The program started band fluorescence quantification by analyzing a single line of pixels running from the upper to the lower bound and did so sequentially for each line of pixels starting from left to right covering the width of the lane. For each line of intensity data, the MatLab function "msbackadj" was used to flatten the uneven background created by the lighting source. This created a flat background level at zero fluorescence intensity from which the band peaks could be detected along the length of the defined line.