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Updated: Jan 22, 2026

Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers
Published on: May 4, 2022
Quantitative high-precision imaging of myosin-dependent filamentous actin dynamics
Sawako Yamashiro1,2, Naoki Watanabe3,4
1Laboratory of Single-Molecule Cell Biology, Kyoto University Graduate School of Biostudies, Yoshida Konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan. yamashiro.sawako.5c@kyoto-u.ac.jp.
Abstract:
Over recent decades, considerable effort has been made to understand how mechanical stress applied to the actin network alters actin assembly and disassembly dynamics. However, there are conflicting reports concerning the issue both in vitro and in cells. In this review, we discuss concerns regarding previous quantitative live-cell experiments that have attempted to evaluate myosin regulation of filamentous actin (F-actin) turnover. In particular, we highlight an error-generating mechanism in quantitative live-cell imaging, namely convection-induced misdistribution of actin-binding probes. Direct observation of actin turnover at the single-molecule level using our improved electroporation-based Single-Molecule Speckle (eSiMS) microscopy technique overcomes these concerns. We introduce our recent single-molecule analysis that unambiguously demonstrates myosin-dependent regulation of F-actin stability in live cells. We also discuss the possible application of eSiMS microscopy in the analysis of actin remodeling in striated muscle cells.
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