Cover Slip Method

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What is Cover Slip Method?

It is a simple, less time-consuming technique that produces high-quality permanent mounts, suitable clinical isolate identification eliminating the potential hazard of infection, student teaching, examination of fungi at different stages of their development without disturbing the arrangement of spores on hyphal structures.

It is advantageous over the slide culture technique that if the first perpetration fails to demonstrate adequate sporulation, there are still three left to be examined t weekly intervals.

The ability of aerial mycelia to adhere to a glass surface has been utilized as a basis of this technique.

Requirements for Coverslip culture Technique

  1. Actinomycete culture
  2. Czapek–Dox agar medium
  3. Sterile petri dish
  4. Inoculating needle
  5. Lactophenol cotton blue

Procedure for Coverslip culture Technique

  1. Pour melted, cooled Czapek –Dox agar medium, 3mm deep in a sterile petri dish.
    Allow the medium to solidify.
  2. Mark four quadrants on the bottom of the plate with a marking pencil.
  3. Transfer a disk of the pure culture of the fungus as spot inoculums in the center of each of four quadrants.
  4. Put a sterile 18mm cover slip cut in to the medium at a 45o angle to te agar surface, passing through each inoculums.
  5. Replace the lid of the petri dish.
  6. Seal the plate with masking tape gently.
  7. Aseptically remove one of the coverslips with sterile forceps.
  8. Remove the masking tape gently.
  9. Reseal the petri dish and reincubate for later examination of the three cover slip cultures every week for four weeks before they are discarded.
  10. Gently heat the cover slip culture in an open flame to fix the culture to the glass.
  11. Put a drop of lactophenol cotton blue on a glass slide.
  12. Gently lower the cover slip culture in to the drop.
  13. Place a second drop of lactophenol cotton blue stain on the cover slip culture.
  14. Cover it with an 18mm square cover slip which makes it a temporary mount for examination.
  15. Put 4-5 drops of histological stain fluid on the mount.
  16. Put 4-5 drops of histological stain fluid on the mount.
  17. Place a second cover slip over the culture which makes it a permanent mount.

References

Further Reading

  1. 10 Instruments Used in Microbiology Laboratory
  2. 8 Qualitative Tests for Protein
  3. Aberration In Lens System
  4. Acid Fast Staining
  5. Algae
  6. Aseptic Transfer Technique
  7. Bacterial Flagella, Fimbriae and Pili
  8. Bacterial Growth and Nutrition
  9. Extremophiles
  10. Fimbriae vs Flagella
  11. Fundamental Microscopy
  12. Growth Curve of Bacteria
  13. MacConkey agar
  14. McFarland Standards
  15. Monochrome Staining
  16. Negative Staining
  17. Ninhydrin Test
  18. Serial Dilution in Microbiology
  19. Spread Plate Technique
  20. Streak Plate Technique
  21. Types of Extremophiles
  22. Xanthoproteic Test