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DPCR (Digital Polymerase Chain Reaction)-based novel full-automatic fluorescent signal acquisition and analysis method

A fluorescence signal, acquisition and analysis technology, applied in the field of image processing, can solve the problems of reducing detection efficiency, increasing operational complexity, unfavorable system integration development, etc., to avoid interference, improve accuracy and detection efficiency.

Active Publication Date: 2018-02-23
NORTHWESTERN POLYTECHNICAL UNIV
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Problems solved by technology

Although the final result is obtained, manual marking increases the complexity of the operation, which is not conducive to the integrated development of the system, and progressive scanning reduces the detection efficiency

Method used

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  • DPCR (Digital Polymerase Chain Reaction)-based novel full-automatic fluorescent signal acquisition and analysis method
  • DPCR (Digital Polymerase Chain Reaction)-based novel full-automatic fluorescent signal acquisition and analysis method
  • DPCR (Digital Polymerase Chain Reaction)-based novel full-automatic fluorescent signal acquisition and analysis method

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Embodiment Construction

[0040] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0041] The steps of a novel automatic fluorescent signal acquisition and analysis method based on dPCR are as follows:

[0042] Step 1: Fabrication of the dPCR chip. We have produced high-density microwell dPCR chips with special markers by using micro-nano processing technology, such as Figure 10 : The overall size is 9.1*9.1㎜, the edge size of the middle micro-reaction chamber is 8380*8380μm, and there are six special cross marks in the ring-shaped area formed by the edge of the dPCR chip and the edge of the micro-reaction chamber A, marked as P1 , P2, P3, P4, P5, P6, it is stipulated that the direction connecting the center points of P1 and P2 and pointing to P2 is the positive direction of x, and the direction connecting the center points of P1 and P5 and pointing to P5 is the positive direction of y. The center coordinate is P1(200,200) (unit is μm), ...

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Abstract

The invention belongs to the field of image processing and in particular discloses a dPCR (Digital Polymerase Chain Reaction)-based novel full-automatic fluorescent signal acquisition and analysis method. According to the method, a fluorescent signal is extracted and analyzed by utilizing an idea of image region segmentation-image stitching-signal acquisition and analysis. Due to the method, the fluorescent signal acquisition and analysis method is applied to result detection at the later stage of dPCR in an engineered manner firstly, and due to a reasonable design, full automation of the whole system is realized, measurement errors caused by artificial interference such as manual marking, progressive scanning and the like are avoided, the accuracy is high, the operation and control are convenient, and the detection precision and the detection efficiency of the system are effectively improved. Due to the stitching step, the acquired scattered images can be perfectly stitched to be of the size of the original image during designing, the original image is highly reduced, convenience is brought to fluorescent signal analysis and detection at the later stage, and the detection accuracyis improved.

Description

technical field [0001] The invention belongs to the field of image processing, in particular to a novel automatic fluorescent signal acquisition and analysis method based on dPCR (digital Polymerase Chain Reaction). Background technique [0002] The concept of dPCR technology has developed rapidly in less than 20 years since it was put forward. This technology amplifies genes in vitro, and is a molecular biology technique used to amplify specific DNA fragments. At present, as a powerful tool for absolute quantitative detection of target genes, transgenic detection, genetic defect screening, cancer prevention, gene mutation detection, etc., dPCR technology has extremely wide applications in biomedicine and precision medicine. However, most of the current research focuses on optimizing and improving the dPCR chip itself, while the detection of late-stage fluorescent signals in the entire dPCR system has become a "gray area" of this major research. [0003] After searching th...

Claims

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Application Information

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IPC IPC(8): C12Q1/686G01N21/64
CPCC12Q1/686G01N21/64C12Q2563/159C12Q2563/107
Inventor 帕维尔·诺伊茨尔李桦楠徐颖张浩卿
Owner NORTHWESTERN POLYTECHNICAL UNIV
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