Micropatterning Microtubules

• Published in: Methods in Cell Biology Vol. 120; pp. 39 - 51
• Main Author: Portran, Didier
• Format: Book Chapter Journal Article
• Language: English
• Published: United States Elsevier Science & Technology 2014
• Subjects: Animals; Asters; Avidin - metabolism; Cattle; Micropatterning; Microscopy, Fluorescence; Microtechnology - methods; Microtubule antiparallel organization; Microtubule microseeds; Microtubule parallel organization; Microtubules; Microtubules - metabolism; Microtubules - radiation effects; PLL-PEG; Polyethylene Glycols - chemistry; Polymerization - radiation effects; Silane-PEG; Silanes - chemistry; Surface passivation; Tubulin - metabolism; Ultraviolet Rays; Microtubule microseeds; Asters; Silane-PEG; PLL-PEG; Micropatterning; Microtubules; Microtubule parallel organization; Microtubule antiparallel organization; Surface passivation
• ISBN: 9780124171367; 0124171362
• ISSN: 0091-679X
• DOI: 10.1016/B978-0-12-417136-7.00003-3

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.