CN-121994457-A - Imaging quality evaluation method and related device

CN121994457ACN 121994457 ACN121994457 ACN 121994457ACN-121994457-A

Abstract

The application discloses an imaging quality evaluation method and device. The method comprises the steps of obtaining image sequences acquired by an imaging device in different view field areas of a calibration plate and serving as evaluation image sequences for evaluating imaging quality of the imaging device, dividing evaluation images in the evaluation image sequences into a plurality of image blocks respectively, determining full widths of half peaks of the image blocks, determining an inclination angle of an optical axis of the imaging device and the calibration plate based on the full widths of the half peaks of the image blocks divided by the evaluation images in the evaluation image sequences, calculating target full widths of half peaks of the image blocks in a view field of the imaging device based on the inclination angle of the optical axis of the imaging device and the calibration plate and the full widths of the half peaks of the image blocks divided by the evaluation images in the evaluation image sequences, and determining imaging quality of the imaging device based on the target full widths of half peaks of the image blocks in the view field of the imaging device. The application can conveniently and rapidly evaluate the imaging quality, improve the accuracy and the accuracy of the evaluation result and reduce the evaluation cost.

Inventors

CAI KEYA
QIAO SHUQI
FANG ZHENG
WANG LIANG
ZHANG JIABEI
HOU XIZHE

Assignees

郑州思昆生物工程有限公司

Dates

Publication Date: 20260508
Application Date: 20260121

Claims (11)

