Fluorescence image registration method, gene sequencing instrument and system

Patent metadata

Field	Value
Publication number	US-12039744-B2
Application number	US-202318201309-A
Country	US
Kind code	B2
Filing date	May 24, 2023
Priority date	Apr 10, 2018
Publication date	Jul 16, 2024
Grant date	Jul 16, 2024

Abstract

A fluorescence image registration method includes obtaining at least one fluorescence image of a biochip. An interior local area is selected. Sums of pixel values in the interior local area along a first direction and a second direction are obtained. A plurality of first template lines is selected to find a minimum total value of the sums of pixel values corresponding to the first template lines. Pixel-level correction is performed on a local area of the track line to obtain pixel-level track cross. Other track crosses on the biochip is obtained, and the pixel-level correction is performed on the other track crosses. The position of the pixel-level track line is corrected by a center-of-gravity method to obtain the subpixel-level position of the track line. The subpixel-level positions of all sites uniformly distributed on the biochip is obtained.

First claim

What is claimed is:

1. A fluorescence image registration method applied to a biochip, a pixel distance between track lines on the biochip being set according to template parameter, the fluorescence image registration method comprising:
obtaining at least one fluorescence image of the biochip;
selecting an interior local area of the fluorescence image by selecting an area of 80% of a width along a first direction and 10% of a length along a second direction of the fluorescence image as the interior local area, the first direction being perpendicular to the second direction;
obtaining sums of pixel values in the interior local area of the fluorescence image respectively along the first direction and the second direction, comprising: selecting a plurality of second template lines; moving the plurality of second template lines in the interior local area of the fluorescent image respectively along the first direction and the second direction; and calculating a sum of pixel values in grayscale in the interior local area of the fluorescent image covered by the plurality of second template lines, wherein the sum of pixel values in grayscale is a sum of the gray values of the pixels covered by the plurality of second template lines;
selecting a plurality of first template lines according to the template parameter, traversing the sums of pixel values respectively along the first direction and the second direction by the plurality of first template lines, to find a minimum total value of the sums of pixel values corresponding to the plurality of first template lines respectively along the first direction and the second direction, and a position of the minimum total value along the first direction or the second direction corresponding to a position of the track lines along the first direction or the second direction;
performing pixel-level correction on a local area of the track lines, and a track cross of the track lines after the pixel-level correction being a pixel-level track cross;
obtaining other track crosses on the biochip according to the pixel-level track crosses, and performing the pixel-level correction on the other track crosses;
correcting a position of a pixel-level track line by a center-of-gravity method to obtain a subpixel-level position of the track lines, wherein “correcting the position of the pixel-level track line by the center-of-gravity method” further comprises: selecting an area with a width of 3 pixels and a length of 50 pixels as a local area of the pixel-level track line; obtaining a center of gravity of the local area of the pixel-level track line; and obtaining a subpixel-level position of the track line according to the center of gravity; and
obtaining subpixel-level positions of all sites uniformly distributed on the biochip by equal-size grids dividing method.

2. The fluorescence image registration method of claim 1 , wherein “selecting the plurality of first template lines according to the template parameter, traversing the sums of pixel values respectively along the first direction and the second direction by the plurality of first template lines, to find the minimum total value of the sums of pixel values corresponding to the plurality of first template lines respectively along the first direction and the second direction” further comprises:
calculate a total value of the sums of pixel values respectively along the first direction and the second direction corresponding to the plurality of first template lines; and
obtaining the minimum total value among the total value of the sums of pixel values.

3. The fluorescence image registration method of claim 2 , wherein “performing the pixel-level correction on the local area of the track lines” further comprises:
obtaining the sums of pixel values of the local area of the track line along the first direction and the second direction respectively;
selecting a plurality of third template lines spaced apart from each other by a predetermined distance, to find the sums of pixel values of the local area of the track line by traversal;
obtaining a minimum total value of the sums of pixel values corresponding to the plurality of third template lines; and
obtaining the pixel-level position of the track line according to a position corresponding to the minimum total value.

4. The fluorescence image registration method of claim 3 , wherein “obtaining the pixel-level position of the track line according to the position corresponding to the minimum total value” further comprises:
obtaining a pixel-level position of a valley of W-shaped line according to the position of the minimum total value, wherein the sums of pixel values of the local area of the track line comprises the W-shaped line; and
obtaining the pixel-level position of the track line according to the pixel-level position of the valley.

5. The fluorescence image registration method of claim 4 , wherein the W-shaped line is provided with a peak and two valleys, a pixel distance between one of the two valleys and the peak is constant, and a pixel-level position of the peak is obtained according to the pixel-level position of the valley, and the peak in the W-shaped line corresponds to the track line.

6. The fluorescence image registration method of claim 1 , wherein “obtaining the subpixel-level positions of all sites uniformly distributed on the biochip by equal-size grids dividing method” further comprises:
obtaining a block area formed by the track crosses of two adjacent subpixel-level track lines along the first direction and the second direction, wherein the sites are arranged on the block area according to a preset rule; and
obtaining the subpixel-level position of all of the sites on the block area by the equal-size grids dividing method.

7. A gene sequencing instrument, comprising:
a processor; and
a memory storing one or more computer programs, which when executed by the processor, cause the processor to execute following steps:
obtaining at least one fluorescence image of the biochip;
selecting an interior local area of the fluorescence image by selecting an area of 80% of a width along a first direction and 10% of a length along a second direction of the fluorescence image as the interior local area, the first direction being perpendicular to the second direction;
obtaining sums of pixel values in the interior local area of the fluorescence image respectively along the first direction and the second direction, comprising: selecting a plurality of second template lines; moving the plurality of second template lines in the interior local area of the fluorescent image respectively along the first direction and the second direction; and calculating a sum of pixel values in grayscale in the interior local area of the fluorescent image covered by the plurality of second template lines, the sum of pixel values in grayscale is a sum of the gray values of the pixels covered by the plurality of second template lines;
selecting a plurality of first template lines according to the template parameter, traversing the sums of pixel values respectively along the first direction and the second direction by the plurality of first template lines, to find a minimum total value of the sums of pixel values corresponding to the plurality of first template lines respectively along the first direction and the second direction, and a position of the minimum total value along the first direction or the second direction a position of the track line along the first direction or the second direction;
performing pixel-level correction on a local area of the track lines, and a track cross of the track lines after the pixel-level correction being a pixel-level track cross;
obtaining other track crosses on the biochip according to the pixel-level track crosses,

Assignees

• Mgi Tech Co Ltd

Inventors

• Li Mei
• Li yu-xiang
• Liu Yi-Wen

Classifications

• C12Q2563/107

  fluorescence · CPC title

• G06T2207/30241

  Trajectory · CPC title

• G06T2207/30072

  Microarray; Biochip, DNA array; Well plate · CPC title

• G06T2207/10064

  Fluorescence image · CPC title

• G06T2207/10056

  Microscopic image · CPC title