Hello ImageJ Listserv,
I am laying out the framework for a colocalization experiment for a researcher here on campus. She has given me a few of her images and I have used them in ImageJ(Fiji) using plug-in Coloc_2. The problem is when I run the images for colocalization... I get an error.
The error states:
Warning! y-intercept high - The absolute y-intercept of the auto threshold regression line is high.
Maybe you should use a ROI, maybe do a background subtraction in both channels.
These results are from a ROI and I tried a background subtraction but it caused other problems.
Before running the test I check the images for Direct information loss, including lossy comprehension and over-exposure. These images passed but I am not sure if they passed the "Incorrect detector offset" test. I was unclear what the histogram log should look like.
I don't know if these are related issues and if they are how I can tell the user to image differently in the Leica software. Thank you for your time.
Best,
Vanessa
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