Human retinal pigmented epithelial (RPE) cells labeled for clathrin-coated pits (green), focal adhesions (red) and nuclei (blue). RPE cells stabily expressing 'clathrin light chain a' tagged with EGFP (EGFP-LCa) were plated directly on glass coverslides. After 72 hrs, cells were fixed for 2 min in a 1:1 mixture of 2% paraformaldehyde/0.5% Triton X-100, followed by a 30 min fixation in 4% paraformaldehyde. Cells were then stained with mouse monoclonal antibodies to paxillin, detected with goat anti-mouse Alexa 568. Nuclei were stained with DAPI. Image was collected on an Olympus IX71 inverted microscope coupled to a Hamamatsu ORCA-ER digital camera using a 60x 1.42 NA Plan Apo N objective (Olympus). The exposure times were 200, 500 and 70 msec for the red, green and blue channel, respectively. A xenon-arc lamp (Lambda LS stand-alone illuminator; Sutter Instrument) was used with adequat filters (Chroma). Bar: 5µm