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Updated: Jun 15, 2026

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Myosin-Specific Adaptations of In vitro Fluorescence Microscopy-Based Motility Assays

Published on: February 4, 2021

Subdiffraction-resolution fluorescence microscopy of myosin-actin motility.

Ulrike Endesfelder¹, Sebastian van de Linde, Steve Wolter

¹Applied Laser Physics and Laser Spectroscopy, Physics Department, Bielefeld University, Universitätsstrasse 25, 33615 Bielefeld, Germany.

Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry

|February 27, 2010

Summary

This study introduces rapid photoswitching microscopy for high-resolution imaging of biological dynamics. The novel method achieves ~30 nm resolution, enabling visualization of fast-moving actin filaments at ~100 Hz.

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Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

Published on: December 1, 2016

Area of Science:

Biophysics
Microscopy
Molecular Biology

Background:

Subdiffraction-resolution microscopy offers high spatial detail but is often too slow for dynamic biological processes.
Existing methods struggle to capture rapid molecular movements due to slow imaging speeds.

Purpose of the Study:

To develop a video-like subdiffraction microscopy technique for imaging fast biological dynamics.
To enable high-resolution visualization of motile biological structures using rapid photoswitching.

Main Methods:

Utilized rapid and reversible photoswitching of carbocyanine fluorophores for microscopy.
Applied a sliding window algorithm to reconstruct video-like sequences from ~100 Hz frame rates.
In vitro studies on fluorophore-labeled actin filament motility along myosin II, with velocity controlled by ATP concentration.

Main Results:

Achieved a lateral resolution of approximately 30 nm for moving actin filaments.
Captured individual actin filament velocities up to approximately 0.18 micrometers per second.
Observed filament bending, disruption, and immobile fragments during motility assays.

Conclusions:

The developed rapid photoswitching microscopy enables high-resolution, video-like imaging of fast biological dynamics.
This technique provides new insights into the mechanics and behavior of motile molecular systems.
The method is suitable for studying complex dynamic processes in biophysics and molecular biology.