Dual-channel spinning-disk confocal time-lapse movie showing kinetochores (Alexa488 labeled CENP-A antibody) in metaphase (red) relative to poleward microtubule flux. Fluorescent speckle microscopy (FSM) of microtubules and kinetochore imaging during metaphase was performed as described in Maddox et al. (2002, 2003), and with a 100X/1.4 NA Plan Apochromat objective (Nikon) with 2X2 binning in the cooled CCD camera (model ER; Hamamatsu Corporation). MetaMorph® software (Universal Imaging Corp.) was used to control shutters, wavelength selection, image acquisition, and storage. Images were acquired at 5-s intervals. Sequential images of different fluorophores were acquired at 3-15s intervals, depending on the experiment. Paired images from different color channels were within 0.5 s of each other. Movie is video 2 from J Cell Biol 2003 162:377-382.
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