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 *******
