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Laboratory Methods

	Antifungal Susceptibility Testing

	Culture Techniques and Media

	Microscopy Techniques and Stains

	Specimen Collection and Processing

School of Molecular & Biomedical Science
THE UNIVERSITY OF ADELAIDE
AUSTRALIA 5005

Contact:
Dr David Ellis

Email

Telephone:
+61 8 8161 6459
Facsimile:
+61 8 8161 7589

Potassium Hydroxide - Dimethyl Sulfoxide (KOH-DMSO) Preparation.

For the direct microscopic examination of skin scrapings, hairs and nails for fungal elements.

Method:

Using a safety pipette, add DMSO to the distilled water and then dissolve the KOH in solution.

Dimethyl sulfoxide (DMSO) 40 ml

Distilled Water 60 ml

Potassium hydroxide 10 gm

Making mounts for microscopy:

1. Using an inoculation needle remove a small portion of the specimen and mount in a drop of KOH-DMSO on a clean microscope slide.

2. Cover with a coverslip, squash the preparation with the butt of the inoculation needle and then blot off the excess fluid. Do not heat the preparation.

3. The mount must be examined within 20 minutes. Using low light, examine microscopically for the presence of unstained refractile fungal elements.

Comments: DMSO gives more rapid maceration and clearing than potassium hydroxide alone, however preparations do not keep.