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Solid phase carrier chip system for in-situ hybridization and application thereof

A technology of solid-phase carrier and chip system, which is applied in the direction of specific-purpose bioreactor/fermenter, microbial measurement/inspection, combination of bioreactor/fermenter, etc. /Problems such as mutual contamination of sample wells

Active Publication Date: 2019-08-02
GUANGZHOU LBP MEDICINE SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In USD413390, USD420452, USD420745, USD420746, and USD436668, CYTOCELL Corporation discloses a method for preparing a hybrid slide of a solid carrier. This technology is to fix the probes on a raised platform to achieve physical separation. The process is complex, requiring glass cutting to create raised hillocks / platforms, which are then prepared by grinding, cleaning, polishing, etc., which is costly and must be customized for corresponding molds, and chip products for different applications cannot be prepared according to testing requirements
This method requires high requirements for cleaning and polishing after grinding, which limits the application prospects of the product
[0005] Kanglu Company proposed a method of integrating multiple probes to realize FISH detection on a glass slide. Its preparation process is discussed in detail in CN201711095572.2 and CN CN201721483366.4. The multi-probe hybridization slide It consists of probe-attached glass slides, various probes, hybridization buffer and sample-attached slides; the patent realizes the detection of bone marrow lymphocytes by preset probes (fluorescence group labeling, fluorescence reading after microscope excitation) ;The use scene setting is relatively simple; the fully hydrophobic coating design can play a certain role in separation, but when the amount of liquid added exceeds the preset 2μL, there may be overflow, and there is a risk of mutual contamination between the probe / sample well
[0006] It can be seen from the above that the existing multi-probe chips with fixed carriers still have technical deficiencies such as high production process requirements, expensive preparation, and mutual contamination in detection. Based on this, the present invention is proposed

Method used

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  • Solid phase carrier chip system for in-situ hybridization and application thereof
  • Solid phase carrier chip system for in-situ hybridization and application thereof
  • Solid phase carrier chip system for in-situ hybridization and application thereof

Examples

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Effect test

Embodiment 1

[0031] A solid-phase carrier chip system for in situ hybridization, the structure of which is as attached to the present invention figure 1 As shown, it includes eight sets of double spacers 1 and probe attachment regions 2, wherein the probe attachment region 1 is a circular hole, and the double spacer 2 is a concave-convex structure formed by a combination of straight lines and circles. , the thickness of the concave-convex structure is 20-50 μm.

[0032] The preparation method of the above-mentioned solid-phase carrier chip is: use screen printing technology to print a combination of straight lines and circles to form a concave-convex structure, the printing thickness is 20-50 μm, use the prepared double-stranded probe to pre-spot in the hole circle, and dry After drying in an oven at 45°C, the probe attachment area is formed.

[0033] The solid-phase carrier chip system described above is used for cell samples with simple morphology or less cytoplasm, such as bone marrow ...

Embodiment 2

[0036] A solid-phase carrier chip system for in-situ hybridization, which includes eight sets of double spacers and probe attachment regions, wherein the probe attachment regions are circular circles, and the double spacers are straight lines and The concavo-convex structure formed by the combination of loops and wires, the thickness of the concavo-convex structure is 20-50 μm.

[0037] The solid-phase carrier chip system also includes a sample slide, and the front side of the sample slide includes a double spacer and a sample attachment area corresponding to the solid-phase carrier chip, and the double spacer is printed by silk screen technology. The concave-convex structure is a combination of straight lines and circle lines, with a printing thickness of 20-50 μm; the formed concave-convex structure can act as a physical barrier in the subsequent hybridization process to prevent contamination between samples; the sample attachment area is a circular hole ring, and the sample ...

Embodiment 3

[0041] A method for hybridizing a cell sample using Embodiment 1 or Embodiment 2, comprising the steps of:

[0042] For probe quantification, dilute to 20ng-60ng / μL with phosphate buffer, take 1μL according to the pore size and spot on the chip carrier. Dry naturally at room temperature (humidity below 65%) or below 56 degrees.

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Abstract

The invention belongs to the field of molecular detection consumables, and relates to a solid phase carrier chip system for in-situ hybridization and application thereof. The invention provides a solid phase carrier chip system for in-situ hybridization, consisting of a probe chip, a hybridization buffer and a sample slide; the solid phase carrier chip system of the invention has the advantage oflow preparation cost, is suitable for most exfoliated cells, such as urine exfoliated cells, cervical exfoliated cells, amniotic fluid exfoliated cells, and the like, except peripheral blood lymphocyte, has two structural compositions according to the characteristics of samples, increases the physical separation between the slide and an aperture to realize a better anti-pollution effect, and realizes simultaneous detection of multiple sets of probes on a single slide.

Description

technical field [0001] The invention belongs to the field of molecular detection consumables, and relates to a solid-phase carrier chip system for in-situ hybridization and its application. Background technique [0002] In situ hybridization technology is mainly used in abnormal detection of gene fragments, including amplification, deletion, rearrangement, fusion, etc. In situ hybridization is based on a sample, a sample carrier (usually a glass slide), combined with various probes to achieve abnormality detection. Usually, one sample corresponds to one slide, and one probe is used to complete the detection. With the rapid development of genomics, this technology and product form gradually show disadvantages: First, the patient samples, sample slides, and applied detection probes detected in traditional in situ hybridization technology are separated, not a complete Product system, there is a problem of poor convenience in clinical application; second, with the promotion of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12Q1/6837C12Q1/6841
CPCC12Q1/6837C12Q1/6841C12Q2543/10C12Q2563/107C12Q2565/501
Inventor 何瑰陈绍宇
Owner GUANGZHOU LBP MEDICINE SCI & TECH