I am processing some of my images. Normally I start aplying some gaussian filtering, then I subtract the spatial variations of the background intensities using rolling ball function and afterwards I subtract some background signal by thresholding it.
In some of my images that have very low signal, I have observed, that if before all of that, I enhance contrast “auto” and apply LUT, I got much better imaging results (the signal that comes from my cells is better distinguised from the background noise) than when I do not do it. But I cannot understand why. Does anyone have any idea? Is it correct to apply auto contrast enhancement and apply LUT at the beginning of the processing?