DAPI staining

[Michael Lichten]

>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
>"I'd rather be fission."
>P.S.  When trashing the "Hoffman and Winston" protocol for plasmid rescue
>from yeast, please refer to it as the "Smash and Grab" technique.  Thank you.


DAPI Staining of yeast nuclei

1.      Pellet cells, resuspend in 50 ul water.  Add 1 ml ETOH.
Alternatively, fix cells in 50% ETOH ahead of time and chill at -20 C.

2.      Add DAPI to 0.1-0.2 ug/ml.  Let sit 5 min.

3.      Spin out, rinse 2x with water.  Resuspend in a small volume
(usually 20-50 ul).

Is that fast enough?



Michael Lichten
lichten at helix.nih.gov