We recently purchased a new fluorescent microscope with a CCD camera.
When capturing images of flourescent beads we have noticed that the
FITC and TRITC signals do not align perfectly when the two images are
merged together. In order to do precise co-localization experiments
we would like to find some software that would take the image files
(usually in 8 or 16 bit TIFF) and be able to have the software
"shift" the red image with respect to the green image. The software
should be able to do this in a precise and exactly repeatable way.
The idea is to emperically determine the "shift" with beads and
then correct this at the software level on our experimental images.
The microscope salesman has some software that will to this but
it is $4000. (ouch!)
I have tried using Photoshop but the only technique I have had success
with involves draging which is neither precise nor repeatable. What I
want is to tell the software to "shift" the red plane 2 pixels up and one
pixel to the left. If anyone knows how to do this in photoshop or other
software that can do this I would be very interested in hearing from you.
chris at biology.wustl.edu