Lactophenol Cotton Blue (LPCB): Principle and Procedure for Fungal Staining

The Lactophenol Cotton Blue (LPCB) wet mount is most widely used method for the staining and observation of fungi.

Principle

LPCB is a stain used for making semi-permanent microscopic preparation of fungi. The LPCB stain has following three components :
Phenol : Kills any live organism.
Lactic acid : Preserves fungal structures.
Cotton blue : Stains the chitin and cellulose of the fungal cell wall intensely blue.

Procedure

  1. Place a drop of 70% ethanol on a clean microscopic glass slide
  2. Immerse the specimen in the drop of alcohol
  3. Add one or at most two drops of the LPCB before the alcohol dries out
  4. Holding the coverslip between the index finger and thumb, touch one edge of the drop of mountant with a coverslip edge and lower gently avoiding air bubbles
  5. This preparation is now ready for examination
  6. Make the initial examination using low power objective. Switch to higher power (40X) objective for more detailed examination of spores and other structures

Result

lpcb-mount

Aspergillus on LPCB Mount

 

About Dhurba Giri
Dhurba Giri is the author at LaboratoryInfo.com, a scientific blog dedicated for Medical Laboratory Professionals. He's a fresh graduate of Bachelor of Science in Medical Laboratory Technology (B.Sc.MLT) from Pokhara University, Nepal. Connect with him on Facebook !

2 Comments on Lactophenol Cotton Blue (LPCB): Principle and Procedure for Fungal Staining

  1. The above genus, I believe, is Penicillium and not Aspergillus. Please check.

  2. it is nice to read. I can remember easily.
    thank you.

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