3.10: Acid-Fast Stain
- Page ID
- 90987
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Preparation
- Have test tubes with sterile TSB on hand.
- 12-18 hours before the laboratory, inoculate test tubes of sterile TSB with a mixture of Staphylococcus epidermidis and Mycobacterium chelonae and incubate at 37 °C .
Materials
- 12-18 hour cultures (enough for groups to share)
- plain microscope slides (at least 1 per group)
- Bunsen burners (1 per group)
- inoculating loops (1 per group)
- (if using bacteria on solid medium rather than TSB) droppers with saline, DI water, or distilled water
- slide warmer
- test tube racks (1 per group plus one or more racks to hold cultures)
- 400-500 mL beakers with ~200 mL of water each (1 per group)
- hot plate (1 per group)
- slide rack hat fits over beaker (1 per group)
- paper towels
- carbol fuchsin in dropper bottles (1 per group)
- acid-alcohol in dropper bottles (not ethanol from the Gram stain kit) (1 per group)
- methylene blue in dropper bottles (1 per group)
- wood test tube clamps/holders (1 per group)
- small chemical waste containers (1 per group)
- distilled or DI water in squirt bottles (1 per group)
- bibulous paper (enough for each groups to blot their slides)
- light microscopes (1 per group)
- colored pencils
- (optional, but recommended) Wax pencils (1 per group)
- (if using oil immersion) immersion oil dropper bottles
- (if using oil immersion) lens paper