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Space transcriptome detection chip and method

A detection chip and spatial transcription technology, applied in biochemical equipment and methods, bioreactor/fermenter combination, biochemical instruments, etc., can solve the problems of limited number of transcripts, lack of spatial information of tissue cells, etc., and achieve high resolution Effect

Inactive Publication Date: 2019-05-17
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Currently, tissue transcriptomes are typically studied by RNA-sequencing of homogeneous biopsies, resulting in loss of tissue-cell spatial information
Or methods exist for cellular location monitoring, but they are limited in the number of transcripts that can be analyzed and rely on large existing datasets, expensive instrumentation, or extensive effort

Method used

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  • Space transcriptome detection chip and method

Examples

Experimental program
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Effect test

Embodiment 1

[0038] 1. Preparation of microplates

[0039] Design the size of the microwell plate 3 according to the experimental scale, the size of the well plate is 3 mm×3 mm, and microholes are etched on the silicon wafer as the initial mold, the depth of the microwells is 20 μm, the diameter of the microwells is 15 μm, and the spacing between the holes is 4 is 10 μm.

[0040] Pouring polydimethylsiloxane (PDMS) on the silicon wafer, and taking off the PDMS after molding, a reverse mold with micropillars can be made, and agarose prepared with enzyme-free water at a concentration of 5%, heated After melting, pouring is condensed on the reverse mold (PDMS microcolumn plate), and the agarose plate is peeled off to form a microporous plate 3 with a certain thickness. Add the DPBS-EDTA mixture that is harmless to the cells on the surface of the microwell plate 3 during storage, cover and store in a refrigerator at 4°C, and then the microwell plate 3 can be prepared, which can ensure that th...

Embodiment 2

[0065] Adult mice were euthanized and their brain tissues were taken, and tissue slices were prepared according to the methods in the examples.

[0066] All the other steps are the same as in Example 1.

[0067] After the construction of the gene sequencing library, the paired sequencing was performed on the Illumina NextSeq platform, and the gene expression profile was obtained after splitting, screening and comparison.

Embodiment 3

[0069] Adult mice were euthanized and their liver tissues were collected, and tissue sections were prepared according to the method in the examples.

[0070] All the other steps are the same as in Example 1.

[0071] After the construction of the gene sequencing library, the paired sequencing was performed on the Illumina NextSeq platform, and the gene expression profile was obtained after splitting, screening and comparison.

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Abstract

The invention provides a space transcriptome sequencing chip and method. The chip and the method are used for determining one or more single cell sequences in different space positions in tissues. Thechip comprises a micropore plate and coding sequencing microspheres, wherein the micropore plate is provided with the coding sequencing microspheres. The method comprises the steps of coupling with the microspheres using a plurality of random codes, wherein each random code comprises a molecular marker and a position marker barcode; the molecular marker can detect sequence information of single cells of the tissues, and the position markers are used for recording space position information of the single cells of the tissues. Based on the detection chip and the detection method, the single-cell specific transcriptome information of the tissues can be analyzed and detected, and the spatial positions of thousands of single cells in the tissues can be simultaneously detected.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a space transcriptome sequencing chip and method. Background technique [0002] Traditional transcriptome research, in most cases, is carried out on a large number of mixed cells, and it is impossible to observe subtle differences between individual cells, covering up the behavior of independent individual samples and the specificity and cell differences in life phenomena. The research on a single cell is the limit state pursued by cell life analysis technology and the ultimate challenge to the development of traditional technologies. Multicellular masses such as tissues and tumors can include heterogeneous cellular environments. These complex cellular environments can often exhibit multiple phenotypes that can be indicative of multiple genotypes. Distilling multicellular complexity to single-cell variability is an important aspect of understanding multicellular heterogeneity. [...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12M1/00
Inventor 赵祥伟郭云霞涂景葛芹玉
Owner SOUTHEAST UNIV
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