Images of living CHO-K1 cells stably expressing paxillin-EGFP were collected with a CoolSNAP EZ camera with 2 × 2 binning on an Olympus IX71 microscope equipped with a 60×/1.45 NA oil immersion objective. Excitation was from a mercury lamp using a custom EGFP cube from Chroma Technologies (hq480/20×, z488rdc, hq525/50 m). (a) Image corrected for the background intensity of an ROI. (b) Image of the background intensity in a region of the sample with no cells collected with the same parameters as in (a). (c) Image corrected using a normalized background intensity image of (b), followed by subtraction of the average background intensity of an ROI with no cells. The brightness, contrast, and gamma factor are set to the same levels for (a, c), but were adjusted in order to visualize the background intensity in (b). Scale bar is 10 μm.