By Matthieu Piel and Manuel Théry (Eds.)
This new quantity of tools in mobilephone Biology is the second one quantity describing micropatterning, complementing quantity 119. Chapters are written via specialists within the box and comprise state of the art material.
- Includes sections on micropatterning in second with photomask, maskless micropatterning and 2nd nanopatterning
- Chapters are written by means of specialists within the field
- Cutting-edge material
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Extra resources for Micropatterning in Cell Biology Part B
Wilson, C. , Tsuchida, M. , Allen, G. , Barnhart, E. , Applegate, K. , Yam, P. , et al. (2010). Myosin II contributes to cell-scale actin network treadmilling through network disassembly. Nature, 465, 373–377. , Babcock, H. , & Zhuang, X. (2012). Dual-objective STORM reveals threedimensional filament organization in the actin cytoskeleton. Nature Methods, 9, 185–188. , & Svitkina, T. (2007). Novel roles of formin mDia2 in lamellipodia and filopodia formation in motile cells. PLoS Biology, 5, 317.
1) 4. 4 Actin dynamics in the presence of myosin freely available in solution The next step in understanding how myosins are selectively directed to specific actin substructures, while being available in the entire volume of cells, is to reconstitute actin networks driving selective contraction and disassembly induced by free myosin present in solution at the onset of actin polymerization on micropatterns. Therefore to assess how actin network architecture may control spatial and temporal recruitment of interacting motors, myosin is added directly to the polymerization mix in the presence of ATP regeneration system.
3 Structured Network Assembly photomask during the first round of UV irradiation. For actin filaments to reach and be recruited by grafted myosins, the myosin-coated micropatterns should be 20–50 mm apart from the nucleation ones. Therefore, for the second round of exposure to UV, placing the coverslip in its former position allows the microscopic shift required between the two sets of micropatterns. 2 Myosin grafting 1. Materials • Myosin, eventually GFP-labeled, should be diluted at the last minute in M-buffer to the desired concentration and stored concentrated at À20 C in 50% glycerol.