title: Imaging single molecular motor motility with total internal reflection fluorescence microscopy (TIRFM).
abstract: Total internal reflection fluorescence microscopy (TIRFM) allows fluorescent molecules on or near the plasma membrane to be visualized with a very high signal-to-noise ratio. This strategy has been very successful for imaging molecular machines as they move and do work. We provide here a general protocol for imaging single molecular motors as they move along microtubule tracks. Our protocol is designed for the study of cytoplasmic dynein purified from Saccharomyces cerevisiae, but it represents a general framework for any in vitro single-molecule assay.
authors.forename: Samara L
authors.initials: SL
authors.lastname: Reck-Peterson
authors.forename: Nathan D
authors.initials: ND
authors.lastname: Derr
authors.forename: Nico
authors.initials: N
authors.lastname: Stuurman
mesh_terms: D002462:Cell Membrane / Q000737:chemistry*; D003593:Cytoplasm / Q000737:chemistry*; D004398:Dyneins / Q000032:analysis; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D020409:Molecular Motor Proteins / Q000032:analysis*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis
chemical_list: D020409:Molecular Motor Proteins; D029701:Saccharomyces cerevisiae Proteins; D004398:Dyneins