KOH and LPCB

• KOH Preparation

  Useful for detecting fungal infection especially dermatophytes which usually resides in skin, hair and nails

  
• KOH Preparation


  1. Add a drop of KOH to a slide

  2. Add the sample

  3. Seal with a cover slip

  4. Gently heat and allow to cool for 15 minutes if the sample contains keratin

  5. KOH breaks down keratin for easier identification of fungi present in the sample
• KOH with Calcofluor white


  Allows better visualization of fungi, the calcofluor white binds with the polysaccharides present in the chitin of the fungus, fungal elements fluoresce apple green depending on the filters used by the microscope

  
• Lactophenol Cotton Blue

  Used to fix tease and tape mounts from cultures, the combination of lactic acid, phenol and blue dye, kills, preserves and stains the organism
• Tease Mount


  Remove mycelium from the media using two needles, place it on the slide with a drop of LPCB stain, add a cover slip and examine microscopically at low and high magnification
• Cellophane tape Mount


  Press cellophane tape onto the surface, place the tape onto the slide with a drop of LPCB stain, examine, advantage is that the conidial arrangement is retained
• Slide Culture


  Useful for demonstrating the natural morphology of fungal structures and encouraging conidiation in some poorly fruiting fungi, can be preserved indefinitely for future comparison
• Standard LPCB mount


  Cultured mold from SDA/PDA are isolated and subjected to teasing and slide microscopy
• Slide Culture


  The agar is directly placed on top of the glass slide thus removing the teasing technique which disturbs the hyphal characteristics, conidial arrangement and fruiting structures
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  • Sporangiospore - asexual spores produced internally within the sporangium (internal spore production)

  
• Allows better visualization of fungi, the calcofluor white binds with the polysaccharides present in the chitin of the fungus, fungal elements fluoresce apple green depending on the filters used by the microscope


  KOH with Calcofluor White
• Used to fix tease and tape mounts from cultures, the combination of lactic acid, phenol and blue dye, kills, preserves and stains the organism

  Lactophenol Cotton Blue