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Module 2 Lab Info

Stack #105900

KOH prep can be used to examine Hair, nails, skin scrapings, fluids, exudates, biopsies
What does the KOH do to the specimen? Dissolves specimen quickly (fungi slowly)
What happens if calcofluor white is added to KOH? Binds to cell wall and fluoresces blue-white under UV light
Historical use of India Ink CSF specimens for C. neoformans
What does India Ink do? Creates black background to visualize capsul
Giemsa, Wright-Giemsa H. capsulatum (intracellular)
Hematoxylin and Eosin (H&E) Pink to pinkish blue
Meyer's Mucicarmine C. neoformans - rose red
Gomori Methenamine Silver (GMS) Fungal elements are black
Papanicolaou stain Pink to blue (cervical, vaginal, some resp samples)
Periodic Acid Schiff (PAS) Red or purple
Time need for yeast to grow 2 to 3 days
Rapid growing mould grows in Less than 5 days
Intermediate mould grows in 6 to 10 days
Slow growing mould takes how long to grow? More than 11 days, sometimes 8 weeks
3 methods used in microscopic examination of moulds Tease/cut preperation, Scotch tape preperation, Slide culture
Tease/cut preperation Organism removed directly from plate, teased apart with teasing needles, can also cut out a piece
Scotch tape preperation Scotch tape pressed onto culture plate, transferred to microscope slide
Slide culture Organism is sub-cultured to small piece of agar, covered with coverslip, grows onto coverslip which is then removed and examined
Best microscopic method for examining moulds is Slide culture
What stain is usually added for microscopic examination? Lactophenol-cotton blue
Hair perforation test Sterile hair floated on sterile water + yeast extract, conidia or hyphae inoculated onto water surface, remove hair shafts and observe weekly for 1 month
Trichophyton hair perforation test results T. rubrum = Neg; T. mentagrophytes = Pos
Urease test Tubes of urease agar are lightly inoculated, incubated 5 days at room temp
Trichophyton urease test results T. mentagrophytes = Pos, T. rubrum = Neg or weak
Trichophyton agars for dermatophyte identification T agar #1 w/o thiamine, T agar #4 w/thiamine; incubate for 10-14 days and observe for growth
T. mentagrophytes growth on Trichophyton agar 4+ on #1 and #4
T. rubrum growth on Trichophyton agar 4+ on #1 and #4
T. tonsurans growth on Trichophyton agar +/- on #1 and 4+ on #4
Rice grain growth Sterile, nonfortified rice grain media; incubate 10 days and observe for growth
Rice grain growth is used to differentiate M. canis (grows) vs. M. audouinii
Created by: blackicecreamcat