Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility

Endesfelder U, van de Linde S, Wolter S, Sauer M, Heilemann M (2010)
CHEMPHYSCHEM 11(4): 836-840.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
Subdiffraction-resolution imaging by subsequent localization of single photoswitchable molecules can achieve a spatial resolution in the range of similar to 20 nm with moderate excitation intensities, but have so far been too slow for imaging faster dynamics in biology. Herein, we introduce a novel approach for video-like subdiffraction microscopy based on rapid and reversible photoswitching of commercially available organic carbocyanine fluorophores. With the present concept, we demonstrate in vitro studies on the motility of fluorophore-labeled actin filaments along myosin II. Actin filaments were densely labeled with carbocyanine fluorophores, and the gliding velocity adjusted by the concentration of ATP. At imaging frame rates of similar to 100 Hz, only 100 consecutive frames are sufficient to generate a single high-resolution image of moving actin filaments with a lateral resolution of similar to 30 nm. A video-like sequence is generated from individual reconstructed images by additionally applying a sliding window algorithm. We measured velocities of individual actin filaments of up to similar to 0.18 mu m s(-1), observed strong bending and disruption of filaments as well as locally immobile fragments.
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Zeitschriftentitel
CHEMPHYSCHEM
Band
11
Zeitschriftennummer
4
Seite
836-840
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Endesfelder U, van de Linde S, Wolter S, Sauer M, Heilemann M. Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility. CHEMPHYSCHEM. 2010;11(4):836-840.
Endesfelder, U., van de Linde, S., Wolter, S., Sauer, M., & Heilemann, M. (2010). Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility. CHEMPHYSCHEM, 11(4), 836-840. doi:10.1002/cphc.200900944
Endesfelder, U., van de Linde, S., Wolter, S., Sauer, M., and Heilemann, M. (2010). Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility. CHEMPHYSCHEM 11, 836-840.
Endesfelder, U., et al., 2010. Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility. CHEMPHYSCHEM, 11(4), p 836-840.
U. Endesfelder, et al., “Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility”, CHEMPHYSCHEM, vol. 11, 2010, pp. 836-840.
Endesfelder, U., van de Linde, S., Wolter, S., Sauer, M., Heilemann, M.: Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility. CHEMPHYSCHEM. 11, 836-840 (2010).
Endesfelder, Ulrike, van de Linde, Sebastian, Wolter, Steve, Sauer, Markus, and Heilemann, Mike. “Subdiffraction-Resolution Fluorescence Microscopy of Myosin-Actin Motility”. CHEMPHYSCHEM 11.4 (2010): 836-840.

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