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BACKGROUND
Fungus are a group of eukaryotic protists that basically lacks chlorophyll. Depending upon various species the fungus have various morphological features. The objective of this test is to become familiar with various mycological culture techniques and identification of structural components of fungus.1
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REQUIREMENTS
Culture: Fungal culture of 7-10 days old.
Media: Sabouraud agar
Equipments: Sterilized petri dish
Filter paper
Glass rod-u shaped
Slides and coverslips
Lactophenol cotton blue stain
Glass capillary tube
Scalpel
Inoculation needle
Sterile distilled water
Forceps
95% ethanol
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PROCEDURE
Take forceps and with the help of it place a sterilized filter paper in the petri dish. Now keep the sterile unshaped glass rod on the filter paper and put enough distilled water on the filter paper to completely moisten it. Now on the sterile u-shaped glass rod, keep a sterile slide. With the help of scalpel, cut a 5 mm square block of the medium from the plate of Sabourauds agar and transfer this block to the centre of the slide.1 Now in this agar square, inoculate spores or mycelial fragments of the fungus. After inoculation put the sterile cover glass over it. Keep this as a whole in a petri dish and incubate at 37oC for 48 hours. Then after 48 hours, observe if sufficient growth has occurred with hyphae and spores, then proceed for staining. For staining put a drop of lactophenol cotton blue on a clean sterilized microscopic slide. From the slide culture remove the cover glass, and discard the agar block. Now add 95% ethanol to the hyphae on the cover glass. When the alcohol evaporates, place the cover glass on the slide having the lactophenol cotton blue. Now observe it under microscope.
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CONCLUSION
It is one of the rapid method for identification and examination of fungal colonies for its various morphology and characteristics.
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REFERENCES
- Haris JL. Modified method for fungal slide culture. J Clin Microbiol. 1986;24(3):460–1.
