Slide culture technique of fungi


 
As the structural components of molds are very delicate, they can be distorted very easily. So, the slide culture technique is used to minimize the chances of distortion. Preferably, deep concave slide deposited with suitable acidic media is used. The media is covered by coverslip and incubated in a moist chamber. In this way, we can carry out direct microscopic identification of the culture. Molds are identified by spore shape and type, type of mycelium, and type of sporangia.

Materials

  • Fungal culture (7-10 days old)
  • Sabouraud agar
  • Bunsen burner
  • Water bath
  • Concave glass slide
  • Coverslips
  • Pasteur pipette
  • Petri plates
  • Inoculation loop/needle
  • U shaped bent glass rod 
  • Filter paper
  • Petroleum jelly

Procedure 

  1. Melt the sabouraud agar by keeping it in boiling water, then maintain the temperature at 45°C by keeping it in a water bath.
  2. Place a piece of round cut filter paper in the Petri plate
  3. Place sterile bent glass rod in the Petri plate.
  4. Sterilize the glass slide and coverslip by dipping in the ethyl alcohol and then burning the alcohol over the flame.
  5. Place the glass slide over the bent glass rod in the Petri plate with a concave surface upwards.
  6. Cover three sides of concavity with the petroleum jelly by the toothpick and left open the 4th side for air passage.
  7. With the Pasteur pipette place one to two drops of Sabouraud agar in the concavity of the slide.
  8. Place a coverslip over the slide and allow it to solidify.
  9. When the agar is fully solidified, slide the coverslip to one side with the forceps.
  10. Inoculate the agar with the inoculation loop or needle from the stock culture.
  11. Gently push the coverslip to its original position.
  12. Moisten the filter paper by with sterile water.
  13. Place the slide over u shaped bent glass rod and incubate for 5-7 days at 25°C.

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