I need to study the cell morphology of a fungal hyphae, specifically cell wall and cell membrane. Which technique is better - confocal microscopy or Total internal reflection fluorescence microscopy (TIRFM)? And which dye is best suited - Calcofluor white or Trypan blue, as both have been considered equally desirable for the purpose.
Also, can anybody who has had some experience in fluorescence microscopy, provide a tested and reliable protocol for sample preparation and imaging?
Thanks a lot!
You should try the use of propidium iodide for cell wall imaging, works fine with CLSM and also with fluorescence microscopy in plants.
Regards
I agree with Aarón but be careful with propidium iodide because in my experience, it only stains the damaged cells because it is highly membrane impermeant. Syto9® is a reliable dye for any cell as it is highly membrane permeant.
Regards
We have a paper describing what you are asking for. Look for Mora-Montes et al. 2011 infect. Immun;79(5):1961-70. PI won't work well in Aspergillus intact cells.
Thank you Mr. Barraza, Mr. Long and Mr. Mora for you kind and prompt suggestions. Thanks again.
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