Some days ago, I built a small program for some of my colleagues to analyse cell images. One minor part of the user interface was the selection of the region of interest. The images are large and need to be analysed in full-size but for the selection, you don't need to see them in full-size. The most direct way to implement this is to use the original image and only show it in a smaller size. With this the original image dimensions are preserved and I can use the original coordinates for the selection of the region. Let me give a small function that does nothing more than showing an image and a rectangular region that can be moved: roiSelector[img_] := DynamicModule[{ pt = Round[ImageDimensions[img]/2], dim = Round[ImageDimensions[img]/6] }, LocatorPane[Dynamic[pt], Show[ HighlightImage[ img, Dynamic@{Rectangle[pt - dim, pt + dim]} ], ImageSize -> 512] ] ] img = Import["http://biology.usf.edu/cmmb/images/cells2.jpg"]; roiSelector[img] ...