关键词: Dynein High speed Interferometric scattering microscopy Kinesin Molecular motors Myosin Single molecule Single-particle tracking

Mesh : Dyneins / chemistry isolation & purification Humans Kinesins / chemistry isolation & purification Microscopy, Fluorescence / methods Microscopy, Interference / methods Molecular Motor Proteins / chemistry metabolism Myosins / chemistry isolation & purification

来  源:   DOI:10.1016/bs.mie.2016.08.016

Abstract:
Our understanding of molecular motor function has been greatly improved by the development of imaging modalities, which enable real-time observation of their motion at the single-molecule level. Here, we describe the use of a new method, interferometric scattering microscopy, for the investigation of motor protein dynamics by attaching and tracking the motion of metallic nanoparticle labels as small as 20nm diameter. Using myosin-5, kinesin-1, and dynein as examples, we describe the basic assays, labeling strategies, and principles of data analysis. Our approach is relevant not only for motor protein dynamics but also provides a general tool for single-particle tracking with high spatiotemporal precision, which overcomes the limitations of single-molecule fluorescence methods.
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