Protease charge changing assay software analysis.

This script will take an image where the wells are marked with white lines and will quantify the fluorescence pixel intensity of the signal using an automated integration methodology. There are a few steps on this script. The first step is to load the images in tif file and the script will convert them automatically into png file. Next, it will compute an image gradient in both x and y direction and merge them into a single image to be able to detect any borders that are present in the image. After, the script will recognize the y pixel where the wells begin (where the gel start) and the pixels of x where every well starts and ends. After finding the wells, the script will take the average 20 pixels of the center of every well and average them. After averaging them, we will plot a density histogram with the average 20 pixel intensities for all pixels in the y axis. Next, a signal integration using the Simpson rule will be perform by also taking into account how deep into the gel every specific probe runs. Finally, the integration results will be plot into a histogram where values are indicated and original gel image will be returned with a rectangle around the areas where the signal was integrated for every well.