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Image analysis problem - (Oct/16/2007 )


Hope somebody out there can help me with this one. I’m supposed to compare fluorescence intensity (PI incorporation) between different images... problem is I do not have a camera incorporated with the microscope, but I take pictures from the oculars. So it is impossible for me to set exposure on the control condition and I end up with different background intensity anyway between images, which alters final analysis. So here’s the question: is there a way to somehow normalize pictures with an image analysis software after they’re taken? What would be the procedure? I’m now using Image J software.

Thanks to anyone who can help!


anything you do after the fact is manipulation of the image - your set up means you can judge for yourself a little but publishing would be unlikely. (if you alter the background the intensity of positive labelling will change thus affecting the very fluorescence intensity you are trying to measure)
bite the bullet and get a proper camera system or you are just wasting your time