Michael, you might want to try Text2FCS, which converts ASCII data from spreadsheets to the FCS format. The program was created by Joseph Trotter, works nicely and is available at Eric Martz' site of Free Flow Software at http://www.bio.umass.edu/mcbfacs/flowcat.html. - Joerg -----Original Message----- From: Michael Dustin [SMTP:dustin@pathbox.wustl.edu] Sent: Saturday, April 15, 2000 1:23 AM To: Cytometry Mailing List Subject: analysis of overlap/segregation betwee two colors in confocal image data 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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