Fungal staining is a very short and easy method. Here you learn Basic Principle and Procedure of Fungal Staining.
What is fungus?
A fungus is any member of the group of eukaryotic organisms that includes microorganisms such as yeasts and molds, as well as the more familiar mushrooms. Fungi have cells with nuclei. Their cell walls contain chitin, unlike the cell walls of plants, which contain cellulose
Basic Principle of Fungal staining:
Fungal staining is a very short and easy method. There are three processes which we can generally use for fungal staining. These methods are-
- Lactophenol cotton blue (LPCB)
- Periodic acid Schiff (PAS)
- Periodic acid Schiff and Gomari methenamine silver (PAS & GMS) method.
Most widely used method for fungal staining is Lactophenol Cotton Blue staining method. Lactophenol Cotton Blue solution acts as a mounting medium as well as a staining agent by which we can prepare a slide for microscopic examination of fungi. This stain colour the fungal elements as blue in colour.
How you prepare fungal stain in lab?
Lactophenol cotton blue solution is prepared by using phenol crystals (20gm), cotton blue (0.05 gm), lactic acid (20 ml), glycerol (20 ml), and distilled water (20 ml).
What are the functions of Lactic acid, Phenol, and cotton blue?
Lactic acid preserves the fungal structure and also makes the tissue clear.
Phenol acts as a fungicidal or a disinfectant which kills the live organisms and prevents the contamination.
Cotton blue mainly stain the chitin in the fungal cell walls.
Step by step Fungal staining Procedure:
- Take a clean glass slide.
- Add one or two drops of lactophenol cotton blue reagent on a clean and dry slide.
- Transfer a mycelial mat on the fluid through a sterile needle and press it gently so that it easily mixes with the stain.
- Carefully spread the fungal mycelial and make a thin preparation.
- Place a coverslip on the preparation by avoiding any air bubble.
- Blot dries the excess stain.
- Observe under microscope.
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