Interferometric scattering microscopy for the study of molecular motors

Book section

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 20 nm 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.

Authors: Andrecka, J; Takagi, Y; Mickolajczyk, K; Lippert, L; Sellers, J

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Elsevier
• Host title: Methods in Enzymology
• Volume: 581
• Pages: 517-539
• Series: Methods in Enzymology
• Publication date: 2016-10-10
• DOI: 10.1016/bs.mie.2016.08.016
• EISSN: 1557-7988
• ISSN: 0076-6879
• ISBN: 9780128092675
• Language: English
• Keywords: high speed; molecular motors; kinesin; myosin; single-particle tracking; dynein; interferometric scattering microscopy; single molecule