1. An imaging quality evaluation method, characterized by comprising: Acquiring image sequences acquired by an imaging device in different view field areas of a calibration plate and serving as evaluation image sequences for evaluating imaging quality of the imaging device, wherein the calibration plate comprises micropores which are arranged in an array, luminescent substances are filled in the micropores for imaging of the calibration plate, and the micropores form light spots after imaging; dividing an evaluation image in the evaluation image sequence into a plurality of image blocks respectively, and determining the full width at half maximum of the image blocks; Determining an inclination angle of an optical axis of the imaging device and the calibration plate based on the full width at half maximum of an image block divided by an evaluation image in the evaluation image sequence; Calculating the target full width at half maximum of each image block in the field of view of the imaging device based on the inclination angle of the optical axis of the imaging device and the calibration plate and the full width at half maximum of the image block divided by the evaluation image in the evaluation image sequence; The imaging quality of the imaging device is determined based on the target full width at half maximum of each image block within the field of view of the imaging device.
2. The method according to claim 1, wherein acquiring the image sequences acquired by the imaging device at different field of view areas of the calibration plate and serving as an evaluation image sequence for evaluating the imaging quality of the imaging device comprises: Acquiring a first image sequence acquired by an imaging device in different view field areas of a calibration plate; Determining a focusing position of the imaging device according to the definition of the images in the first image sequence; acquiring a second image sequence acquired by the imaging device at a plurality of defocusing positions in the focusing position and/or the defocusing region and taking the second image sequence as the evaluation image sequence; the out-of-focus area is centered on the focusing position, and the distance between the edge position of the out-of-focus area and the focusing position is preset.
3. The method of claim 1, wherein said determining the full width half maximum of the image block comprises: determining the center of a light spot in the image block based on the gray value of a pixel point in the light spot in the image block; Calculating the full width at half maximum of the light spot based on the center of the light spot; and determining the full width half maximum of the image block based on the full width half maximum of the light spot in the image block.
4. A method according to claim 3, wherein said calculating the full width at half maximum of the spot based on the center of the spot comprises: Calculating a plurality of target constant-integration values of two-dimensional Gaussian distribution based on gray values of pixel points in a plurality of distribution areas where the centers of the light spots are located, wherein the sizes of the distribution areas are preset; respectively performing polynomial fitting on the constant integration values corresponding to different standard deviations in a plurality of preset constant integration ranges to obtain polynomials corresponding to the constant integration ranges; Calculating standard deviation corresponding to each target constant-integration value of the two-dimensional Gaussian distribution based on polynomials corresponding to each constant-integration range; and calculating the full width at half maximum of the light spot based on standard deviation corresponding to each target fixed and integrated value of the two-dimensional Gaussian distribution.
5. The method according to claim 1, wherein the determining the tilt angle of the optical axis of the imaging device to the calibration plate based on the full width at half maximum of the image block divided by the evaluation image in the sequence of evaluation images comprises: determining image blocks with minimum half-width at different positions of the divided image blocks based on the half-width of the image blocks divided by the evaluation images in the evaluation image sequence; Constructing a first height matrix according to the Z-axis height of the imaging device when the evaluation image of the image block with the minimum half-width at the position of different image blocks is acquired; and determining an inclination angle of an optical axis of the imaging device and the calibration plate based on the first height matrix.
6. The method of claim 5, wherein determining an inclination angle of an optical axis of the imaging device with the calibration plate based on the first height matrix comprises: Performing linear fitting on the row vectors in the first height matrix and the lengths of the corresponding imaging device visual fields in the X-axis direction to obtain a first linear slope; calculating an inclination angle of an optical axis of the imaging device with the calibration plate in an X-axis direction based on the first linear slope; performing linear fitting on column vectors in the first height matrix and the width of the corresponding imaging device view field in the Y-axis direction to obtain a second linear slope; calculating an inclination angle of an optical axis of the imaging device and the calibration plate in a Y-axis direction based on the second linear slope; Wherein the length of the field of view in the X-axis direction and the width of the field of view in the Y-axis direction are each determined by the dimensions of the evaluation image and the image block.
7. The method of claim 1, wherein calculating the target full-width-half-maximum for the image block within the field of view of the imaging device based on the tilt angle of the optical axis of the imaging device to the calibration plate, the full-width-half-maximum for the image block divided by the evaluation image in the sequence of evaluation images, comprises: Determining a plane equation of a target focal plane based on an inclination angle of an optical axis of the imaging device and the calibration plate; determining the Z-axis height on the target focal plane corresponding to each image block based on a plane equation of the target focal plane; Calculating the target full width at half maximum of the image blocks in the field of view of the imaging device based on the full width at half maximum of the image blocks divided by the evaluation images in the evaluation image sequence and the Z-axis height on the target focal plane corresponding to each image block; wherein a focus position of the imaging device is located on the target focal plane.
8. The method of claim 7, wherein calculating the target full width half maximum for each image block within the field of view of the imaging device based on the full width at half maximum of the image block divided by the evaluation image in the sequence of evaluation images, the Z-axis height at the target focal plane for each image block, comprises: and performing linear interpolation based on the Z-axis height of each evaluation image during imaging, the full width at half maximum of the image block divided by the evaluation image and the Z-axis height on the target focal plane corresponding to each image block to obtain the target full width at half maximum of each image block in the field of view of the imaging device.
9. A computer device, comprising: At least one processor, and A memory communicatively coupled to the at least one processor, wherein, The memory stores instructions executable by the at least one processor to cause the at least one processor to implement the method of any one of claims 1-8.
10. A gene sequencer, comprising: The calibration plate comprises micropores which are arranged in an array, fluorescent substances are filled in the micropores for imaging of the calibration plate, and the micropores form light spots after imaging; A chip stage configured to fix and support the calibration plate; The electronic control system is configured to control the relative movement between the chip platform and the optical detection system in the X-axis direction and the Y-axis direction so that the optical detection system can acquire fluorescent images of different view field areas on the calibration plate; an optical detection system configured to excite a fluorescent substance in the calibration plate and collect a fluorescent signal, and generate a fluorescent image according to the fluorescent signal and serve as an evaluation image for evaluating the quality of the imaging device; a computer system comprising at least one processor and a memory communicatively coupled to the at least one processor, wherein the memory stores instructions executable by the at least one processor to cause the at least one processor to implement the method of any of claims 1-8.
11. The gene sequencer according to claim 10, further comprising: A sequencing chip configured to provide a reaction region for an amplification reaction, a sequencing reaction; A reagent storage container configured to store one or more mixed sample libraries, one or more reagents; A fluidic system configured to controllably deliver one or more mixed sample libraries, one or more reagents from the reagent storage container into the sequencing chip for performing amplification reactions, sequencing reactions in the sequencing chip, and controllably delivering reacted waste liquid from the sequencing chip into the waste liquid storage container; A waste liquid storage container configured to store waste liquid generated after the reaction; the optical detection system is further configured to excite a fluorescent group and collect a fluorescent signal during a sequencing reaction, and generate a fluorescent image according to the fluorescent signal; a computer system further configured to acquire a fluorescence image from the optical detection system and identify a base sequence of the sample library from the fluorescence image; The electric control system is also configured to control the operation and the work of the liquid path system and the optical detection system.

