WADE FITE TECHNIQUE

                                      WADE FITE TECHNIQUE


Principle  These organisms posses a capsule containing long chain fatty acid, (mycolic
acid), which influences the penetration and resistance to removal of stain by acid and alcohol. The avoidance of defatting agents such as alcohol and Xvlene in the
method are an attempt to conserve its fragile fatty capsule.



Solution


Carbol fuchsin ( Pink color)
Strong Basic fuchsin                         -           1 gm
Absolute Alcohol                               -           10 ml
5%aqueous phenol                           -           100 ml
Filter before use.


Methylene blue

Methylene blue                      -             l gm
Distilled water                                     -           100ml


Peanut Oil- Xylene mixture

Peanut oil                    -           1 Part
Xylene                         -             2 Part




Procedure


1.    Keep the section in peanut oil-Xylene mixture for 30 mnts
2.    Drain the excess oil, blot it with filter paper.
3.    Wash it in water.
4.    Stain it with strong carbol fiuchsin for  30mnts. (RT)
5.    Wash it in water
6.    Differentiate with 1% acid alcohol, until the section becomes colourless.
7.    Keep it in running tap water for 5-10 mnts.
8.    Counterstain with methylene blue for 30 secs.
9.    Wash it in Water, dehydrate, clear and mount.


Result

 Mycobacterium Laprae         -        Pink
Background                                       -           Blue


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