By Lynne Cassimeris, Phong Tran
This volume contains chapters through specialists world wide on many elements of microtubule imaging in dwelling and glued cells; assays to check microtubule functionality in a wide range of version organisms and cultured cells; excessive solution methods to check of the cytoskeleton. The authors percentage their years of expertise, outlining power pitfalls and important components to contemplate in experimental layout, experimental implementation and information interpretation.Iincludes chapters via specialists world wide on many features of microtubule imaging in dwelling and glued cells; assays to check microtubule functionality in a wide range of version organisms and cultured cells; excessive answer ways to review of the cytoskeleton. The authors percentage their years of expertise, outlining strength pitfalls and demanding components to think about in experimental layout, experimental implementation and information interpretation.
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Extra info for Microtubules: in vivo
Typically adds several hundred milliseconds of sample exposure before and after image acquisition, which results in significantly faster photobleaching especially at low exposure times. In addition, it is important to use optimized emission filter sets. For example, for single-channel EGFP imaging, we use a long-pass dichroic mirror and emission filter (Semrock) with edge wavelengths of $500 nm. This allows maximum detection over most of the EGFP emission spectrum and results in a greater than 50% signal increase at the same exposure settings compared to high-quality multiband pass filter sets.
Janson, M. , and Dogterom, M. (2006). Assembly dynamics of microtubules at molecular resolution. Nature 442, 709–712. , De Groot, C. , Gouveia, S. , Munteanu, E. , Schober, J. , Buey, R. , Hoogenraad, C. , Borisy, G. , Steinmetz, M. , et al. (2009). Mammalian end binding proteins control persistent microtubule growth. J. Cell Biol. 184, 691–706. 32 Sarah Gierke et al. , Anderson, K. , Manstein, D. , Small, J. , and Toomre, D. K. (2003). Nanometer targeting of microtubules to focal adhesions. J.
Opin. Cell Biol. 18, 179–184. Rusan, N. , Fagerstrom, C. , Yvon, A. , and Wadsworth, P. (2001). Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-alpha tubulin. Mol. Biol. Cell 12, 971–980. Salaycik, K. , Fagerstrom, C. , Tulu, U. , and Wadsworth, P. (2005). Quantification of microtubule nucleation, growth and dynamics in wound-edge cells. J. Cell Sci. 118, 4113–4122. Sammak, P. , and Borisy, G. G. (1988). Direct observation of microtubule dynamics in living cells.
Microtubules: in vivo by Lynne Cassimeris, Phong Tran