Posted by
Smith, Mike on
Aug 26, 2005; 3:20pm
URL: http://imagej.273.s1.nabble.com/Non-uniform-illumination-and-noise-reduction-tp3704964p3704966.html
Michael,
I have no way of knowing what your tolerance is for non-uniform illumination, but are your scope and fluorescence system properly aligned? A research grade microscope should have reasonably uniform illumination when properly set up. Apologies if this seems simplistic, but I have found this is often the cause of illumination problems.
Mike Smith
-----Original Message-----
From: ImageJ Interest Group on behalf of Michael Chelen
Sent: Thu 8/25/2005 10:29 AM
To:
[hidden email]
Subject: Non-uniform illumination and noise reduction
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