Tissue permeabilization method for spatial transcriptome sequencing

A technology of spatial transcription and organization, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of high price, and achieve the effect of low cost, good repeatability and strong operability

Pending Publication Date: 2022-05-27
BIOMARKER TECH
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  • Description
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Problems solved by technology

However, the commonality of the above three platforms is that they are expensive. Among them, the widely used 10x Visium has an average market price of 35,000 yuan per sample.

Method used

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  • Tissue permeabilization method for spatial transcriptome sequencing
  • Tissue permeabilization method for spatial transcriptome sequencing
  • Tissue permeabilization method for spatial transcriptome sequencing

Examples

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Embodiment 1

[0071] The following examples are intended to illustrate the present invention, but not to limit the scope of the present invention. Unless otherwise specified, the examples are in accordance with conventional experimental conditions, such as Sambrook et al. Molecular Cloning Laboratory Manual (Sambrook J&Russell DW, Molecular Cloning: a Laboratory Manual, 2001), or in accordance with the conditions suggested by the manufacturer's instructions. Example 1 Spatial transcriptome tissue permeabilization optimization method

[0072] This embodiment provides a method for optimizing spatial transcriptome tissue permeabilization. For the specific process, see Figure 8 .

[0073] It includes the following steps:

[0074] 1. Tissue patching, fixation and staining

[0075] 1.1 Slice the fresh frozen tissue (rat embryo sample) and stick it on the permeabilization chip, then put the permeabilization chip with the attached tissue into 40ml of pre-cooled methanol, and fix it at -20°C for...

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Abstract

The invention provides a tissue permeabilization method for space transcriptome sequencing. The tissue permeabilization method comprises the steps of tissue pasting, fixing and dyeing, tissue permeabilization, fluorescent cDNA synthesis, tissue removal, fluorescent scanning imaging and the like. By using the method, the tissue sample can be subjected to permeabilization, reverse transcription and tissue removal, and the optimal permeabilization condition is selected according to the fluorescence signal. Compared with the traditional method, the method has the advantages of wide compatible range, strong operability, good repeatability, low cost and the like. The method is mainly applied to the tissue optimization process of the space transcriptome.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a tissue permeabilization method for spatial transcriptome sequencing. Background technique [0002] In multicellular organisms, the gene expression of a single cell occurs strictly in a specific temporal and spatial order, that is, gene expression is temporally and spatially specific. Time specificity can be obtained by taking samples at different time points and using single-cell transcriptome sequencing technology to resolve cell types and gene expression patterns in the time dimension. Spatially specific information is relatively difficult to obtain. Both conventional transcriptome sequencing and single-cell transcriptome sequencing are difficult to restore the original location information of cells. Traditional in situ hybridization technology is difficult to achieve high-throughput detection. Therefore, it is particularly necessary to develop spatial transcriptome te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2527/125C12Q2521/537C12Q2521/107C12Q2527/127C12Q2565/519C12Q2563/107
Inventor 郑洪坤刘敏张梦龙刘晨阳
Owner BIOMARKER TECH
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