Staining fungal nucleoli

[Austen]

Hi!

I am attempting simple 'ole light microscopy, looking for nucleoli in my 
fungi, but so far haven't met with the greatest success.

I have been using smears of fungal protoplasts that I fix in either 
methanol or Carnoy's fluid (1 acetic acid:3 ethanol), and then stain 
with 0.2% methylene blue mixed with saturated sodium borate.  
Occasionally I get a few nuclei that stain up with nucleoli (or what I 
presume are nucleoli), but its pretty inconsistent, and only a very few 
cells stain properly.

Does anyone have any experience with this sort of work (especially with 
fungi), or any alternative methods (I don't want to go into antibody 
work or anything like that), as this isn't really my area.

E-mail to me.

Cheers,


Austen   <-:)