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Index >> Staining Methods in Microbiology >> Staining Method of Gross

Staining Method of Gross

Staining Method of Gross
This method is also known as cold method (8), because it eliminates the heat treatment during the basic fuchsin application. The method is rapid and reproducible. Increase in concentration of basic fuchsin and a little addition of detergent will allow the acid-fast organisms to take the stain at room temperature. The sequence of steps to be followed is as under. Prepare the smear and fix it by heat.

Stain the smear with basic fuchsin with Tween-80 solution (S27) at room temperature for 5-10 minutes. Rinse the smear with water. Decolourize the smear with 3% HCl in ethanol. Rinse the slide with water. Counterstain with Loeffler's methylene blue (S18) for 3 minutes. Wash, blot dry and examine under oil-immersion objective. Acid-fast organisms appear red, while non acid-fast organisms and background apper blue in colour.

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