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Ultrastructure of Trichophyton mentagrophytes stained with neutral red
1Department of Dermatology, Keio University School of Medicine, Tokyo, Japan.
Abstract:
The ultrastructure of Trichophyton mentagrophytes cells stained with neutral red was investigated using electron microscopy and X-ray microanalysis. Fixatives containing molybdenum and chromium were used to prevent the outflow of neutral red. Electron-dense particles composed of metals and dye were observed exclusively in the vacuoles, which were increased in number and size, but not near the cell wall. Results indicate that neutral red passes directly through the fungal cell wall and is incorporated into the vacuoles.
Insights
Neutral red dye enters Trichophyton mentagrophytes cells directly through the cell wall. The dye accumulates within the fungal vacuoles, increasing their size and number.
Area of Science:
- Mycology
- Cell Biology
- Biophysics
Background:
- Trichophyton mentagrophytes is a common fungal pathogen.
- Understanding fungal cell permeability is crucial for developing antifungal treatments.
- Neutral red is a vital stain used to visualize cellular structures.
Purpose of the Study:
- To investigate the cellular uptake mechanism of neutral red in Trichophyton mentagrophytes.
- To determine the specific cellular compartments where neutral red accumulates.
- To elucidate the interaction of neutral red with fungal cell structures.
Main Methods:
- Transmission electron microscopy (TEM) was employed to examine cellular ultrastructure.
- X-ray microanalysis was utilized to identify the elemental composition of stained particles.
- Specialized fixatives containing molybdenum and chromium were used to preserve neutral red within cells.
Main Results:
- Electron-dense particles, identified as metal-dye complexes, were exclusively found within vacuoles.
- Vacuoles within the fungal cells were observed to increase in both number and size.
- Neutral red accumulation was not detected near the fungal cell wall.
Conclusions:
- Neutral red permeates the fungal cell wall of Trichophyton mentagrophytes directly.
- The dye is sequestered within the vacuoles, leading to their enlargement.
- This uptake pathway suggests vacuoles play a key role in neutral red accumulation in this fungus.