The program would then define the physical dimensions of band in the Y direction by defining the fullwidth-half-max of the peak. The area under this defined portion of the curve was calculated using the trapezoidal approximation using the MatLab function "trapz". This process was repeated for the next line of pixels until the entire width of the user defined lane was analyzed. The X dimensions of the band were determined by setting a lower threshold limit for peaks to be defined by using a "Min Peak Prominence" of 1500 for DOX. The area under the curve defined by each line of pixels was added together to produce a total intensity for the fluorescence band.