A method for correcting droplet position between droplet digital pcr channels

A droplet, digital technology, applied in biochemical equipment and methods, microbial determination/inspection, image data processing, etc., can solve problems such as droplet position shift, and achieve the effect of reducing errors and accurate copy number

Active Publication Date: 2021-07-13
PILOT GENE TECH HANGZHOU CO LTD
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] The invention discloses a method for correcting the droplet position between the droplet digital PCR channels, which solves the problem that the droplet position of each channel is offset in the prior art

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  • A method for correcting droplet position between droplet digital pcr channels
  • A method for correcting droplet position between droplet digital pcr channels
  • A method for correcting droplet position between droplet digital pcr channels

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Embodiment 1

[0046] The particularity of droplet digital PCR detection is that the droplet image under one channel is used as the positioning channel to map to the droplet image on other channels, so that the position coordinates of the same droplet under each channel are marked out . The same droplet has different fluorescence value signals under each channel, so each droplet reaction unit corresponds to a multi-dimensional fluorescence data, and it is necessary to correct and analyze the fluorescence crosstalk between channels based on the multi-dimensional fluorescence data. During the actual use of the droplet chip, due to factors such as uneven pressure during the droplet generation process, local temperature unevenness during the PCR amplification process, and changes in light intensity during the use of the PCR reader, the droplets will be interfered to a certain extent. , so that irregular random fretting occurs, such as figure 1 Shown is the trajectory diagram of the random offse...

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Abstract

The invention relates to the field of in vitro DNA amplification detection, in particular to a method for correcting the position of microdrops between microdroplet digital PCR channels. The method for correcting the position of the droplet between the droplet digital PCR channels includes the following steps: S1, feature matching detection: collect the gray scale of the sub-image to be detected and the template image, and calculate the matching degree between the two; S2, direction Feature extraction: perform preliminary correction by extracting directional features; S3, movement trend recognition: further correct the movement trends of all droplets to obtain the corrected droplet position. The method for correcting the position of the droplet between the channels of the droplet digital PCR disclosed by the present invention can accurately locate and accurately match the position of each droplet under each channel, greatly reducing the error of the fluorescence value of each droplet in each channel, The copy number of the detected nucleic acid target molecule is made more accurate, and the method is simple and easy to implement.

Description

technical field [0001] The invention relates to the field of in vitro DNA amplification detection, in particular to a method for correcting the position of microdrops between microdroplet digital PCR channels. Background technique [0002] At present, the digital PCR technology mainly divides the sample into a large number of nanoliter-level micro-reaction units, and then performs PCR cycles on each reaction unit at the same time. After the amplification is completed, the end-point observation method is used to divide each micro-reaction unit into negative or a positive reaction. The traditional solid-state chip is to carve reaction wells with the same shape and size on the substrate. The manufacturing accuracy and the number of reaction wells are limited and cannot meet the optimal progress requirements of digital PCR. Droplet digital PCR generates tens of thousands of droplet microspheres containing the target gene through the microchannel in the droplet chip. For digital...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G06T7/00G06T7/33C12Q1/686
Inventor 程标夏江
Owner PILOT GENE TECH HANGZHOU CO LTD
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