Description

Chicken DRG neurons were grown on coverslips coated with laminin/poly-L-ornithine for 2 d and stained with 0.1 µM MitoTracker red CMXRos. Images were acquired every 2 s. To reduce the size of the video, one frame for every 10 s is shown. In the video, both DIC and fluorescent images are shown. The growth cone is toward the right-hand side. The time elapsed is indicated in the top left hand corner. During the process of growth cone advance, docked mitochondria moved at a low velocity in the anterograde direction in a coherent manner. A kymograph of the movie is shown at the bottom of the screen. Time is on the y axis and distance is on the x axis. The moving bar on the right hand side of the kymograph marks the time position. This video illustrates how kymographs correspond to time-lapse movies of mitochondrial transport, Bar, 10 µm. Images were acquired on a confocal system (FV300 FluoView; Olympus) on an IX70 microsocpe using a 60×, 1.4 NA, PlanApo objective. Samples were illuminated with an argon/krypton Omnichrome 488/568/647 line laser set to standby mode, with the laser intensity set at 6%, the confocal aperture set at 5, and pixel dwell times of either 100 or 200 microsec; the light was further attenuated with neutral density filters (either 25 or 50%). Cells were observed in an enclosed flow chamber maintained at 37°C. This video corresponds to Figure 1 and video 1 from J Cell Biol. 2006 May 8;173(3):373-81.