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Judy,

"Colocalization Analysis"  can mean everything and nothing.... You  
should ask the investigator to explain to you what his biological  
question is, preferably in plain English.... this will determine  
which method is most appropriate. Does (s)he want to know the  
proportion of cells that have all three proteins? the proportion of  
cells with protein A, that also have protein B and C?  Does he want  
to know in any given cell how many foci are present containing all  
three proteins? or maybe what fraction of foci of Protein A are  
colocalized (within what distance?) with foci of B and C?  The  
percentage of the cell's area/volume containing all three proteins  
(above what threshold?), and so on...

Without a specific question you can't provide a specific answer...

--
Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA 98109-1024


http://www.fhcrc.org/science

On Aug 27, 2008, at 6:58 AM, Judy Trogadis wrote:

> Hi, Michael,
>
> You're right, 2 runs would do it - but one of the users of our  
> imaging facility has triple labelled preps ready to analyze. He is  
> looking for the presence of 3 proteins in a cell but I am not sure  
> about their proximity to each other. Visual observation at high  
> magnification could give a clue. I think the user wants some  
> numerical value for a grant.
>
> Thanks.
> Judy