Staining of Fungus by using Lactophenol cotton blue

Aim

To stain fungal cells by using Lactophenol cotton blue.

Approach

Fungi are eukaryotic organism and they are classified into two main groups that is yeast and molds.Its cell wall is made up of chitin.Fungal cells have both macroscopic as well as microscopic structure. This microorganism is useful as well as harmful to human being useful because it produces many antibiotics,natural products as well as used in industrial fermentation process.It is harmful in the sense as it causes human diseases,produce toxic substances as well as harm important crops. So it is very important to study fungal species.The branch of science that deals with study of fungal species is called as Mycology.Staining of these fungal cells is a very important step.These are some examples of fungi Rhizopus, Aspergillus, Penicillium, Fusarium, Candida, Mucor etc. Here we are going to stain fungal cells by using Lactophenol cotton blue method this staining technique is also called as mounting of fungus.

Requirement

  1. Young fungal culture
  2. Glass slide
  3. Cover slip
  4. Needle
  5. Lactophenol cotton blue.

Procedure

  1. Take a clean grease free slide.
  2. Add a drop of mounting fluid that is lactophenol cotton blue solution on a slide.
  3. Sterilize the needle and cool it then transfer a mycelial mat on fluid and press it gently so that it easily mix with the stain.
  4. Take a clean cover slip and with the help of a forcep place the cover slip on mycelial mat.
  5. Take a blotting paper and wipe the excess stain .
  6. Observe under low to high power objectives of microscope.

Mechanism

Here in this mounting method we use Lactophenol cotton blue stain where Lactophenol acts as a cleaning agent as phenol is fungicidal in nature so it kills the organism.Lactic acid acts as a preservative and it preserves the structure of fungus.where as cotton blue stains the cytoplasm in light blue colour so here we can observe the fungal structure colourless with light blue background.

Observation

After staining fungus cell we can observe different parts of a cell like conidia, hyphae as well as conidiophore etc.

Result

On basis of observation the type fungal organisms identified.

 

 

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