Staining Procedure for Rapid Identification of Fungi

Scrapings from involved ocular site complemented with the appropriate staining method can offer the ophthalmologist circumstantial as well as definitive information concerning the identity of the invading organism

Fig. 1: KOH pellets and preparation of 10% KOH

Fig. 4.2: Hyaline, septate fungal filaments in 10% KOH wet mount of corneal scraping specimen (400 X magnification)
Fig. 2: Hyaline, septate fungal filaments in 10% KOH wet mount of corneal
scraping specimen (400 X magnification)

Fig. 3: Hyaline, septate, large, branching fungal filaments of corneal scraping in 10% KOH wet mount (400 X magnification)
Fig. 3: Hyaline, septate, large, branching fungal filaments of corneal
scraping in 10% KOH wet mount (400 X magnification)

Fig. 4: Hyaline fungal filaments in KOH wet mount (400 X magnification)
Fig. 4: Hyaline fungal filaments in KOH wet mount (400 X magnification)

Fig. 4.5: Hyaline, septate fungal filament in KOH wet mount of aqueous fluid (400 X magnification)
Fig. 4.5: Hyaline, septate fungal filament in KOH wet mount of aqueous fluid (400 X magnification)

Fig. 4.6: Pigmented, septate fungal filaments in KOH wet mount of corneal scrapings.  Culture grew Curvularia (400 X magnification)
Fig. 6: Pigmented, septate fungal filaments in KOH wet mount of corneal scrapings.
Culture grew Curvularia (400 X magnification)

Fig. 7: A KOH mount showing hyaline septate fungal filaments and thin Nocardia filaments (isolated from a patient with corneal ulcer caused by mixed infections involving fungi and Nocardia)—(400 X magnification)
Fig. 7: A KOH mount showing hyaline septate fungal filaments and thin Nocardia filaments (isolated from a patient with corneal ulcer caused by mixed infections involving fungi and Nocardia)—(400 X magnification)

Fig. 8: Thin broken fungal filaments from a corneal ulcer under antifungal treatment (400 X magnification)
Fig. 8: Thin broken fungal filaments from a corneal ulcer under antifungal treatment (400 X magnification)

Oval shaped yeast cells in KOH wet mount (400 X magnification)
Fig. 9: Oval shaped yeast cells in KOH wet mount (400 X magnification)

Fig. 10: Gram’s stain kit
Fig. 10: Gram’s stain kit

Fig. 4.11: Gram’s stain showing branching fungal filaments from corneal scraping (1000 X magnification)
Fig.11: Gram’s stain showing branching fungal filaments from corneal scraping (1000 X magnification)

Fig. 12: Large fungal filaments stained with crystal violet of Gram staining (1000 X magnification)
Fig. 12: Large fungal filaments stained with crystal violet of Gram staining (1000 X magnification)

Fig. 4.13: Long fungal filaments with sickle shaped spores  (morphologically resembles Fusarium spp) from corneal ulcer cases (1000 X magnification)
Fig. 13: Long fungal filaments with sickle shaped spores
(morphologically resembles Fusarium spp) from corneal ulcer cases (1000 X magnification)

Fig. 14: A Gram’s stain slide picture showing fungal filaments (½) and gram-positive bacilli (isolated from a patient with corneal ulcer caused by mixed infections involving fungi and bacteria) (1000 X magnification)
Fig. 14: A Gram’s stain slide picture showing fungal filaments (½) and gram-positive bacilli (isolated from a patient with corneal ulcer caused by mixed infections involving fungi and bacteria) (1000 X magnification)

Fig. 15: A Gram stain slide picture showing fungal filaments and gram-negative bacilli (¾) ( isolated from a patient with corneal ulcer caused by a mixed infections involving fungi and bacteria) (1000 X magnification)

Fig. 16: A Gram stain slide picture showing fungal filaments and gram-positive beaded filaments of Nocardia ( isolated from a patient with corneal ulcer caused by mixed infections involving fungi and Nocardia) (1000 X magnification)

Fig. 17: A Gram stain slide picture from corneal scrapping showing gram-positive  budding yeast cells of Candida spp (1000 X magnification)
Fig. 17: A Gram stain slide picture from corneal scrapping showing gram-positive
budding yeast cells of Candida spp (1000 X magnification)


Fig. 18: A Gram stain slide picture from corneal scrapping showing clumps  of gram-positive yeast cells (100 X magnification)
Fig. 18: A Gram stain slide picture from corneal scrapping showing clumps
of gram-positive yeast cells (100 X magnification)

Figs 19A and B: Fungal elements stain silver to black in Gomori  methenamine silver (GMS) stain – Tissue biopsy (400 X magnification)
Figs 19A and B: Fungal elements stain silver to black in Gomori  methenamine silver (GMS) stain – Tissue biopsy (400 X magnification)
Figs 19A and B: Fungal elements stain silver to black in Gomori
methenamine silver (GMS) stain – Tissue biopsy (400 X magnification)





Figs 20A and B: Blue white fluorescence of fungal hyphae of
corneal scraping in KOH Calcofluor staining (A – 200 X magnification, B – 400 X magnification)
Figs 53A and B: Scedosporium prolificans - Basally inflated annllides (→)
Figs 53A and B: Scedosporium prolificans - Basally inflated annllides (→)


This is only a part of the book : Aravind's Atlas of Fungal Corneal Ulcers 1st Edition of authors: Lalitha Prajna MD, Vijayakumar MSc, N Venkatesh Prajna DNB and M Srinivasan MS. If you want to view the full content of the book and support author. Please buy it here: https://goo.gl/kz9puC

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Staining Procedure for Rapid Identification of Fungi
Scrapings from involved ocular site complemented with the appropriate staining method can offer the ophthalmologist circumstantial as well as definitive information concerning the identity of the invading organism
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