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Calcofluor staining of bud scars

Desrochers Normand desrocn at ERE.UMontreal.CA
Tue Aug 16 11:36:08 EST 1994


	I'm trying to visualize bud scars on S.cerevisiae. The 
technique I'm using starts with an o/n culture. 1ml of cells are 
harvested, resuspended in calcofluor (1mg/ml) and incubated at room 
temperature for 5 minutes. Finally, the cells are washed 3 times with 
water and analized on a fluorescent microscope (with the appropriate 
filter).

	The results are appreciable and I can see some bud scars, but I 
wonder whether someone know a better way to carry out the staining. Is 
there's a way to increase the contrast between the scars and the cell 
membrane? Is it better to start with an old culture (to see more scars on 
the cell) or with a log phase culture? Is the calcofluor concentration is 
appropriate? Etc.

	Any suggestion will be greatly appreciated!

      ********************* DESROCHERS, Normand **********************
      **************** desrocn at alize.ere.umontreal.ca ****************
      ************** dep. Microbiologie et Immunologie ***************
      ******* Universite de Montreal (514) 343-6111 poste:1432 *******



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