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.

Journal Article | Published | English

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Abstract
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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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.
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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Data provided by Europe PubMed Central.

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PMID: 25998828
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Endesfelder U, Malkusch S, Fricke F, Heilemann M., Histochem. Cell Biol. 141(6), 2014
PMID: 24522395
Eight years of single-molecule localization microscopy.
Klein T, Proppert S, Sauer M., Histochem. Cell Biol. 141(6), 2014
PMID: 24496595
Photoactivatable synthetic fluorophores.
Raymo FM., Phys Chem Chem Phys 15(36), 2013
PMID: 23780303
STED microscopy of living cells--new frontiers in membrane and neurobiology.
Eggeling C, Willig KI, Barrantes FJ., J. Neurochem. 126(2), 2013
PMID: 23506404
Drift estimation for single marker switching based imaging schemes.
Geisler C, Hotz T, Schonle A, Hell SW, Munk A, Egner A., Opt Express 20(7), 2012
PMID: 22453409
Live-cell super-resolution imaging with synthetic fluorophores.
van de Linde S, Heilemann M, Sauer M., Annu Rev Phys Chem 63(), 2012
PMID: 22404589
Direct stochastic optical reconstruction microscopy with standard fluorescent probes.
van de Linde S, Loschberger A, Klein T, Heidbreder M, Wolter S, Heilemann M, Sauer M., Nat Protoc 6(7), 2011
PMID: 21720313
Molecular strategies to read and write at the nanoscale with far-field optics.
Cusido J, Impellizzeri S, Raymo FM., Nanoscale 3(1), 2011
PMID: 20936237
Multicolor fluorescence nanoscopy in fixed and living cells by exciting conventional fluorophores with a single wavelength.
Testa I, Wurm CA, Medda R, Rothermel E, von Middendorf C, Folling J, Jakobs S, Schonle A, Hell SW, Eggeling C., Biophys. J. 99(8), 2010
PMID: 20959110
Dynamic superresolution imaging of endogenous proteins on living cells at ultra-high density.
Giannone G, Hosy E, Levet F, Constals A, Schulze K, Sobolevsky AI, Rosconi MP, Gouaux E, Tampe R, Choquet D, Cognet L., Biophys. J. 99(4), 2010
PMID: 20713016
Live-cell super-resolution imaging with trimethoprim conjugates.
Wombacher R, Heidbreder M, van de Linde S, Sheetz MP, Heilemann M, Cornish VW, Sauer M., Nat. Methods 7(9), 2010
PMID: 20693998
Surfing on a new wave of single-molecule fluorescence methods.
Hohlbein J, Gryte K, Heilemann M, Kapanidis AN., Phys Biol 7(3), 2010
PMID: 20686191
Make them blink: probes for super-resolution microscopy.
Vogelsang J, Steinhauer C, Forthmann C, Stein IH, Person-Skegro B, Cordes T, Tinnefeld P., Chemphyschem 11(12), 2010
PMID: 20632356

29 References

Data provided by Europe PubMed Central.

Super-resolution fluorescence microscopy.
Huang B, Bates M, Zhuang X., Annu. Rev. Biochem. 78(), 2009
PMID: 19489737
Super-resolution imaging of DNA labelled with intercalating dyes.
Flors C, Ravarani CN, Dryden DT., Chemphyschem 10(13), 2009
PMID: 19554598
Hochauflösende Mikroskopie mit kleinen organischen Farbstoffen
Heilemann, Angewandte Chemie 121(37), 2009
Super-Resolution Imaging with Small Organic Fluorophores
Heilemann, Angewandte Chemie International Edition 48(37), 2009

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