DAPI staining

[SLForsburg]

Hoffmacs at HERMES.BC.EDU ("Hoffmacs") wrote:
>Hi fellow netters,
>I want to photograph some S. pombe cells, both phase and DAPI-stained.  Is
>there a fast and simple method for fixing and permeablizing cells for
>DAPI-staining relative to the procedures used for immunofluorescense?  Thanks
>in advance.
>Charlie Hoffman

Hi Charlie.  You can live stain pombe with DAPI if you are in a REAL hurry (Mix cells with DAPI, mount).  Or, you can heat fix them (put 10ul culture on a slide, put it on a hotplate until the liquid just evaporates, add DAPI and mounting medium and coverslip).  I like to ethanol fix them so I can keep them for a while (harvest cells, resuspend in cold, 70% ethanol and chill.  Before use, wash once in PBS, then either stain before mounting or place onto DAPI in the mounting media;  use poly L lysine coated cover slips so they dont float away).

Give me a call or an email if you need more details.

cheers
susan

susan_forsburg at qm.salk.edu