Introduction of Smart Background Correction in CoLocalizer Pro 5.2 gave us the reason to recall the changes the Background Correction panel underwent during development. These changes can be summarized as “from raw to complex to simple”.
It started with a simple standard panel containing channel selector as well as Auto and Manual modes for correction. Auto mode had 3 Image Pattern presets depending on the appearance of images. Manual mode offered correction using either Background ROI or Threshold Value. Background ROI was simple yet very efficient: users were selecting small areas of actual background on the images and then subtracted these selected values of pixel intensities from them. The Threshold Value was a slider capable of removing all pixels below the desired value. This approached worked too, but wasn’t taking into consideration the properties of images.
Version 3 modified channel selector to make it a channel pair selector and added histogram for visual feedback. It still had two tabs for selection, which some users found a bit confusing.
Finally, version 5.2 introduced Smart Background Correction. This feature dramatically improved the way the background is handled. With this approach, background correction and ROI selection are tied into a single step as the background is corrected specifically for the selected ROI considering its unique properties. This represents a major step in improving the reliability of colocalization quantification because it allows comparison of the results from different experiments.
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