Antacid staining test of sputum specimens from patients with pulmonary tuberculosis

Summary

Mastery of antacid staining of sputum specimens.

Operation method

antacid staining

Principle

Mycobacterium is generally not easy to color, after heating or prolonging the staining time and coloring, it can resist the decolorizing effect of acid and alcohol, so it is also known as acid-resistant bacilli. Mycobacteria that are pathogenic to humans are mainly Mycobacterium tuberculosis and Mycobacterium leprae.

Move

1. Take a smear (slightly thicker) of the thick purulent or caseous blood-streaked portion of the sputum, dry it naturally, fix it through a flame, and make a circle around the periphery of the smear with a wax pencil.

2. Hold the slide with a wooden clamp, add drops of carbolic acid red dye to cover the surface of the specimen, and increase the temperature until it vapors on the high side of the flame and maintain it for 5 minutes (be careful not to allow the dye to boil or boil dry), and add more dye to replenish the dye when the dye evaporates to a lesser extent.

3. Allow the specimen to cool and then wash.

4. Decolorize with 3% hydrochloric acid alcohol until no red dye runs down.

5. After washing, re-stain with Lüchner's blue for 1 min, wash with water, absorb and examine microscopically. Acid-resistant bacilli showed red color, background and non-acid-resistant bacilli showed blue color.

Common Problems

1. What are the similarities and differences in the morphological staining of Mycobacterium tuberculosis and Mycobacterium leprae?

2. What are the culture characteristics of Mycobacterium tuberculosis?

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