Clearing BG of image with faint florescent signal

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#1

Hi Cell Profiler Community,

I have a cell line that faintly expresses my reporter gene. With a fancy scope I can get the signal pretty clear, but I am collecting images on a GE InCell 2200 and the attached image is what I get after re-scaling the intensity. From this point, I am not sure what I can do to better clean up the image. Before I had more distinction between background and objects so I could get away with some quick and dirty image math. Now I need a bit more finesse.
I can make out the signal so I figure, with the right amount of skill I should be able to clean this up right?

The intensity histogram shows a single hump Gaussian curve so I am not sure how to cleanly discern objects from background. Though since I can visibly see it sort of there has to be a way right?
Example image jpeg snip of Tiff file


Thanks in advance.

Updated so example image would show.


#2

Hi,

I’m not certain which is your signal of interest. My wild guess at the moment would be to try the module “EnhanceOrSuppressFeatures”. If you’re interested in little dots, try option Enhance > Speckles > Size of object-of-interest is about 5 px or less.

If none of the options in EnhanceOrSuppressFeatures would help you, then I suggest to try any other modules that has that kind of “window probe”, where you basically screen the whole image and only enhance the objects that fits into the window-size and ignore the bigger regions, i.e. background.

Maybe that might help. But again, higher quality images should serve you better than these tweaks.

Good luck.