Stain for acid-fast bacilli
AssigneeBecton, Dickinson and Company
InventorJayakumar Reuben
Life of this patent
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Abstract
Acid-fast bacilli are stained with a primary stain that does not require heating and then with a decolorizing counter-stain that does not require a separate acid or alcohol decolorization step. The primary stain includes basic fuchsin, ethyl alcohol, isopropyl alcohol, phenol and water. The counter-stain includes methylene blue, ethyl alcohol, potassium hydroxide, glycerol, glacial acetic acid, polyvinylpyrrolidone and water.
Claims4
A staining system for acid-fast, bacilli comprising a primary stain and a separate decolorizing counter-stain wherein
- said primary stain is an aqueous solution containing for each 100 ml of solution the following constituents:
- from 1.75 to 2.25 gm basic fuchsin;
- from 16 to 18.5 ml ethyl alcohol;
- from 0.925 to 1.025 ml isopropyl alcohol;
- and
- from 7.2 to 7.8 gm phenol;
- and
- said separate decolorizing counter-stain is an aqueous solution containing for each 100 ml of solution the following constituents
- from 0.2 to 0.25 gm methylene blue;
- from 20 to 25 ml ethyl alcohol;
- from 0.0075 to 0.0125 gm potassium hydroxide;
- from 9.5 to 10.5 ml glycerol
- from 4 to 5 ml acetic acid;
- and
- from 0.05 to 0.1 polyvinylpyrrolidone (0.01 V/V % solution).
A method for staining a slide in preparation for microscopic observation of acid-fast bacili comprising:
- preparing a slide with a suspected mycobacteriological sample;
- applying a primary stain to the slide wherein said primary stain is an aqueous solution containing for each 100 ml of solution the following constituents:
- from 1.75 to 2.25 gm basic fuchsin;
- from 16 to 18.5 ml ethyl alcohol;
- from 0.925 to 1.025 ml isopropyl alcohol;
- and
- from 7.2 to 7.8 gm phenol;
- incubating the slide for a first period of about to about 6 minutes;
- rinsing the slide;
- applying a decolorizing counter-stain to the slide wherein the decolorizing counter-stain is an aqueous solution having for each 100 ml of solution the following constituents:
- from 0.2 to 0.25 gm methylene blue;
- from 20 to 25 ml ethyl alcohol;
- from 0.0075 to 0.0125 gm potassium hydroxide;
- from 9.5 to 10.5 ml glycerol
- from 4 to 5 ml acetic acid;
- and
- from 0.05 to 0.1 ml polyvinylpyrrolidone 0.01 V/V % solution
- inoculating the slide for a second period of from about 2 to about 4 minutes;
- and
- rinsing the slide.
Dependencies are inferred from the claim text.