THE VIRTUAL EDGE: Lab 3 Bacterial Staining Techniques II |
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Acidfast Stain: Background and IntroductionMycobacterium and many Nocardia species are called acid-fast because during an acid-fast staining procedure they retain the primary dye carbol fuchsin despite decolorization with the powerful solvent acid-alcohol. Nearly all other genera of bacteria are nonacid-fast. The acid-fast genera have lipoidal mycolic acid in their cell walls. It is assumed that mycolic acid prevents acid-alcohol from decolorizing protoplasm. The acid-fast stain is a differential stain. Ziehl Neelsen Acid-fast stain
Step 2: Smear Preparation (Review)
Step 3:Cover the smear with carbolfuchsin dye. Carbolfuchsin a potential carcinogen. Please wear gloves and work with the stain in the hood. Place a piece of paper towel on top of the dye. Be sure the paper towel is saturated with the dye. Step 4:Dry heat for 2 minutes. Step 5:Cool and rinse with water. Decolorize with acid-alcohol for 15-20 seconds. Step 6:Wash the top and bottom of slide with water and clean the slide bottom well. Step 7:Counterstain with Methylene Blue for 30 seconds to 1 minute. Wash and blot the slide with bibulous paper. Focus 10X - then use oil immersion. (Oil immersion review)
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Lab 3 / Gram Stain / Acid Fast Stain / Lab 3 Organisms Please take a few minutes to fill out a brief survey about your experience using the Virtual Edge: https://docs.google.com/forms/d/1yGbkF0KM92WBSk-IgS-EkjxkTKTQwhzuXmDsVpwRDoU/viewform Please email comments/problems to cboggs@uwyo.edu |
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Rachel Watson, M.S. |
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