Mitochondrial transport during a growth cone pause. 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 move, one frame for every 8 s is shown. In the movie, the growth cone is toward the right-hand side. The time elapsed is indicated in the top left hand corner. During the first 10 min of the movie, the growth cone advances. It then pauses, but low velocity anterograde transport of the docked mitochondria continues unabated. This movie illustrates low velocity transport continues during prolonged growth cone pauses. 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. Temperature was maintained at 37°C. Movie corresponds to Video 3 in J Cell Biol. 2006 May 8;173(3):373-81.
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