Digital PCR chip signal reading method
A chip and signal technology, applied in the field of digital PCR chip signal reading, can solve the problem of low detection accuracy and achieve the effect of improving detection accuracy
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Embodiment 1
[0072] Such as figure 1 As shown, the present embodiment provides a digital PCR chip signal reading method, and the digital PCR chip signal reading method specifically includes the following steps:
[0073] Step S1: adding the reaction solution into several reaction chips, oil-sealing each coated reaction chip with mineral oil, and then sealing each reaction chip with a transparent cover. The number of said reaction chips is not less than 2.
[0074] Specifically, in this embodiment, 16 digital PCR reaction chips are selected. The digital PCR chip refers to a planar micropore structure that can separate and fix the PCR reaction system. The micropore structure is a closed hole or a through hole of any geometric shape. The material of the digital PCR chip includes but is not limited to silicon base, Glass, plastic and metal, to name a few.
[0075] Specifically, in this embodiment, the reaction solution is added to each reaction chip in parallel. More specifically, configure...
Embodiment 2
[0113] This embodiment provides a digital PCR chip signal reading method, the digital PCR chip signal reading method is basically the same as in Embodiment 1, the difference is that the FAM fluorescent reagent image is replaced with the VIC fluorescent reagent image to target different detection objects.
[0114] Specifically, in step S3, the acquired fluorescent reagent images include ROX fluorescent reagent images and VIC fluorescent reagent images. Such as Figure 4 Shown is the image of the VIC fluorescent reagent. In this embodiment, the reaction chip is irradiated with light with a wavelength of 450nm-490nm, and the image of the VIC fluorescent reagent is obtained by collecting light with a wavelength of 515nm-530nm by an image sensor.
[0115] Specifically, in step S4, normalization processing is performed on the ROX fluorescent reagent image and the VIC fluorescent reagent image, and the specific steps of the normalization processing are consistent with the first embo...
Embodiment 3
[0121] This embodiment provides a digital PCR chip signal reading method, the digital PCR chip signal reading method is basically the same as the first and second embodiments, the difference is that in step S62, the read gray value is the corresponding coordinate The gray value of the fluorescent reagent image of the pixel and the pixels around the corresponding coordinates.
[0122] Specifically, step S62: read the normalized FAM fluorescent reagent image J′ according to the center coordinates of the ROX positive well position of the corresponding reaction chip acquired in step S5 FAM Or normalized VIC fluorescence reagent image J' VIC The gray value at the corresponding coordinate and the gray value of the coordinates around the corresponding coordinate, and calculate the average value of the gray value. In this embodiment, 9 pixels around the corresponding coordinates are obtained, corresponding to 8 directions of up, down, left, right, upper left, lower left, upper right,...
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