Re: Live cell imaging prep. for yeast S. pombe

Posted by Jörg Bormann on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Live-cell-imaging-prep-for-yeast-S-pombe-tp2955849p2956014.html

Hello,
for live-cell imaging of (filamentous) fungi we use agarose instead of agar. This diminishes autofluorescence to some extent. Polylysine didn´t work that well with filam. fungi.
Best wishes - Joerg

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Jörg Bormann
Westfälische Wilhelms-Universität Münster
Institut für Botanik
Schlossgarten 3
48149 Münster
Phone: [phone]

2009/5/22 Graham Wright <[hidden email]>

Hello,

We are doing some live cell imaging of Schizosaccharomyces pombe
(fission yeast). 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. Simply mounting the cells in
liquid media on a normal glass slide significantly removes the
background problem, but creates another - the cells do not adhere to
the glass and so make long term timelapse imaging very difficult.

So, can anyone suggest ways of encouraging the cells to adhere that
work well for yeast, particularly S. pombe? Alternatively any
suggestions of agar, or other gelling agents that do not fluoresce
would be appreciated.

Thanks in advance for your help,
Graham

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