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Dear Dorte,
TP3 will sometimes bleach quickly unless you use at least 4 µM and low  
excitation power.  Yo-Pro-1 seems more robust at 4 µM.  The Lo-Pro-1  
(red) seems fairly robust, but tends to show up weakly in the green  
channel.

Regards,
Glen
On Jul 29, 2005, at 3:41 AM, DORO (Dorthe Rønn Olesen) wrote:

> Search the CONFOCAL archive at  
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello all.
>
> I have a quick question for those of you with experience in doing  
> confocal microscopy of fixed cells cleared with BABB.
>
> I have done whole mount immunocytochemistry of large aggregates of  
> mouse ES cells (embryoid bodies) with good results after clearing, but  
> I find that a nuclear counterstain with DAPI or Hoechst bleaches  
> extremely fast after clearing with BABB. I work on a Zeiss LSM 510  
> Meta using a 405 laser for excitation of DAPI and Hoechst.
>
> So my question is if other people has had similar problems when using  
> these dyes and, if that is the case, if anyone has any experience with  
> other nuclear counterstains that are not sensitive to clearing,  
> perhaps with TO-PRO-3 or PI?
>
> Alternatively does anyone know of other clearing agents that are  
> compatible with DAPI or Hoechst counterstains?
>
> Thank you
>
> Dorthe
>
> ____________________________________
>
> Dorte Rønn Olesen, DVM
> PhD-student
>  Developmental Biology
>
>  Hagedorn Research Institute
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>
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
(206) 616-4156
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