URINE AFB :-
Procedure:-
1. Centrifuge the urine sample at 4000rpm for 8-10 minutes .
2 . Discard the supernatant and make smear with the sediment. Heat fix the smear .
3. Cover the smear with Carbol fuchsin then , apply he at till fume appears in the stain. Leave it for 5 minutes.
4. Wash the slide in tape water.
5. Decolourise the smear with 20% H2SO4 for 2-3 minutes.
6. Wash the smear in running tape water .
7. Apply methylene blue for 2-3 minutes on the smear .
8. Wash the slide in running tape water. Allow it to air dried.
9. Examine under oil immersion.
Interpretation and result :- same as sputum
TISSUE AFB STAIN:-
procedure:-
1. the tissue sections are deparaffinized in xylene and fixed in alcohol for 5 minutes .
2. Remove the slide, allow to dry the tissue sections .
3. Carbol fuchsin + heat . Leave for 5 minutes.
4. Wash the slide in tape water.
5. Decolourise with 20% H2SO4 for 2-3 minutes.
6. Counterstain with methylene blue for 2-3 minutes.
7. Wash in tape water, allowed to air dry.
8. Examine under oil immersion.
FNAC SLIDE AFB STAINING:-
Procedure :-
Overall procedure is same as sputum ,except the counterstain (MB) should kept for 5-6 minutes .
MODIFICATION IN ZN STAINING:-
Modification in decolouriser strength>>
5%H2SO4-Leprosy bacilli-
1%H2SO4-Tissue section containing 'clubs' caused by actinomyces, mycobacterium,nocardia and spores -
0.5%H2SO4-cultures of some Nocardia spp.
Other decolouriser->>
95% alcohol,
Acid alcohol(3% HCL in 95% alcohol)
Other counter stain:-
Malachite green, picric acid
COLD METHOD-kinyun's method
Carbol fuschin (4%)phenol.
In this method heat is not applied to the stain.
Stain kept 5 mins on the smear.
Washed with tape water.
*Used for cryptosporidium oocyst in fecal smears.
Procedure:-
1. Centrifuge the urine sample at 4000rpm for 8-10 minutes .
2 . Discard the supernatant and make smear with the sediment. Heat fix the smear .
3. Cover the smear with Carbol fuchsin then , apply he at till fume appears in the stain. Leave it for 5 minutes.
4. Wash the slide in tape water.
5. Decolourise the smear with 20% H2SO4 for 2-3 minutes.
6. Wash the smear in running tape water .
7. Apply methylene blue for 2-3 minutes on the smear .
8. Wash the slide in running tape water. Allow it to air dried.
9. Examine under oil immersion.
Interpretation and result :- same as sputum
TISSUE AFB STAIN:-
procedure:-
1. the tissue sections are deparaffinized in xylene and fixed in alcohol for 5 minutes .
2. Remove the slide, allow to dry the tissue sections .
3. Carbol fuchsin + heat . Leave for 5 minutes.
4. Wash the slide in tape water.
5. Decolourise with 20% H2SO4 for 2-3 minutes.
6. Counterstain with methylene blue for 2-3 minutes.
7. Wash in tape water, allowed to air dry.
8. Examine under oil immersion.
FNAC SLIDE AFB STAINING:-
Procedure :-
Overall procedure is same as sputum ,except the counterstain (MB) should kept for 5-6 minutes .
MODIFICATION IN ZN STAINING:-
Modification in decolouriser strength>>
5%H2SO4-Leprosy bacilli-
1%H2SO4-Tissue section containing 'clubs' caused by actinomyces, mycobacterium,nocardia and spores -
0.5%H2SO4-cultures of some Nocardia spp.
Other decolouriser->>
95% alcohol,
Acid alcohol(3% HCL in 95% alcohol)
Other counter stain:-
Malachite green, picric acid
COLD METHOD-kinyun's method
Carbol fuschin (4%)phenol.
In this method heat is not applied to the stain.
Stain kept 5 mins on the smear.
Washed with tape water.
*Used for cryptosporidium oocyst in fecal smears.
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