analysis of overlap/segregation betwee two colors in confocal image data

From: Michael Dustin (dustin@pathbox.wustl.edu)
Date: Fri Apr 14 2000 - 18:23:21 EST


We are doing multiple color imaging of molecules involved in T cell
activation.  We need to develop or employ an existing index for the degree
of overlap (or segregation) between two different molecules that are labeled
with different dyes and imaged with a confocal microscope.  One idea would
be to start with a scatter plot (FL1 vs FL2) of the type used for two color
flow
cytometry where individual pixels would be plotted to define regions with
high overlap (on diagonal lower left to upper right) or areas with low
overlap (off diagonal).  We can generate lists of pixel values, but need to
translate the text files to a form readable by flow cytometry software like
Cell-quest to get the plots.  Is there software to go from Excel files to
the FCS format?  Has this problem been dealt with differently by others?
Thanks for any advice.

Michael L. Dustin, Ph.D.
Associate Professor of Pathology
Washington University School of Medicine
660 S. Euclid Ave
Campus Box 8118
St. Louis, MO 63110
Office- (314) 362-9618
Lab- (314) 362-8719
Fax- (314) 362-8888
Jerri Smith, Admin Assist. (314) 362-8740



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