Description

Imaging quality evaluation method and related device Technical Field The present application relates to the field of image processing technologies, and in particular, to an imaging quality evaluation method and a related apparatus. Background In the field of gene sequencing, a mainstream gene sequencer is to acquire a fluorescent image on a sequencing chip through an imaging device and analyze a fluorescent signal in the fluorescent image to obtain a gene sequence. In the optical system, the high-quality imaging device can generate clear, sharp and high-contrast images, and for the gene sequencer, the quality of the imaging device determines the quality of fluorescent images and further determines the quality of gene sequencing results, so that the performance of the imaging device can be accurately measured through imaging quality evaluation. The prior art generally detects an imaging device based on a wavefront sensing device and an MTF (Modulation Transfer Function ) sensing device to determine its imaging quality. But the detection equipment needs to be accurately calibrated and adjusted every time, and the imaging device can only be installed on the detection equipment for detection, so that the evaluation process is complicated, the cost is high, and the evaluation result is easily influenced by the experience of operators. Disclosure of Invention The application provides an imaging quality evaluation method and device, which can conveniently and rapidly evaluate the imaging quality, improve the accuracy and the accuracy of an evaluation result and reduce the evaluation cost. In a first aspect, an embodiment of the present application provides an imaging quality evaluation method, including: Acquiring image sequences acquired by an imaging device in different view field areas of a calibration plate and serving as evaluation image sequences for evaluating imaging quality of the imaging device, wherein the calibration plate comprises micropores which are arranged in an array, luminescent substances are filled in the micropores for imaging of the calibration plate, and the micropores form light spots after imaging; dividing the evaluation image in the sequence of evaluation images into a plurality of image blocks, And determining a full width at half maximum of the image block; Determining an inclination angle of an optical axis of the imaging device and the calibration plate based on the full width at half maximum of an image block divided by an evaluation image in the evaluation image sequence; Calculating the target full width at half maximum of each image block in the field of view of the imaging device based on the inclination angle of the optical axis of the imaging device and the calibration plate and the full width at half maximum of the image block divided by the evaluation image in the evaluation image sequence; The imaging quality of the imaging device is determined based on the target full width at half maximum of each image block within the field of view of the imaging device. According to a second aspect of the present application, there is provided a computer device comprising at least one processor, and a memory communicatively coupled to the at least one processor, wherein the memory stores instructions executable by the at least one processor to cause the at least one processor to implement an imaging quality assessment method according to any of the embodiments of the present application. According to a third aspect of the present application, there is provided a gene sequencer comprising: The calibration plate comprises micropores which are arranged in an array, fluorescent substances are filled in the micropores for imaging of the calibration plate, and the micropores form light spots after imaging; A chip stage configured to fix and support the calibration plate; The electronic control system is configured to control the relative movement between the chip platform and the optical detection system in the X-axis direction and the Y-axis direction so that the optical detection system can acquire fluorescent images of different view field areas on the calibration plate; an optical detection system configured to excite a fluorescent substance in the calibration plate and collect a fluorescent signal, and generate a fluorescent image according to the fluorescent signal and serve as an evaluation image for evaluating the quality of the imaging device; The computer system comprises at least one processor and a memory communicatively connected to the at least one processor, wherein the memory stores instructions executable by the at least one processor to cause the at least one processor to implement the method as recited in any of the preceding claims. According to the imaging quality evaluation method and the related device, the full width at half maximum is corrected by calculating the inclination angle of the optical axis and the calibration plate, so that systematic errors caused by ins