Micropatterning microtubules.
The following protocol describes a method to control the orientation and polarity of polymerizing microtubules (MTs). Reconstitution of specific geometries of dynamic MT networks is achieved using a ultraviolet (UV) micropatterning technique in combination with stabilized MT microseeds. The process is described in three main parts. First, the surface is passivated to avoid the non-specific absorption of proteins, using different polyethylene glycol (PEG)-based surface treatment. Second, specific adhesive surfaces (the micropatterns) are imprinted through a photomask using deep UVs. Lastly, MT microseeds are adhered to the micropatterns followed by MT polymerization.
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Didier
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Portran
D000818:Animals; D001360:Avidin / Q000378:metabolism; D002417:Cattle; D008856:Microscopy, Fluorescence; D055616:Microtechnology / Q000379:methods*; D008870:Microtubules / Q000378:metabolism* / Q000528:radiation effects; D011092:Polyethylene Glycols / Q000737:chemistry; D058105:Polymerization / Q000528:radiation effects; D012821:Silanes / Q000737:chemistry; D014404:Tubulin / Q000378:metabolism; D014466:Ultraviolet Rays
D012821:Silanes; D014404:Tubulin; C093483:neutravidin; D001360:Avidin; D011092:Polyethylene Glycols
Asters; Micropatterning; Microtubule antiparallel organization; Microtubule microseeds; Microtubule parallel organization; Microtubules; PLL-PEG; Silane-PEG; Surface passivation