Background Subtraction

Background subtraction is necessary to accurately quantify the protein material in a spot. It corrects for the intensity level of the scanner bed, for example, and staining variations, etc, across the gel.

In Progenesis SameSpots, the background level for a spot is equal to the lowest intensity value of the image pixels outside the spot's outline. The background level is then subtracted from the intensity value of every pixel inside the spot outline to determine the volume of the protein material in the spot.