Re: Live cell imaging prep. for yeast S. pombe
Posted by
Theresa Swayne on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Live-cell-imaging-prep-for-yeast-S-pombe-tp2955849p2957391.html
Graham,
My experience is with cerevisiae, but perhaps it will help. First, we mix agarose with synthetic medium (aka SC). It's less autofluorescent than our yeast extract-based medium (YPD). Second, we use a thinner agarose layer. Our agarose pad is < 100 um thick but the cells are fine for hours.
In our hands, a simple glass-slide mount does not support normal cell physiology for very long (~15 min).
For details, feel free to e-mail me off list, or see
Visualization of mitochondria in budding yeast.
Swayne TC, Gay AC, Pon LA.
Methods Cell Biol. 2007;80:591-626.
PMID: 17445715
Good luck,
Theresa
On May 22, 2009, at 2:01 AM, Graham Wright wrote:
Usually the cells are mounted on an agar plug (agar &
YES media) on a concave slide, covered with a coverslip and sealed.
This works very well for confocal microscopy, but recently upon trying
widefield microscopy we have noticed a significant background
fluorescence, presumably from the agar.
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Theresa C. Swayne, Ph.D.
Manager, Confocal and Specialized Microscopy Shared Resource
Herbert Irving Comprehensive Cancer Center, Columbia University
1130 Saint Nicholas Ave, 222A
New York, NY 10032
212-851-4613