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Fluorescent method for studying the morphogenesis and viability of dermatophyte cells
Marcia A Lopes1, Olga Fischman, Walderez Gambale
1Instituto de Ciências Biomédicas, Universidade de São Paulo, Brasil.
Mycopathologia
|May 8, 2003
Summary
This study visualizes Microsporum canis development using Fluorescein Diacetate (FDA) and Ethidium Bromide (EB) fluorescence. The method effectively distinguishes viable (green) from non-viable (red) fungal cells during growth stages.
Area of Science:
- Mycology
- Microbial Morphology
- Fluorescence Microscopy
Background:
- Microsporum canis is a common dermatophyte causing human and animal infections.
- Understanding its developmental stages is crucial for effective treatment and control.
- Current methods for assessing fungal viability and morphology can be limited.
Purpose of the Study:
- To investigate the morphogenesis of Microsporum canis using a novel fluorescence-based method.
- To establish a reliable technique for differentiating viable from non-viable fungal cells.
- To detail the distinct growth stages of M. canis in vitro.
Main Methods:
- Culturing M. canis on Sabouraud dextrose agar with cellophane discs for 30 days at 25°C.
- Applying a dual staining technique with Fluorescein Diacetate (FDA) and Ethidium Bromide (EB).
- Examining stained fungal samples under fluorescence microscopy at regular intervals.
Main Results:
- M. canis exhibited five distinct growth stages: tumescence, germination, hyphal development, conidia production, and arthroconidia formation.
- Viable M. canis cells displayed green fluorescence, while non-viable cells showed red coloration.
- Significant morphological changes were observed between day 3 and day 18 of culture.
Conclusions:
- The FDA/EB fluorescence method provides a clear visualization of M. canis developmental stages.
- This technique offers precise differentiation between viable and non-viable fungal cells.
- The study successfully characterized the in vitro morphogenesis of M. canis.