[Chlamydomonas] Need help to find a working mithramycin staining
protocol for Chlamydomonas
s.kiaulehn from web.de
(by s.kiaulehn from web.de)
Mon Apr 28 10:33:21 EST 2008
Dear Chlamydomonas community,
I am PhD student working on Chlamydomonas reinhardtii and tried to
stain cw15 (mt-) cells with mithramycin, a dye for the nucleus I found
in the Chlamydomonas Source Book. I referred Bridget & Whatley (1975)
to strain the cells by adding 20 µg/ml mithramycin and 5 mM MgCl2
(final concentration). I incubated the cells 20 min at room
temperature and in the absents of light before I watched them through
the microscope (Zeiss Axiophot, 450-490 nm excitation, 510 nm beam
splitter, 520 nm barrier filter).
Unfortunately I was not successful staining the cells. I only could
see strong red auto-fluorescent of chloroplasts but no yellow/green
Is there anybody out there knowing a working mithramycin staining
protocol for Chlamydomonas who is willing inaugurate me into the
secret of staining cells with mithramycin?
Many thanks in advance!
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