Acid fast stainingsummary Microbiology Lab 11th Sept Ravi A Patel
2 procedures for acid fast staining ZiehlNeelsen’s Method (Hot Method) ------The one presented as Demo Kinyoun’s Method (Cold Method) ------The one which we performed today
Special structures we saw Flagella Capsule Metachromatic granules Spores
Smear on the slide made from sputum of the patient having tb
ZIEHL NEELSEN’S METHOD-hot method +VE & -VE ACID FAST STAINING------? What does it mean ------If the Organism does not take the counter stain color(In this case Methylene blue) and sustains its primary stain color(Red) then it’s a +ve acid fast staining And Vice Versa…………..for –ve Acid Fast staining * Involves heating for 5 min on slow burner after being stained with primary stain so called Hot Method unlike the Kinyoun’s in which we did not use burner after primary staining…..
Ziehlneelsen’s method In the slide we saw the Red colored +ve Acid fast stained Bacilli although the blue colored background was not so prominent..
Kinyoun’s method Group-9 got a –ve acid fast staining bacilli Group-2,3,7 got +ve Acid fast staining Bacilli This picture is of +ve Acid fast staining bacilli If we look closely at the pointer there is a red bacilli over green background of malachite green
flagella Stained by :- Liefson’s stain Function :- Locomotion Composition :- Protein Flagellin Types :- Mono-Amphi-Lopho-Peri-Atrichous ( Acronymn- P-A-A-L-M )
capsule Structure at the pointer capsule and within capsule we see mycobacterium
Capsule Stained by :- Hiss stain Function :- Antiphagocytosis Protects the bacteria(Virulence of the bacteria) Charateristic:- Antigenic