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By Steve Biddle, Sydney Biddle Barrows

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1998). How microtubules get fluorescent speckles. Biophys. J. 75, 2059–2069. , Bokoch, G. , and Waterman-Storer, C. M. (2003). Regulation of leading edge microtubule and actin dynamics downstream of rac1. J. Cell Biol. 161, 845–851. , and Desai, A. (2001). The spindle: A dynamic assembly of microtubules and motors. Nat. Cell Biol. 3, E28–E34. 2. , and Waterman-Storer, C. M. (2004). Fluorescent speckle microscopy of cytoskeletal dynamics in living cells. In “Live Cell Imaging: A Laboratory Manual, D” (L.

And Galjart, N. (2008). Dynamic behavior of GFP-CLIP-170 reveals fast protein turnover on microtubule plus ends. J. Cell Biol. 180, 729–737. , and Duncan, J. (2008). Motion tracking of the outer tips of microtubules. Med. Image Anal. v, 689–702. , Gouveia, S. , Damberger, F. , Bhavesh, N. , van Rijssel, F. , Buey, R. , Winkler, F. , et al. (2009). An EB1-binding motif acts as a microtubule tip localization signal. Cell 138, 366–376. , and Mitchison, T. (1991). Preparation of modified tubulins.

We include a discussion of cell culture and imaging conditions that maintain cell viability. We also provide an extensive discussion of both data collection and analysis that are utilized to estimate the turnover dynamics of MTs. I. Introduction Microtubules (MTs) are major structural components of the cell that are critical for a wide variety of tasks such as motility, cargo transport, spindle assembly, and chromo­ some segregation during mitosis. MTs are composed of a/b-tubulin heterodimers that associate longitudinally into protofilaments, 13 of which associate laterally into the hollow-tube polymer of the MT.

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