Improved frozen slicing method and applications thereof

A frozen section and slicing technology, which is applied in the field of tissue sample preparation and pathological research, can solve problems such as the inability to completely maintain the fine structure of the tissue, and achieve the effects of short time, high quality and low cost.

Inactive Publication Date: 2015-05-20
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But commercial kits are not yet able to fully preserve the fine structure of the tissue

Method used

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  • Improved frozen slicing method and applications thereof
  • Improved frozen slicing method and applications thereof
  • Improved frozen slicing method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Application of the Improved Frozen Section Method in Example 1 in Mouse Embryo Samples

[0069] The embryo sample is a tissue sample with a large water content and is very delicate. It is very difficult to obtain a good tissue fine structure at the same time by freezing the whole embryo. The application of the method of the present invention successfully obtains continuous sections of the complete embryo, and the tissue structure remains intact. High-quality RNA, the specific experimental operation is as follows:

[0070] 1. Frozen section processing steps of experimental materials

[0071] 1) Take mouse embryos on day E15 (the time when the vaginal suppository is observed after the male and female mice are caged together is defined as day E0), measure the head and hip length of 15mm, wash them in PBS solution, and then wash them off with dust-free absorbent paper Surface moisture, then soak in 4ml 20% DMSO solution (solvent: 20v / v% ethanol aqueous solution) at 4°C for...

Embodiment 2

[0080] Application of the improved frozen section method of embodiment 2 in mouse kidney tissue

[0081] The method of the present invention can not only be applied to embryonic samples with very high water content, but also to other tissues with high water content, such as the kidney of mice. In this embodiment, the method of the present invention is applied to the kidney tissue of mice, and many Good results, the specific experimental method is as follows.

[0082] 1. Frozen section processing steps of experimental materials

[0083] 1) Take the kidney tissue of the mouse, wash it in PBS solution, absorb the surface moisture with dust-free absorbent paper, then put it into 4ml of 10% DMSO aqueous solution at 25°C for 30 minutes, take it out, and absorb it with dust-free absorbent paper Remove the surface liquid, and then soak in OCT embedding medium at 25°C for 2 hours.

[0084] 2) The mouse kidney tissue treated in step 1) was embedded with a new OCT embedding agent, and ...

Embodiment 3

[0091] Example 3 Application of Improved Frozen Section Method in Adult Mouse Brain Tissue

[0092] In addition to being applicable to tissues with very high water content (such as embryos and kidneys), the method of the present invention is also applicable to tissues with high lipid content that are difficult to handle, such as the brain of mice. This example is based on the brain tissue of mice Apply the method of the present invention, also obtained good result, concrete implementation method is as follows.

[0093] 1. Frozen section processing steps of experimental materials

[0094] 1) Take the brain tissue (15mm×10mm) of an adult mouse of the C57BL / 6J strain, wash it in PBS solution, absorb the surface moisture with a dust-free absorbent paper, and then put it into 4ml of 20% DMSO solution (the solvent is HBSS buffer solution ) at 0°C for 1.5h, take it out, absorb the surface liquid with dust-free absorbent paper, and then put it into OCT embedding medium and soak at 0°...

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Abstract

The invention relates to the field of tissue sample preparation and pathogenic research, and particularly relates to an improved frozen slicing method and applications thereof. The improved frozen slicing method comprises the following steps: tissue pretreatment step, tissue embedding step, tissue freezing and sliding steps, wherein frozen tissue slices can be obtained after the slicing step, and a freezing protecting agent is adopted for soaking a tissue sample in the tissue pretreatment step. According to a tissue sample preparation method, not only can a fine tissue structure similar to a wax slice effect be obtained, but also the high quality of biological macromolecules, such as DNA, RNA and protein, can be guaranteed so as to meet the whole genome detection and analysis of the biological macromolecules. The improved frozen slicing method has the advantages of being simple, strong in universality, short in sample preparation time, wide in application prospect, and the like.

Description

technical field [0001] The invention relates to the field of tissue sample preparation and pathological research, in particular to an improved frozen section method for preparing tissue samples and its application. Background technique [0002] With the gradual completion of genome sequencing of humans and other species, modern biological research has entered the post-genome era—the functional genome era. With the development of functional genomes, high-throughput technologies for the study of different biomacromolecules have been developed, including genome technology, transcriptome technology, proteome technology, etc. In recent years, various omics methods for small or even single-cell samples have also begun to be developed and applied. With the need for massive data processing and analysis, bioinformatics is also developing rapidly. At the same time, in the sorting of tissue samples, with the application of laser microdissection technology, it is possible to sort spec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 郭妍张小丹邵志峰高维强狄文王酉李青
Owner SHANGHAI JIAO TONG UNIV
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