Acid Fast Staining.

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Presentation transcript:

Acid Fast Staining

Kinyoun Acid-Fast Staining Procedure Carbol fuchsin Acid alcohol Methylene blue Reddish-pink Blue Acid Fast Nonacid Fast Kinyoun Acid-Fast Staining Procedure 1. A sample of cells is mixed with a drop of water on a clean slide to make a smear. After air drying, the slide is heat fixed. 2. Slide is flooded with carbol fuchsin (primary stain basic fuchsin + mordant carbolic acid) and allowed to sit for 15 minutes. Slide is rinsed until water coming off the slide is clear. 3. Slide is decolorized with acid alcohol (3% HCl and 95% alcohol – ACID ALCOHOL) 20 seconds, then rinsed. 4. Slide is flooded with methylene blue (counter stain) for 60 seconds and then rinsed.

Note the reddish-pink color and “cording” (sticking together in long ropy masses) of the Mycobacterium cells due to the excess lipids of the cell wall

Identify the following in this sputum smear: Epithelial cell (E), Streptococcus mutans (S), Mycobacterium tuberculosis (M) M S E

Locate Mycobacterium tuberculosis (M)

Put these solutions in order of their use in the Kinyoun acid-fast staining procedure B C