Microincubator

After transfection, the dishes containing the attached cells were placed on a Harvard microincubator located on top of the stage of a fully motorized Nikon TE-2000 E inverted microscope equipped for differential interference contrast, epifluorescence, and TIRFM. For conventional time-lapse fluorescence microscopy, cells were imaged using a 60X or a 100X oil-immersion objective, an ORCA II ER camera (Hamamatsu) and MetaMorph software (Molecular Devices) [32 ].