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Ex. 5: Acid-Fast and Endospore Stains
Objectives:Explain the rationale and procedure for the acid-fast and endospore stains
Perform and interpret the acid fast and endospore stains
Review the structure and function of endospores
For Acid Fast Bacteria
Use only
Carbol fuchsin for AFB!!
Acid Alcohol
MethyleneBlue
Modified Kinyoun’s Acid-fast Stain – Cold Method
• Heat fix smear
• Primary stain: Carbol fuchsin for AFB, (5 min)
• Rinse with water
• Decolorize briefly (~10 drops) with acid alcohol (~ 30 sec)
• Immediately rinse with water
• Counterstain: Methylene blue (1 min)
• Rinse with water
Endospore Stain
• Heat fix smear
• Primary stain: Malachite green, steam (5 min)
• Rinse thoroughly with water (~ 30 sec)
• Counterstain: Safranin (1 min)