I'm interested in quantitation of protein expression in
tissue slices & cell cultures using immunofluorescent probes. The
protein in question is a mitichondrial membrane protein.
My method consists of a fluorescent microscope + CCD and image
processing & analysis software (IPPlus for MS-Windows ver. 1.1)
I'm having trouble segmenting (separating from the background)
the mitochondria - a high error margin due to artifacts, clustered
cells etc. There is a resolution problem, obviously, and one of the
things I'm looking for is a way to overcome that via filtering, etc.
If anyone out there is familiar with this type of work- namely
quantitative immunofluorescent microscopy, and can recommend techniques
or publications in which these techniques appear, her/his input
would be most welcome.
PLEASE DO NOT:
1) Tell me about the NIH Image (seems like an automatic response here
for people with image-analysis problems). Not suitable for my platform,
and I'm satisfied with what I've got, softwarewise and hardwarewise.
A good *procedure* is what I need.
2) Tell me about Methods in Cell Biology Vols. 29&30. I've read them.
Would like info besides.
Thanks in advance,
PS Anyone familiar with Media Cyberntics ImagePro Plus, and would like to
discuss general aspects of that, I'll be More than happy.