Microbiology lab.

Report

Name: Acid fast stain Ziehl-Neelsen method.

Aim: To identify if the bacteria is resist to decolorization by acid daring

laboratory staining procedure.

Material: smear, glass slide, tap water, carbolfuchin stain (filtered), 20%
of 𝐻2 𝑆𝑂4 or mix of acid alcohol, counter stain, methylene blue 5g/l,
methylene green 5g/l (0.5% w/v), Bunsen burner.

Procedure: 1. Make a smear on the glass slide and allow to heat-fix.

2. Flood the carbolfuchsin on the slide and & warm under the spirit lamp
for 5 min until vapor.
3. Allow to cool and wash under tap water.
4. by 20% solution of 𝐻2 𝑆𝑂4 or a mix of acid alcohol (3%HCL in 95%
Ethanol)slowly drop wise until the dye no longer runs off from the smear
for (10- 30) sec.
5. Rinse with water.
6. Counter stain with methylene blue for 2 minute.
7. Wash under tap water and allow drying.

Result:
Reagent Acid fast Non-acid fast
Carbol fusion with heat Red (hot pink) Red (hot pink)
Acid alcohol Red Colorless
Methylene Blue/ Red Blue/Green
Malachite Green

Conclusion: The acid fast bacteria resist the decolorization by acid, while
the non-acid fast bacteria doesn't resist the decolorization by acid during
the stain procedure.

Miami arif
Group B3