Method for detecting hybridization result of gene chip through total internal reflection

A technology for detecting gene chips and total internal reflection, which is applied in the field of nucleic acid analysis, can solve problems such as false positives and false negatives, and achieve the effect of improving signal-to-noise ratio, solving background interference, and good use effect

Active Publication Date: 2021-06-15
BEIJING BIONAXIN BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present disclosure is also to provide a method for detecting gene chip hybridization results by total internal reflection, which overcomes the problem of false negatives or false positives caused by background interference of excitation light on fluorescence imaging results

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  • Method for detecting hybridization result of gene chip through total internal reflection
  • Method for detecting hybridization result of gene chip through total internal reflection

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specific Embodiment 1

[0040] Specific embodiment 1: according to figure 2 After the detector is constructed as shown, the equal refractive index solution 9 is added dropwise on the quartz glass plate 2. Here, the equal refractive index solution 9 we choose is an ethylene glycol solution with the same refractive index as the quartz glass, and the A blank quartz glass substrate 10 is placed on the top of the equal refractive index solution 9 to form an integrated structure of "quartz glass plate-ethylene glycol solution-blank quartz glass substrate" with a consistent refractive index. Then, the blank quartz glass substrate 10 Add dropwise the nucleic acid solution containing AlexaFluor 488 labeled (prepared by introducing EdU (thymidine (thymidine) analogue, 5-Ethynyl-2'-deoxyuridine) by PCR method), turn on the plane parallel light source generator 6, Here, the plane-parallel light source generator 6 includes a laser light source and a laser beam expander. The product selects the MDL-XS-488 model o...

specific Embodiment 2

[0041] Specific embodiment 2: after completing the debugging of the optical path system according to specific embodiment 1, take out the blank quartz glass substrate 10, ensure that the ethylene glycol solution of the equal refractive index solution 9 is still full, and complete the hybridization and fluorescently labeled quartz glass The substrate 10 is placed on the upper part of the medium refractive index solution 9 in the imaging dark box 16; the surface parallel light source generator 6 is turned on, and the fluorescent image of the hybridization result of the gene chip is obtained by the high-resolution camera 15, and the obtained fluorescent imaging image is uploaded to the data center Analysis of gene chip hybridization detection results.

[0042] Through specific embodiment 1, the position and angle of the plane-parallel light source generator 6 are adjusted to ensure that the excitation light 7 of the light source forms the evanescent wave 11 of total internal reflec...

specific Embodiment 3

[0044] Specific embodiment 3: In the process of constructing a total internal reflection type gene chip detector, in order to ensure that the light source excitation light irradiated on the substrate of the gene chip by plane parallel light can irradiate the substrate covering the entire gene chip, such as the gene chip The substrate is made of quartz glass substrate 10. There may be part of the light source excitation light 7 passing through the two ends of the quartz glass substrate 10. Set the movable blocking light reflective material 17. After the quartz glass substrate 10 is placed, adjust the movable blocking light reflective material 17 to ensure that the two ends of the quartz glass substrate 10 are closely connected to the movable blocking light reflective material 17. are connected to further ensure the background interference of the excitation light 7 of the light source to the imaging.

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Abstract

The invention discloses a method for detecting gene chip hybridization results through total internal reflection, which comprises the following steps of constructing a gene chip detector, and dividing a camera obscura (1) of the gene chip detector into a light source camera obscura (3) and an imaging camera obscura (16) by using a quartz glass plate (2); adding an equal refractive index solution (9) with the same refractive index as the quartz glass on the quartz glass plate (2); placing a quartz glass substrate of the gene chip on an equal refractive index solution (9); adjusting the position and an angle of the plane parallel light source generator (6) so that the emitted plane parallel light forms total internal reflection; the total internal reflection evanescent wave (11) exciting a fluorescent substance on a probe (12) of the gene chip to emit marking fluorescence (13); and the labeled fluorescence (13) being captured by a high-resolution camera (15), and finally a detection result being obtained. According to the method, background interference in the fluorescence imaging process is avoided, and the signal-to-noise ratio and detection sensitivity of a gene chip hybridization result are improved.

Description

technical field [0001] The invention relates to the technical field of nucleic acid analysis, which is a method for determining nucleic acid and adopts a product for nucleic acid analysis. The invention particularly relates to a method for detecting the hybridization result of a gene chip by total internal reflection. The invention also particularly relates to an in vitro diagnostic testing instrument, which belongs to the high-throughput testing and analysis instrument in the in vitro diagnostic testing instrument, and also relates to a molecular biological information analysis and processing system in the molecular diagnostic testing instrument. Background technique [0002] Biochip technology has been widely used in clinical disease diagnosis, health management, drug research and development, animal and plant quarantine, food testing, environmental monitoring, scientific research, forensic testing and many other fields. It has broad application prospects and a very large m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N21/01
CPCG01N21/01G01N21/6402G01N21/6428G01N21/6456G01N2021/6463G01N2021/6471
Inventor 华子昂刘宝全竹添朱美瑛万君兴张建李娜
Owner BEIJING BIONAXIN BIOTECH CO LTD
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