Different
types of fungal staining technique available.
1.
Lactophenol cotton blue (LPCB).
2. PAS
(Periodic Acid Schiff) reaction.
3. PAS
and GMS (Periodic Acid Schiff and Gomari methenamine Silver) reaction.
Majorly
used in Lactophenol cotton blue test
1.
Lacto phenol cotton blue test (LPCB)
staining Procedure.
·
Take
the lacto phenol cotton blue kit.
·
Lacto
phenol cotton blue solution is a mounting medium and staining agent.
·
Check
the composition.
Ingredients :
Phenol crystals------------ 20.0gm
Cotton blue----------------- 0.050gm
Lactic acid------------------ 20.0ml
Glycerol--------------------- 20.0ml
Distilled water------------- 20.0ml
Directions:
1. Take a clean glass slide.
2. add
a drop of Lacto phenol Cotton Blue reagent on a clean and dry slide.
3. Sterilize the needle and cool it.
4. Then
transfer a mycellial mat on fluid and press it
gently so that it easily mix with the stain.
5. Carefully
tease the fungal culture into a thin preparation.
6. Place a coverslip on the preparation (mycellial mat). Wait for about 5 minutes.
7. Take a blotting paper (tissue paper) and
wipe the excess stain.
8. Observe first under microscope with low power for screening in
low intensity.
Principle And
Interpretation
·
Lactophenol Cotton Blue reagent is used for
staining as well as for wet mounting of fungi.
·
Lactic
acid preserves the fungal structure and clears the tissue.
·
phenol acts as a disinfectant or fungicidal (kill
any live organisms).
·
Cotton
blue it stains the chitin in the fungal cell walls (fungal
elements).
Microscopic Examination
·
Fungal
Spores and hyphae are observed under microscope using high power (40X)
objective lens after staining with Lactophenol cotton blue.
Results
·
Fungal
spores and hyphae : pale to dark blue.
