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Dear Jennifer,

I don't know any reference but I would do the following quick (and dirty 
depending on the applications) approach.

1. threshold the fluoresce and plot it over time or z. You can use on 
imagej image->stack->plot z axis profile
2. with that you should get a exponential like decay curve that you can 
  use as a correction factor for each slice
3. The resulting intensities would be
  In=I+I(1-Cn)
where I is corrected intensity for the frame n, I is the real picture 
and C the correction factor for the specific frame n.

Please note that you will have worth statistics with higher n since 
there is less SNR.

Regards,
NM

Jennifer Waters wrote:
> Search the CONFOCAL archive at 
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>  
> Good morning fellow microscopists.  If you haven't heard, Kevin Hodgson 
> is apparently out of the office.  :)
>  
> Does anyone know a good reference for how to measure and correct for 
> photobleaching when quantitating fluorescence over time or in a 3D stack?
>  
> Thanks, Jennifer
> 
> -- 
> Jennifer Waters, Ph.D.
> Director, Nikon Imaging Center at Harvard Medical School

-- 
Nuno Moreno
Cell Imaging Unit
Instituto Gulbenkian de Ciência
http://uic.igc.gulbekian.pt
http://www.igc.gulbekian.pt
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