This document summarizes an acid fast staining lab performed on September 11th. It describes two acid fast staining methods - Ziehl-Neelsen's hot method and Kinyoun's cold method. Special structures observed in acid fast positive mycobacteria included flagella, capsules, metachromatic granules, and spores. Several student groups observed acid fast positive and one group observed acid fast negative bacilli under the microscope. The functions and staining properties of these special structures are also described.

1. Acid fast stainingsummary Microbiology Lab 11th SeptRavi A Patel

2. 2 procedures for acid fast stainingZiehlNeelsen’s Method (Hot Method)------The one presented as DemoKinyoun’s Method (Cold Method) ------The one which we performed today

3. Special structures we sawFlagellaCapsuleMetachromatic granulesSpores

4. Smear on the slide made from sputum of the patient having tb

5. 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 stainingAnd 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…..

6. Ziehlneelsen’s methodIn the slide we saw the Red colored +ve Acid fast stained Bacilli although the blue colored background was not so prominent..

7. Kinyoun’s methodGroup-9 got a –ve acid fast staining bacilliGroup-2,3,7 got +ve Acid fast staining BacilliThis picture is of +ve Acid fast staining bacilliIf we look closely at the pointer there is a red bacilliover greenbackground of malachitegreen

8. Special structures

9. flagella

10. flagellaStained by :- Liefson’s stainFunction :- LocomotionComposition :- Protein FlagellinTypes :- Mono-Amphi-Lopho-Peri-Atrichous ( Acronymn- P-A-A-L-M )

11. capsuleStructure at the pointer capsule and within capsule we see mycobacterium

12. CapsuleStained by :- Hiss stainFunction :- Antiphagocytosis Protects the bacteria(Virulence of the bacteria)Charateristic:- Antigenic

13. Struct. At the pointerMetachromatic granules

14. Metachromatic granulesStained by :- L.A.M.B stain (loeffler’s Alkaline methylene blue)Function:- Temporarily stores metabolites(carbon, phosphates)

15. Stucture at the pointer spores

16. Spores(endospores)Stained by:- Dorner stainFunction:- Provides resistance to bacteria in adverse conditionsCharacteristic:- Spores are dormant cells They can undergo germination

17. Thanking to the entire universe