The Lactophenol Cotton Blue (LPCB) wet mount is most widely used method for the staining and observation of fungi.
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.
- Place a drop of 70% ethanol on a clean microscopic glass slide
- Immerse the specimen in the drop of alcohol
- Add one or at most two drops of the LPCB before the alcohol dries out
- 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
- This preparation is now ready for examination
- Make the initial examination using low power objective. Switch to higher power (40X) objective for more detailed examination of spores and other structures