Re: background removal in fluorescence

Posted by Martin Wessendorf on Sep 20, 2005; 7:11pm
URL: http://imagej.273.s1.nabble.com/background-removal-in-fluorescence-tp3704817p3704821.html

Dear ImageJ-ers--

I realize that I didn't articulate the problem particularly well.  --We
want to make some montages of images.  Our illumination is not perfectly
even and so parts of the field are illuminated more brightly than
others.  Thus when we try to make the montage, there are abrupt
inconsistencies in the lighting across it.

I had thought that the following *should* work:

1) Take an image of a uniform fluorescent field (image A)

2) Take an image of the specimen (image B)

3) Divide B by A; multiply by the mean intensity of A.

(--I think that this is what Wayne's plugin does.)

However, when we use that method, the inconsistencies of the
illumination are greatly accentuated rather than being reduced.  My
guess is that this is due to non-linearity of response in our CCD camera.

I'd be delighted if anyone has a work-around for this problem!

Martin
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                E-mail: [hidden email]