Inter-lab difference adjustment method for microbial automatic microscope machine

Abstract

The application discloses a kind of interbench difference debugging method of microbial automatic microscope machine, comprising: ①preparing standard sample ②placing standard sample below objective lens, and make the bacterial sample on sample appear in objective lens observation area range ③make camera automatic focusing ④keep camera system focal length unchanged, move standard sample, and make blank area of cover glass be located in objective lens observation area range ⑤select brightness control area, and according to nine grid method, calculate real-time brightness value, adjust camera exposure time, so that the difference between real-time brightness value and standard brightness value is within ±5 ⑥determine color control area, according to nine grid method, calculate real-time color value, adjust camera color temperature setting, TINT setting, so that the difference between real-time color value and standard color value is within ±5 ⑦save setting, complete interbench difference debugging.The application proposes nine palace format to divide brightness, color area, optimizes brightness, color calculation method, effectively controls machine interbench difference.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of microorganism detection equipment, in particular to a method for debugging inter-table difference of a microorganism automatic microscope machine. BACKGROUND

[0002] A traditional optical microscope needs to be observed by naked eyes, which is time-consuming and laborious, requires high professional and technical quality of the workers, and takes a long time, and the probability of subjective misjudgment is high. With the development of science and technology, automatic microscope machines that can convert optical signals into digital signals have gradually been popularized. When in use, the sample only needs to be placed in the observation area of the automatic microscope machine, and the automatic microscope machine can automatically take pictures, identify and output detection results, and the detection speed and accuracy of the detection results are greatly improved. Since the detection of the automatic microscope machine is mainly based on the pictures taken by the camera, it is crucial for the equipment quality control to make the image color and brightness of the microscope camera meet the requirements. If the inter-table difference of the automatic microscope machine cannot be strictly controlled before it is shipped, the photographed images of different automatic microscope machines for the same sample may have color and brightness differences, which will eventually affect the interpretation results and cause misdiagnosis of the patient. The existing inter-table difference debugging method of the microscope is mainly for traditional optical microscopes, which is low in efficiency and not suitable for the above-mentioned automatic microscope instruments. SUMMARY

[0003] To solve the above problems, the present application provides a method for debugging inter-table difference of a microorganism automatic microscope machine, which can specifically adopt the following technical solutions:

[0004] The method for debugging inter-table difference of the microorganism automatic microscope machine comprises the following steps:

[0005] Firstly, a standard sample is prepared, which is consistent in size with a standard microscope slide, and a bacterial sample dyed by Gram is arranged on the standard sample, and the bacterial sample is packaged by a cover glass;

[0006] Secondly, the standard sample is placed below the objective lens, and the bacterial sample is made to appear in the objective lens observation area;

[0007] Thirdly, the camera is automatically focused to obtain a clear bacterial pattern;

[0008] Fourthly, the focal length of the camera system is kept unchanged, the standard sample is moved, the bacterial pattern on the standard sample is moved out of the objective lens observation area, and the blank area of the cover glass is made to be located in the objective lens observation area;

[0009] In the fifth step, a brightness control area is selected in the objective observation area, and a real-time brightness value is calculated according to the nine-square method, which is compared with a standard brightness value, and the camera exposure time is adjusted so that the difference between the real-time brightness value and the standard brightness value is within ±5;

[0010] In the sixth step, a color control area is determined according to the brightness control area selected in the fifth step, a real-time color value is calculated according to the nine-square method, which is compared with a standard color value, and the camera color temperature setting and TINT setting are adjusted so that the difference between the real-time color value and the standard color value is within ±5;

[0011] In the seventh step, the settings are saved, and the inter-stage difference adjustment is completed.

[0012] The nine-square method for calculating the real-time brightness value in the fifth step includes: dividing the brightness control area into nine squares according to three rows and three columns, selecting nine pixel points in each square, reading out the brightness values of the nine pixel points, respectively, then calculating the average value, which is the real-time brightness value of the square, and comparing the real-time brightness value of each square with the standard brightness value.

[0013] The camera exposure time in the fifth step is 0.5-5 ms.

