Tracking Single Kinesin in Live Cells Using MINFLUX

Takahiro Deguchi; Nikolay Arkadievich Sergeev; Jonas Ries

doi:10.1007/978-1-0716-4280-1_5

Tracking Single Kinesin in Live Cells Using MINFLUX

Methods Mol Biol. 2025:2881:119-131. doi: 10.1007/978-1-0716-4280-1_5.

Authors

Takahiro Deguchi¹, Nikolay Arkadievich Sergeev^{1

2

3}, Jonas Ries^{4

5

6

7}

Affiliations

¹ European Molecular Biology Laboratory, Cell Biology and Biophysics, Heidelberg, Germany.
² Max Perutz Labs, Vienna Biocenter Campus, Vienna, Austria.
³ Department of Structural and Computational Biology, Center for Molecular Biology, University of Vienna, Vienna, Austria.
⁴ European Molecular Biology Laboratory, Cell Biology and Biophysics, Heidelberg, Germany. jonas.ries@univie.ac.at.
⁵ Max Perutz Labs, Vienna Biocenter Campus, Vienna, Austria. jonas.ries@univie.ac.at.
⁶ Department of Structural and Computational Biology, Center for Molecular Biology, University of Vienna, Vienna, Austria. jonas.ries@univie.ac.at.
⁷ Faculty of Physics, University of Vienna, Vienna, Austria. jonas.ries@univie.ac.at.

PMID: 39704940
DOI: 10.1007/978-1-0716-4280-1_5

Abstract

MINFLUX is a super-resolution fluorescence microscopy technique that enables single-molecule tracking in live cells at a single-nanometer spatial and sub-millisecond temporal resolution. This chapter describes a method for tracking fluorescently labeled human kinesin-1 in live cells using MINFLUX and analyzing kinesin stepping dynamics.

Keywords: Conformational dynamics; Fluorescence microscopy; Kinesin; MINFLUX; Motor protein; Single-molecule tracking; Super-resolution.

MeSH terms

Humans
Kinesins* / metabolism
Microscopy, Fluorescence* / methods
Microtubules / metabolism
Single Molecule Imaging* / methods

Substances

Kinesins