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Fluorescence image registration method

A fluorescence image and image registration technology, applied in the field of sequencing fluorescence image registration, can solve problems such as difficulty in finding and large amount of data, and achieve the effect of simple algorithm process, small calculation amount, and strong robustness.

Pending Publication Date: 2022-05-27
CYGNUS BIOSCI BEIJING CO LTD
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Problems solved by technology

During high-throughput sequencing, the sequencing chip moves relative to the image acquisition components such as the objective lens. Due to the movement error of the translation stage, the position of the bright spot corresponding to the same microwell in the fluorescence image of each sequencing cycle will have a slight deviation, and the high The data volume of throughput sequencing is relatively large, and the general gene sequencing chip has a periodic structure or distribution, so it is difficult to find the DNA molecule at the same position during multiple sequencing

Method used

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Embodiment 1

[0059] 1. Sequence the gene sequence to be measured on the gene sequencing chip, and obtain a sequenced fluorescence image with a width*height, that is, a wide pixel multiplied by a high pixel.

[0060] 2. Select the fluorescent images taken at each preset camera position in the first round of sequencing reaction as the template image, and the remaining images at the same location as the images to be registered.

[0061] 3. For each sequenced fluorescence image, divide into t(t greater than or equal to 1) sub-regions by row, such as Figure 1A Shown is a sequencing image, there are horizontal M = 3 positioning lines in the figure, longitudinal N = 3 positioning lines, in the present embodiment, the image is divided into t = 8 sub-regions by column (i.e., in the vertical direction) to extract the positioning line features, for each sub-region, the following calculations:

[0062] Calculate the sum of the pixels of each column to obtain a floating-point (integer) matrix of 1 row of w...

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Abstract

The invention discloses a fluorescence image registration method, and belongs to the field of gene sequencing. And carrying out image registration through a preset positioning line. According to the fluorescent image registration method, image registration can be completed by identifying fluorescent image characteristic positioning marks and comparing position differences of the fluorescent image characteristic positioning marks in different sequencing cycles depending on the positioning lines which are preset on the gene sequencing chip and distributed at equal intervals.

Description

Technical field [0001] The present invention relates to a fluorescence image registration method, belonging to the field of gene sequencing, specifically, relates to a sequencing fluorescence image registration method. Background [0002] The process of sequencing genes generates a large amount of data. In general, second-generation sequencing uses signals generated during chemical reactions to detect bases. When detecting, the signal of a single DNA molecule is very weak and it is difficult to detect, so it is necessary to copy a single DNA molecule or fragment to a large number, such as 210, by amplification, etc., to facilitate the detection of the signal. During the assay, each chemical reaction extends the complementary strands of the DNA strand to be measured by 1 or more bases. Depending on the sequencing method, for example, the 3-terminal blocked Illumina-like sequencing method extends 1 base at a time; for example, a 2+2 type sequencing method may extend multiple bases ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G06T7/33G16B20/30
CPCG06T7/33G16B20/30G06T2207/10064
Inventor 李绍森周文雄李昂乔朔
Owner CYGNUS BIOSCI BEIJING CO LTD
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