[0014] The nine-square method for calculating the real-time color value in the sixth step includes: taking the brightness control area as the color control area, and dividing it into nine squares according to three rows and three columns, selecting nine pixel points in each square, reading out the RGB values of the nine pixel points, respectively, then calculating the average value of RGB, which is the real-time color value of the square, and comparing the real-time color value of each square with the standard color value.

[0015] In the fifth step and the sixth step, if the real-time brightness values and the real-time color values of the nine squares differ too much, the light source of the microscope machine and / or the inclination of the objective lens can be adjusted.

[0016] The inter-stage difference adjustment method of the automatic microbial microscope machine provided by the application can solve the problems of image color and brightness difference that may occur when different microscope machines take pictures, and proposes a nine-square format to divide brightness and color areas, optimizes the brightness and color calculation method, standardizes, speeds up and accurately compares the brightness and color, effectively controls the inter-stage difference of the automatic microbial microscope machine, makes the inter-stage difference (mainly including microscopic image color and brightness) of the factory automatic microbial microscope machine approximately consistent, keeps it within a limited range, and avoids the misdiagnosis phenomenon caused by the brightness and color of the photographed image. BRIEF DESCRIPTION OF DRAWINGS

[0017] Fig. 1 is a debugging step block diagram of the application.

[0018] Fig. 2 is the clear image of the focus of the strain sample in the objective observation area in the present application.

[0019] Fig. 3 is the image of the blank area of the cover glass divided according to the nine-square method (including real-time brightness and color value) in the present application. DETAILED DESCRIPTION

[0020] The embodiments of the present application will be described in detail below with reference to the accompanying drawings, which are implemented on the premise of the technical scheme of the present application, and detailed implementation manner and specific step method are given, but the protection scope of the present application is not limited to the following embodiments.

[0021] As shown in Figs. 1-3 , the inter-bench difference debugging method of the automatic microscope machine for microorganisms comprises the following steps:

[0022] Firstly, a standard sample is prepared, which is consistent in size with a standard microscope slide, and carries a strain sample dyed by Gram, and the strain sample is packaged by a cover glass. For example, a standard sample piece of Rainforest brand (75mmx25mmx1.1mm) can be selected, which is consistent in size with a standard microscope slide, and the strain sample dyed by Gram is packaged by a cover glass on the slide, and the strain sample can be fungi, coccus, Escherichia coli, Candida albicans and the like; the cover glass conforms to the national standard, and the material is glass with a thickness of 0.17±0.01mm.

[0023] Secondly, a standard sample is taken and placed on the objective table of the microscope, and the strain sample is located directly below the objective, i.e. the strain sample is in the objective observation area.

[0024] Thirdly, the camera is automatically focused to obtain a clear strain pattern (see Fig. 2 ).

[0025] Fourthly, after the automatic focusing is completed, the focal length of the camera system is kept unchanged, the standard sample is moved, and the strain pattern on the standard sample is moved out of the objective observation area, while the blank area of the cover glass is located in the objective observation area; at this time, although there is no strain sample between the slide and the cover glass, the image can still be clear due to the unchanged focus position.

[0026] Fifthly, the blank area of the cover glass in the objective observation area is taken as a brightness control area, and the real-time brightness value is calculated according to the nine-square method, which is compared with the standard brightness value, the exposure time of the camera is adjusted, so that the difference between the real-time brightness value and the standard brightness value is within ±5.

[0027] Specifically, the nine-square method calculates the real-time brightness value. First, the brightness control area within the field of view is divided into nine squares according to three rows and three columns. Nine pixel points are selected in each square. The brightness values of the nine pixel points are read out, and the average value is calculated, which is the real-time brightness value of the square. The real-time brightness value of each square is compared with the standard brightness value. The partition method and the brightness calculation method of the nine-square format can avoid data deviation caused by uneven value area and avoid the failure of brightness adjustment caused by the actual brightness being uneven but the numerical value being uniform.

[0028] The exposure time of the camera is usually adjusted in the range of 0.5-5 ms.

[0029] In the sixth step, the color control area is determined according to the brightness control area selected in step five. The real-time color value is calculated according to the nine-square method, which is compared with the standard color value. The camera color temperature setting and TINT setting are adjusted so that the difference between the real-time color value and the standard color value is within ±5.

