I have a colleague in my department that has seen some of the great images
that have been taken in Tetrahymena with GFP tagged strains and had the
following question about his problem with autofluorescence with his FITC
conjugated antibody images.  If anyone has any suggestions feel free to
email him directly.


Josh Smith

Josh Smith, Ph.D.
Assistant Professor
Missouri State University
Department of Biomedical Sciences
Professional Building Rm. 400
901 S. National Ave.
Springfield, MO  65897
Phone: (417) 836-5321
Fax: (417) 836-5588
Email:  [log in to unmask]

Mike Craig wrote:
³Microscopists, help! A particular cell line with which I am working is
demonstrating autofluorescence in the green range. I am using FITC as the
fluorochrome and an Olympus confocal microscope for imaging. The
autofluorescence is detectable in the cytoplasm (both diffusely in some
cells and at vesicular foci in others) at both 10ms and 20ms exposure times,
and the raw images are stored in TIF format. My question is this:  Can
anyone suggest an image analysis protocol, applied to the TIF images,
whereby the signal due to autofluorescence can be subtracted from, or
discriminated from, the signal due to the FITC labeled entity?²

If you have any suggestions please email him at
[log in to unmask]