Posted by
Joel Sheffield on
Aug 25, 2005; 4:36pm
URL: http://imagej.273.s1.nabble.com/Non-uniform-illumination-and-noise-reduction-tp3704964p3704972.html
Can you get an image of the illuminating field without the sample?
If so, you could subtract, or divide (depending on what works for
you)this image from each of the frames, and then adjust b/c.
Alternatively, you might be able to create a pseudo background by
taking one of your original images and carrying out a blur operation
to reduce the effect of the sample, but retain the broad intensity
distribution.
Joel
> Hello,
> I'm trying to analyze some calcium dye fluorescence images. The
> illuminating field appears to be non-uniform, and I'm looking for a
> good way to calculate and remove the background illumination. So far,
> I've tried both the built-in "Subtract Background" command (Process ->
> Subtract Background) and the plugin "Background Correction"
> (
http://rsb.info.nih.gov/ij/plugins/background.html). These both to
> seem to work reasonably well, but the second does not work for a
> multiframe image. I was wondering if anyone had any experience with
> these two tools, and if they could provide any insight into how these
> tools work, and if there are others I should be considering.
>
> Because the image set is a 500 frame t-series, it seems like the best
> algorithm would be one that uses the data from every frame to
> calculate the best possible approximation of the illuminating field.
> Additionally, such a utility could identify likely points of noise
> because the noise will change much more rapidly (from frame to frame)
> than the actual signal. Is there anything like this for ImageJ that
> anyone can recommend? I would really appreciate any suggestions or
> insight.
>
> Thanks,
> Michael Chelen
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
[hidden email]
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs