ACID FAST STAINING

SUMMARY Microbiology Lab 11th Sept

Ravi A Patel

2 PROCEDURES FOR ACID FAST STAINING

Ziehl Neelsen’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…..

ZIEHL NEELSEN’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 bacilliover greenbackground of malachitegreen

SPECIAL STRUCTURES

FLAGELLA

FLAGELLA Stained by :- Liefson’s stain

Function :- Locomotion

Composition :- Protein Flagellin

Types :- Mono-Amphi-Lopho-Peri-Atrichous

( Acronymn- P-A-A-L-M )

CAPSULEStructure 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

STRUCT. AT THE POINTERMETACHROMATIC GRANULES

METACHROMATIC GRANULES

Stained by :- L.A.M.B stain(loeffler’s Alkaline methylene blue)

Function:- Temporarily stores metabolites(carbon, phosphates)

STUCTURE AT THE POINTER SPORES

SPORES(ENDOSPORES)

Stained by:- Dorner stain

Function:- Provides resistance to bacteria in adverse conditions

Characteristic:- Spores are dormant cellsThey can undergo

germination

THANKING TO THE ENTIRE UNIVERSE