6 - STAINING PREPARATION OF FUNGAL SPECIMEN

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9 Terms

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Fungal Stain

Aids in initial diagnosis and identification of fungal infection

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Fungal Stain

Enhance morphological characteristics of fungi under the microscope

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LPCB

most common stain

4
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Potassium Hydroxide (KOH)

Lactophenol Cotton Blue

India Ink

Common Fungal Stains (3)

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India Ink

Stain for C. neoformans

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1, 20% KOH

room temperature, 10-20

denature, keratin, penetrate

KOH Preparation

  1. Prepare two glass slides for skin scraping

  2. Using the first slide, scrape the skin, hair or nails suspected of fungal infection

  3. Collect all the scrapings onto another glass slide

  4. Add _ drop of _ _

  5. Place the coverslip on top of the slide and gently press to get rid of air bubbles

  6. Set aside the specimen at _ _ for _-_ minutes

    • To _ the _ from the tissues and to _ the fungi

  7. Label the slide with type of preparation and initials of the student who performed the procedure

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QUALITATIVE

POSITIVE

NEGATIVE

KOH is a _ identification of fungi. Any presence of any yeast, molds, tubular hyphae, or pseudohyphae is only reported as _. If none, then it is reported as _

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one

Lactophenol Cotton Blue Preparation

  1. Get a colony from fungal culture (see procedure for wet mount preparation)

  2. Add _ drop of LPCB then place a coverslip

  3. Add coverslip on top of the agar block, then gently press it using a rubber stopper to disperse the colony

  4. Label the slide with type of preparation and initials of the student who performed the procedure

  5. Examine all fields under LPO and HPO

  6. Draw and record your observation in the laboratory manual

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urine, serum, CSF

3000, 5

1, 1

50

5-10

India Ink Preparation

  1. Collect _ sample and place it in a sterile urine container

    • India ink can use specimens such as _ and _

  2. Centrifuge the urine sample at _ rpm for _ minutes

  3. Discard the supernatant and save the urine sediments

  4. In a separate test tube, add _ drop of urine sediment and _ drop of india ink

  5. Mix the solution by continuous pipetting or with the use of a vortex

  6. Add _ uL of solution to a clean glass slide

  7. Prepare a smear (follow the blood smear preparation)

  8. Air dry the specimen for _-_ minutes

  9. Label the slide with type of preparation and initials of the student who performed the procedure

  10. Examine all fields under LPO and HPO

  11. Draw and record your observation in the laboratory manual