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Acid Fast Staining

3rd sem notes

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6 views

Acid Fast Staining

3rd sem notes

Uploaded by

vinaybainada10000
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Acid Fast Staining

Dr. Richa Sharma


Assistant Prof
Microbiology
Acid Fast Staining or Ziehl Neelson
Staining (ZN Staining)
• This staining was first developed by Paul Ehrlich
in 1882 and modified by Ziehl Neelson so it is also
called Ziehl Neelson Staining (Z-N Staining).
• This is a differential staining because it separate
two types of bacteria.
• 1 Acid Fast Bacteria- Which give pink or red color
of Carbol fuschin.
• 2 Non Acid Fast Bacteria: Which give blue color of
methylene blue dye
Acid Fast Staining
• This method is used for those microorganisms
which are not staining by simple or Gram
staining method, particularly the member of
genus Mycobacterium, are resistant and can
only be visualized by acid-fast staining.
• Presence of long chain fatty acids or Mycolic
acid in the cell wall of tubercule bacilli.
Procedure of Acid Fast or ZN Staining
• Smear formation from the cough or Sputum
Sample

• Add Concentrated Carbol fuschin and gently


heated until the steam arises and left for 5-10
mins.
Procedure of Acid Fast or ZN Staining
WASH WITH TAP WATER

Add decolorizing agent (20% sulphuric acid) for 30


secs-1 min.

WASH WITH TAP WATER


Procedure of Acid Fast or ZN Staining
WASH WITH TAP WATER

Add Methylene blue (Counterstain) for 2 min.

WASH WITH TAP WATER

Observe under Microscope.


Observation
• Observe the slide for the identification of acid
fast bacilli and non acid fast bacilli
• Acid fast bacilli are rod shaped and red/pink in
color.
• Non Acid Fast Bacilli are appear dark blue in
color.
Result
• Acid Fast Bacili are bright red or pink in color
on a blue color background and non acid fast
will appear dark blue in color.
Acid Fast Bacilli (Red color thread like)
in a blue color background.
Precautions
• 1 Smear Should be prepared properly.
• 2 Staining should be done properly.
• 3 During smear formation from sputum
sample, always wear PPE.
THANKS

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