Difficult PI staining of yeast

Miller, Mark A. (mamiller@wista.wistar.upenn.edu)
Tue, 01 Oct 1996 11:21:50 -0400

An associate on mine is having difficulty getting cell cycle profiles of
S. cervisiae. The PI histogram show just one VERY broad peak, and even
that only shows up when I turn the gain and HV on the appropriate FL
detector to their highest settings. It looks a lot like PI exclusion
staining of entirely intact cells.

All of my experience is with mammalian cells. Bruce Milthorpe has
suggested a tertiary ammonium detergent for permeabilization or 0.1 M
HCl for unwinding the histones. Any other suggestions?

Furthermore, has anyone ever seen an article that compares all of the
many ways of getting PI into cells (ethanol, detergents, citrate, etc.?)

Any replies sent to mamiller@wista.wistar.upenn.edu would be truly
appreciated.

Mark Miller

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