Cell three-dimensional culture matrigel wax embedding method

Abstract

The invention discloses a cell three-dimensional culture matrigel wax embedding method. The method includes steps: (1) designing and manufacturing a slide which morphologically fits an internal storage space of an embedding box; (2) cutting off a matrigel three-dimensional culture tissue from a Transwell upper chamber, and fixing in paraformaldehyde in mass percent of 4-6% for 6-24h; (3) after successful fixing, taking the matrigel three-dimensional culture tissue out of paraformaldehyde, and enabling a collagen face of the matrigel three-dimensional culture tissue to be adsorbed on the slide;(4) putting the slide which morphologically fits the internal storage space of the embedding box and adsorbs the matrigel three-dimensional culture tissue into the embedding box in which gauze is laid; (5) dewatering, transparentizing and waxing; (6) adopting a tissue slicing knife for completely scrape off the tissue adsorbed on the slide from the slide, and embedding the tissue. In an embeddingprocess, tissue curling and shedding and cell falling are avoided. Therefore, the method is suitable for popularization and utilization.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a gel wax embedding method for a three-dimensional cell culture substrate. Background technique [0002] Biological tissue embedding is a commonly used method in the field of biotechnology. Traditional embedding methods include paraffin embedding and plastic embedding. The former has a long production process (up to about 2 days). The lipid components in the tissue are not easy to retain, easily cause tissue cell shrinkage, have adverse effects on the activities of most enzymes, and cannot be used for source floating staining; the latter has the disadvantages of not being able to perform immunohistochemical staining and requiring expensive supporting equipment Insufficient equipment and so on. Although the cryostat microtome used in the 1970s has high speed and can not only perform enzyme histochemical staining, but also immunohistochemical staining, it is difficult to ...

AI Technical Summary

Problems solved by technology

The lipid components in the tissue are not easy to retain, easily cause tissue cell shrinkage, have adverse effects on the activities of most enzymes, and cannot be used for source floating staining; the latter has the disadvantages of not being able to perform immunohistochemical staining and requiring expensive supporting equipment lack of equipment

Although the cryostat microtome used in the 1970s has high speed and can not only perform enzyme histochemical staining, but also immunohistochemical staining, it is difficult to make thinner sections and serial sections below 5 μm, and because of Defects in which the cell structure is unclear due to freezing

[0004] 1. Matrigel is easy to curl during dehydration due to its loose structure

[0005] 2. The tissue is easy to fall off and lose in the process of dehydration, transparency and wax dipping

[0006] 3. Cells on the surface of the tissue are easy to fall, which reduces the speed and efficiency of observing and studying tissue lesions

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