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Tasha,

Don't know about marsupials, but in xenopus when we stain the oocyte  
one cannot see the GV because the DNA is too diffuse so you don't get  
a good signal.  One the chromosomes condense after GVBD they are easy  
to detect.


On Sep 1, 2008, at 10:11 AM, Tasha Czarny wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello
>
> I am trying to stain enzymatically dissociated ovarian follicles to
> determine the oocytes viability using SYBR14 and propidium iodide.  
> I am
> working in a marsupial which has only one layer of tightly bound
> cumuls/granulosa cells and I know from previous experiments that  
> the stain
> can penetrate the egg.
>
> The surrounding cells stain brightly but I cannot find the GV when  
> I assess
> with confocal- either by making a z-stack or manually scrolling.
>
> Does anyone have any hints on how to see the GV, it should be large  
> and
> obvious, but my oocytes appear empty with only staining in the  
> surrounding
> granulosa cells.
>
> Thanks in advance for any help
>
> Tasha Czarny
> PhD student
> Marsupial Research Lab
> The University of Newcastle, Australia

-Khaled

Khaled Machaca, PhD
Associate Professor of Physiology & Biophysics
Associate Dean for Basic Science Research
Weill Cornell Medical College Qatar
Qatar Foundation - Education City
Tel: +974-492-8423   Fax:  +974-492-8422
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