Acid resistant staining method

A technology of acid-fast dyeing and dyeing agent, which is applied in the field of acid-fast dyeing, can solve the problems of different uses and practical significance, and the reference value of antibacterial dyeing method is not great, so as to facilitate commercial transportation, dyeing standardization and Effects of automation and logistics cost reduction

Inactive Publication Date: 2015-04-29
SUZHOU WEILITE BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Gram staining and acid-fast staining are two different staining methods in bacteriology, and their uses an

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Take clean glass slides and materials to be tested (body fluids) to smear according to routine requirements, dry and fix with flame or acetone. First add 3ml of primary staining agent (distilled aqueous solution containing 1.6wt% basic fuchsin and 6.4wt% phenol) dropwise to cover the specimen, stain for 2 minutes, remove the dye solution obliquely without washing, then add 3ml dropwise for decolorization and complexation Dye (0.8wt% methylene blue solution) covers the specimen, stains for 30 seconds, washes with water, and examines under the microscope after drying. It can be seen in the smear that the acid-positive bacteria stained red and the acid-negative bacteria stained blue are in sharp contrast.

Embodiment 2

[0037] Take clean slides and materials to be tested (secretion submitted for inspection) to smear according to routine requirements, dry and fix with flame or acetone. First add 2ml of primary staining agent (an acetone solution containing 1.0wt% basic fuchsin and 5.4wt% phenol) dropwise to cover the specimen, stain for 1 minute, remove the dye solution obliquely without washing, then add 2ml dropwise for decolorization and recombination Dye (0.8wt% methylene blue solution) covers the specimen, stains for 30 seconds, washes with water, and examines under the microscope after drying. In the smear, acid-positive bacteria stained red and acid-negative bacteria stained blue contrast sharply.

Embodiment 3

[0039] Take clean slides and materials to be tested (bacterial culture) to smear according to routine requirements, dry and fix with flame or acetone. First add 1ml of primary staining agent (an ethanol solution containing 2.0wt% basic fuchsin and 7.0wt% phenol) dropwise to cover the specimen, stain for 1.5 minutes, remove the dye solution obliquely without washing, then add 1ml dropwise for decolorization and complexation Dye (0.8wt% methylene blue solution) covers the specimen, stains for 30 seconds, washes with water, and examines under the microscope after drying. It can be seen in the smear that the acid-positive bacteria stained red and the acid-negative bacteria stained blue are in sharp contrast.

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PUM

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Abstract

The invention relates to an acid resistant staining method. The method is characterized by comprising the following steps: (1), putting a material smear to be detected on a slide, and drying and fixing the material smear to be detected so as to obtain a sample; (2), dripping primary stain to cover the sample, and after staining for 1-2 minutes, removing the primary stain; (3), dripping decolorizing counterstain to cover the sample, after staining for 25-35 seconds, washing the sample by water, and performing microscopic examination after drying, wherein the primary stain is a solution comprising 1.0-2.0 weight percent of basic fuchsin and 5-7 weight percent of phenol, and the decolorizing counterstain is a methylene blue solution of 0.6-0.8 weight percent. Compared with the prior art, the method disclosed by the invention has the advantages that heating is selectable in staining; the method can be used for manual and automatic staining, and is beneficial to the standardization and the automation of staining; the method disclosed by the invention is reliable, and the result is distinct and is easy to observe and judge.

Description

technical field [0001] The invention relates to a bacterial staining method, in particular to an acid-fast staining method. Background technique [0002] Acid-fast staining method is a bacterial staining method initiated by F. Ehrlich in 1882 and improved by F. Ziehl. The most representative ones are Ziehl-Neelsen staining and Ziehl-Gabbet staining for Mycobacterium tuberculosis. After staining with carbofuran, decolorize with hydrochloric acid ethanol, and contrast staining with methylene blue. [0003] The principle of acid-fast staining is as follows: the cell wall of mycobacteria (including Mycobacterium tuberculosis) contains a large amount of lipids, surrounded by peptidoglycan, so mycobacteria are generally not easy to stain, and it is necessary to heat and prolong the staining time to promote its staining. coloring. However, after the mycolic acid in mycobacteria is combined with the dye, it is difficult to be decolorized by the acid decolorizing agent, so it is c...

Claims

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Application Information

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IPC IPC(8): G01N1/30
Inventor 叶鹰叶鸿杨玄墨徐燕萍徐晨
Owner SUZHOU WEILITE BIOLOGICAL TECH CO LTD
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