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August 2013

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Date:
Thu, 15 Aug 2013 21:31:09 +0800
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Confocal Microscopy List <[log in to unmask]>
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Re: Fluorescent staining of living S. Pombe
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Graham Wright <[log in to unmask]>
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*****
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Hi Laurent,

How about FM4-46 (it will stain the membranes, initially the plasma membrane)? One potential problem though, depending in the rate of endocytosis in your cells, will be that it will be internalised over time if the cells will be in it for a while before imaging. It worked well with Neurospora crassa for us.

Regards
Graham

--
Graham Wright, PhD
Institute of Medical Biology, A*STAR
Singapore

On 15 Aug, 2013, at 20:50, "Rob Palmer" <[log in to unmask]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
> 
> Laurent -
> Calcofluor or other cellulose-binders are typical, caveat being I  
> don't work with yeasts..  A membrane stain called FUN has also been  
> used.  See papers with Ghannoum in the author line.
> Rob
> 
> On Aug 15, 2013, at 5:46 AM, Gelman, Laurent wrote:
> 
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>> 
>> Hello everybody,
>> 
>> 
>> We would need to stain living S. Pombe cells imaged on a  
>> fluorescence microscope to segment thereafter during image analysis  
>> automatically the cells.
>> Staining cell wall and/or membrane would be perfect. Has someone any  
>> specific experience with yeast and/or Pombe?
>> 
>> Note that we need an organic dye and cannot express any recombinant  
>> protein (we are screening a yeast library).
>> 
>> Thanks for your advices!
>> 
>> Best regards,
>> 
>> 
>> Laurent.
>> 
>> 
>> 
>> _____________________________________________
>> Laurent Gelman, PhD
>> Head of Facility for Advanced Imaging and Microscopy
>> (Light Microscopy)
>> 
>> Friedrich Miescher Institut
>> WRO 1066.2.16
>> Maulbeerstrasse 66
>> CH-4058 Basel
>> +41 (0)61 696 43 38
>> +41 (0)79 618 73 69
> 
> Robert J. Palmer Jr., Ph.D.
> Microbial Receptors Section
> Laboratory of Cell and Developmental Biology
> Natl Inst Dental Craniofacial Res - Natl Insts Health
> Bldg 30, Room 207
> 30 Convent Drive
> Bethesda MD 20892
> ph 301-594-0025
> fax 301-402-0396

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