[0030] Specifically, the brightness control area is used as the color control area, and the real-time color value is calculated according to the nine-square method. First, the color control area within the field of view is divided into nine squares according to three rows and three columns, which can avoid the situation that the color of the image center and the edge is different and cannot be detected. Then, nine pixel points are selected in each square. The RGB values of the nine pixel points are read out, and the average value of RGB is calculated, which is the real-time color value of the square. The real-time color value of each square is compared with the standard color value.

[0031] The data acquisition method of selecting nine pixel points in each square can avoid false judgment caused by too large difference in final brightness or RGB color due to dust on the surface of the camera pixels.

[0032] In addition, during the adjustment process, if the real-time brightness value and the real-time color value of the nine squares are too different, the light source and / or the inclination of the objective lens of the microscope machine can be adjusted.

[0033] The standard brightness value and the standard color value used in steps five and six come from the brightness value and the RGB color value of a standard photo. The standard photo is taken by a designated automatic microscope machine. The color and brightness of the equipment are adjusted. After the brightness and color of the photo taken by the equipment are confirmed to be qualified by professionals, the brightness and color of the photo are determined as the standard brightness value and the standard color value.

[0034] In the seventh step, the focus parameter setting, the exposure time parameter setting, the camera color temperature setting, and the TINT setting are saved, and the inter-machine difference adjustment is completed.

[0035] It should be noted that in the description of the present application, the terms indicating the orientation or positional relationship such as "front", "back", "left", "right", "vertical", "horizontal", "inner", "outer" and the like are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the present application.

Claims

1. A method for adjusting the inter-stage difference of an automated microbial microscopy machine, characterized in that, Includes the following steps: The first step is to prepare a standard sample slide, which is the same size as a standard microscope slide. The standard sample slide contains a Gram-stained bacterial sample, and the bacterial sample is sealed with a coverslip. The second step is to place the standard sample under the objective lens and ensure that the bacterial sample appears within the objective lens observation area; The third step is to autofocus the camera to obtain a clear image of the fungal strain. Fourth step: Keep the camera system focal length unchanged, move the standard sample to move the bacterial pattern on the standard sample out of the objective lens observation area, and at the same time make the blank area of ​​the coverslip within the objective lens observation area. Fifth, select a brightness control area within the objective lens observation area, calculate the real-time brightness value using the nine-square grid method, compare it with the standard brightness value, and adjust the camera exposure time so that the difference between the real-time brightness value and the standard brightness value is within ±5. Step 6: Determine the color control area based on the brightness control area selected in step 5, calculate the real-time color value using the nine-grid method, compare it with the standard color value, and adjust the camera color temperature setting and TINT setting so that the difference between the real-time color value and the standard color value is within ±5. Step 7: Save the settings and complete the inter-station differential adjustment.

2. The method for adjusting the inter-stage difference of the automated microbial microscopy machine according to claim 1, characterized in that: The real-time brightness value calculation using the nine-grid method in step five includes: dividing the brightness control area into nine squares in three rows and three columns; selecting nine pixels in each square; reading the brightness values ​​of the nine pixels; calculating the average value, which is the real-time brightness value of the square; and comparing the real-time brightness value of each square with the standard brightness value.

3. The method for adjusting the inter-stage difference of the automated microbial microscopy machine according to claim 2, characterized in that: The camera exposure time in step five is 0.5-5ms.

4. The method for adjusting the inter-stage difference of the automated microbial microscopy machine according to claim 3, characterized in that: The real-time color value calculation using the nine-grid method in step six includes: taking the brightness control area as the color control area, dividing it into nine squares in three rows and three columns, selecting nine pixels in each square, reading the RGB values ​​of the nine pixels, and then calculating the average RGB value, which is the real-time color value of the square. The real-time color value of each square is then compared with the standard color value.

5. The method for adjusting the inter-stage difference of the automated microbial microscopy machine according to claim 4, characterized in that: In steps five and six, if the real-time brightness and color values ​​of the nine squares differ too much, they can be adjusted by adjusting the light source and / or the tilt of the objective lens of the microscopic examination machine.

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