Ziehl Neelsen Method

Ziehl Neelsen Method

Ziehl-Neelsen method:
Ziehl (6) and Neelsen (7) independently proposed acid fast stain, in 1882-1883 is commonly used today. The staining reagents are much more stable than those described by Ehrlich.
The procedure for staining is as follows. Prepare a smear and fix it by gentle heat. Flood the smear with carbol fuchsin (S19) and heat the slide from below till steam rise for 5 minutes.

Do not boil and ensure that stain does not dry out. Allow the' slide to cool for 5 minutes to prevent the breakage of slide in the subsequent prevent step. Wash well with water. Decolourize the smear till red colour no longer comes out in 20% sulphuric acid. Wash with water. Counterstain with 1% aqueous solution of malachite green or Loeffler’s methylene blue (S18) for 15-20 seconds. Wash, blot dry and examine under oil-immersion objective

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