterized in that The confirming the carrier area of ​​the carrier based on the first detection image and the second detection image comprises: Confirming area information of the carrier in the first detection image according to brightness information of the carrier in the first detection image; Determine the fluorescent area in the second detection image according to the brightness information of the fluorescent substance in the second detection image, and confirm the carrier area of ​​the carrier in the second detection image in combination with the area information of the carrier in the first detection image; Confirming the read value of the detected object in the carrier area based on the first detection image and the second detection image includes: The carrier area is divided into a plurality of pixel points, wherein each pixel point corresponds to a pixel value that can reflect area information, and the read value is a second set of pixel values ​​of corresponding pixel points of the detected object in the second detection image.

11. The method according to claim 10, characterized in that Determining a corrected detection value of the read value in the carrier region according to a calibration curve of the read value in the carrier region includes: According to the distance between the pixel points corresponding to all the pixel values ​​of the second set and the reference point in the calibration curve, the corrected pixel value of each pixel point in the carrier area is obtained, and the corrected detection value is a set composed of each of the corrected pixel values.

12. A fluorescence calibration device, applied to an in vitro diagnostic device, wherein the in vitro diagnostic device comprises a carrier for carrying an object to be detected and a camera module for photographing the carrier, characterized in that: include: An acquisition unit, used to acquire a calibration curve and a detection image of the carrier and the detected object taken by the camera module, wherein the detection image includes a first detection image acquired based on a white light source and a second detection image acquired based on a fluorescent light source; A reading unit, configured to confirm a carrier region of the carrier and a reading value of the detected object in the carrier region according to the detection module acquired by the acquisition unit; A calibration unit is used to determine a corrected detection value of the read value in the carrier area according to a calibration curve of the read value in the carrier area.

13. A computer-readable storage medium, characterized in that: The computer-readable storage medium stores a program, and when the program runs, the device controls the computer-readable storage medium to execute the method according to any one of claims 1 to 11.

Images