A method for staining apoptotic cells

A staining method and technology of apoptotic cells, which are applied in the preparation, sampling, and instrumentation of test samples, can solve the problems of high requirements for the staining process, expensive reagents, and damage to the tissue structure of slices, and achieve low dyeing costs and cheap reagents Easy-to-obtain, simple-to-operate effects

CN107782598BActive Publication Date: 2020-09-29SICHUAN ACAD OF CHINESE MEDICINE SCI
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2020-09-29

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Abstract

The invention provides an apoptotic cell staining method. The apoptotic cell staining method includes the steps of (1), dewaxing sections to water and staining for 0.2-5 minutes by a staining solutionI, wherein the staining solution I comprises 0.1-1 g of acid fuchsin, 0.1-2 g of ponceau, 0.1-3 g of orange G and 300ml of 0.2% acetic acid; (2), washing by an acetic acid solution, treating for 3-8minutes by a phosphomolybdic acid solution, and washing by the acetic acid solution again; (3), staining for 10-30 minutes by a staining solution II and washing by the acetic acid solution again, wherein the staining solution II comprises 0.1-1 g of brilliant green and 100 ml of 0.2% acetic acid; (4), performing dehydration and transparency treatment prior to mounting. The apoptotic cell stainingmethod can identify apoptotic cells at different phases and displaying cell apoptosis statuses comprehensively, and is simple to operate, low in staining cost and suitable for popularization and application.
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Description

Technical field

[0001] The invention belongs to the field of cell identification, and specifically relates to a staining method for apoptotic cells. Background technique

[0002] Apoptosis is the death of a single cell or a small group of cells in a living body. The plasma membrane of the dead cell is not broken, it does not cause the autolysis of the dead cell, and it does not cause an acute inflammatory reaction. The occurrence of apoptosis is related to gene regulation, and it is also called programmed cell death. Apoptosis is a basic physiological mechanism of the body, and it runs through the entire life activities of the body, including the development of embryos, the alternation of old and new cells, the normal degeneration and atrophy of certain tissues, or the spontaneous inhibition of tumor cells.

[0003] At present, the conventional in situ identification technology of apoptotic cells is mainly a staining method in addition to observation by electron microscope. [0004...

Examples

Embodiment 1

[0034] The staining object is a serial section of liver injury model after giving mice concanavalin, the staining result is as follows figure 1 As shown, using TUNEL in situ hybridization staining method as a control experiment, the control staining results are as follows figure 2 Shown. In the figure, although red blood cells are also marked with the same color as apoptotic cells, the morphology of red blood cells is significantly different from other tissue cells, and there is basically no interference in the recognition of apoptotic cells.

[0035] Follow the steps below to dye:

[0036] (1) Dewax the slices to xylene for 40 minutes, treat with absolute ethanol for 10 minutes, and then treat with 95% ethanol, 90% ethanol, 80% ethanol, 70% ethanol, 60% ethanol, 50% ethanol for 2 minutes each In, after washing with water, dye with dyeing solution I for 5 minutes;

[0037] (2) After cleaning with acetic acid aqueous solution, treat with phosphomolybdic acid aqueous solution for 8 m...

Embodiment 2

[0050] Except that the staining object is a serial section of tumor tissue grown from h22 tumor cells planted in the armpit of a mouse, the rest are the same as in Example 1. The staining results are as follows image 3 Shown. The TUNEL in situ hybridization staining method is also used as a control experiment, and the results of the control staining are as follows Figure 4 Shown.

Embodiment 3

[0052] Except for the formula of dyeing solution I:

[0053] Except for 0.1 g of acid complex red, 0.1 g of ponceau red, 0.1 g of orange G, and 300 ml of 0.2% acetic acid, the rest are the same as in Example 1.

[0054] The staining results are consistent with Example 1, which can distinguish apoptotic cells.