FLUORESCENCE IMAGE REGISTRATION METHOD, GENE SEQUENCING INSTRUMENT AND SYSTEM

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.

Description

Claims

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; andobtaining 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; andobtaining 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; andobtaining 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; andobtaining 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 “correcting the position of the pixel-level track line by the center-of-gravity method” further comprises: obtaining a local area of the pixel-level track line;obtaining a center of gravity of the local area of the pixel-level track line; andobtaining a subpixel-level position of the track line according to the center of gravity.

7. The fluorescence image registration method of claim 6, wherein “obtaining the local area of the pixel-level track line” further comprises: selecting an area with a width of 3 pixels and a length of 50 pixels as the local area of the pixel-level track line.

8. The fluorescence image registration method of claim 6, 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; andobtaining the subpixel-level position of all of the sites on the block area by the equal-size grids dividing method.

9. A gene sequencing instrument, comprising: a processor; anda 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, 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 line; andobtaining subpixel-level positions of all sites uniformly distributed on the biochip by equal-size grids dividing method.

10. The gene sequencing instrument of claim 9, 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; andobtaining the minimum total value among the total value of the sums of pixel values.

11. The gene sequencing instrument of claim 10, wherein “performing the pixel-level correction on the local area of the track lines” 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; andobtaining the pixel-level position of the track line according to a position corresponding to the minimum total value.

12. The gene sequencing instrument of claim 11, 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; andobtaining the pixel-level position of the track line according to the pixel-level position of the valley.

13. The gene sequencing instrument of claim 12, wherein the W-shaped line comprises 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.

14. The gene sequencing instrument of claim 9, wherein “correcting the position of the pixel-level track line by the center-of-gravity method” further comprises: obtaining a local area of the pixel-level track line;obtaining a center of gravity of the local area of the pixel-level track line; andobtaining a subpixel-level position of the track line according to the center of gravity.

15. The gene sequencing instrument of claim 14, wherein “obtaining the local area of the pixel-level track line” further comprises: selecting an area with a width of 3 pixels and a length of 50 pixels as the local area of the pixel-level track line.

16. The gene sequencing instrument of claim 14, 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; andobtaining the subpixel-level position of all the sites on the block area by the equal-size grids dividing method.

17. The gene sequencing instrument of claim 9, wherein the gene sequencing instrument further comprises a chip platform, and an optical system, the chip platform supports the biochip, and the optical system captures the fluorescence images.

18. The gene sequencing instrument of claim 9, wherein the template parameter comprises parameter for designing the biochip.

19. The gene sequencing instrument of claim 9, wherein the plurality of first template lines comprises three first template lines, the sums of pixel values are sequentially searched by the three first template lines.

20. A gene sequencing system applied to a biochip, a pixel distance between track lines on the biochip is set according to template parameter, the gene sequencing system comprising: an image obtaining module, configured to obtain at least one fluorescence image of the biochip;an area selecting module, configured to select 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 is perpendicular to the second direction;a sum of pixel values obtaining module, configured to obtain sums of pixel values in the interior local area of the fluorescence image respectively along the first direction and the second direction, and further configured to executed following functions: 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;a minimum total value finding module, configured to select a plurality of first template lines according to the template parameter, traverse 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;a pixel-level correcting module, configured to perform pixel-level correction on a local area of the track lines, and a track cross of the track lines after the pixel-level correction is a pixel-level track cross;an other track cross obtaining module, configured to obtain other track crosses on the biochip according to the pixel-level track crosses, and perform the pixel-level correction on the other track crosses;a center-of-gravity correcting module, configured to correct a position of a pixel-level track line by a center-of-gravity method to obtain a subpixel-level position of the track line; anda subpixel-level site obtaining module, configured to obtaining subpixel-level positions of all sites uniformly distributed on the biochip by equal-size grids dividing method.