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.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
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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.
studies; single-molecule; superresolution fluorescence microscopy; actin-myosin motility; biophysics; photophysics
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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.

15 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Lens-based fluorescence nanoscopy.
Eggeling C, Willig KI, Sahl SJ, Hell SW., Q Rev Biophys 48(2), 2015
PMID: 25998828
Eight years of single-molecule localization microscopy.
Klein T, Proppert S, Sauer M., Histochem Cell Biol 141(6), 2014
PMID: 24496595
A simple method to estimate the average localization precision of a single-molecule localization microscopy experiment.
Endesfelder U, Malkusch S, Fricke F, Heilemann M., Histochem Cell Biol 141(6), 2014
PMID: 24522395
STED microscopy of living cells--new frontiers in membrane and neurobiology.
Eggeling C, Willig KI, Barrantes FJ., J Neurochem 126(2), 2013
PMID: 23506404
Photoactivatable synthetic fluorophores.
Raymo FM., Phys Chem Chem Phys 15(36), 2013
PMID: 23780303
Live-cell super-resolution imaging with synthetic fluorophores.
van de Linde S, Heilemann M, Sauer M., Annu Rev Phys Chem 63(), 2012
PMID: 22404589
Drift estimation for single marker switching based imaging schemes.
Geisler C, Hotz T, Schönle A, Hell SW, Munk A, Egner A., Opt Express 20(7), 2012
PMID: 22453409
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
Chemically induced photoswitching of fluorescent probes--a general concept for super-resolution microscopy.
Endesfelder U, Malkusch S, Flottmann B, Mondry J, Liguzinski P, Verveer PJ, Heilemann M., Molecules 16(4), 2011
PMID: 21490558
Direct stochastic optical reconstruction microscopy with standard fluorescent probes.
van de Linde S, Löschberger A, Klein T, Heidbreder M, Wolter S, Heilemann M, Sauer M., Nat Protoc 6(7), 2011
PMID: 21720313
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
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, Tampé R, Choquet D, Cognet L., Biophys J 99(4), 2010
PMID: 20713016
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
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
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, Fölling J, Jakobs S, Schönle A, Hell SW, Eggeling C., Biophys J 99(8), 2010
PMID: 20959110

29 References

Daten bereitgestellt von Europe PubMed Central.

Abbe, Arch. Mikr. Anat. 9(), 1873
Far-field optical nanoscopy.
Hell SW., Science 316(5828), 2007
PMID: 17525330
Advances in the speed and resolution of light microscopy.
Ji N, Shroff H, Zhong H, Betzig E., Curr. Opin. Neurobiol. 18(6), 2008
PMID: 19375302
Microscopy and its focal switch.
Hell SW., Nat. Methods 6(1), 2009
PMID: 19116611
Super-resolution fluorescence microscopy.
Huang B, Bates M, Zhuang X., Annu. Rev. Biochem. 78(), 2009
PMID: 19489737
Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission.
Klar TA, Jakobs S, Dyba M, Egner A, Hell SW., Proc. Natl. Acad. Sci. U.S.A. 97(15), 2000
PMID: 10899992
Ultra-high resolution imaging by fluorescence photoactivation localization microscopy.
Hess ST, Girirajan TP, Mason MD., Biophys. J. 91(11), 2006
PMID: 16980368
Imaging intracellular fluorescent proteins at nanometer resolution.
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF., Science 313(5793), 2006
PMID: 16902090
Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).
Rust MJ, Bates M, Zhuang X., Nat. Methods 3(10), 2006
PMID: 16896339
Subdiffraction-resolution fluorescence imaging with conventional fluorescent probes.
Heilemann M, van de Linde S, Schuttpelz M, Kasper R, Seefeldt B, Mukherjee A, Tinnefeld P, Sauer M., Angew. Chem. Int. Ed. Engl. 47(33), 2008
PMID: 18646237

AUTHOR UNKNOWN, Angew. Chem. 120(), 2008
Super-resolution imaging with small organic fluorophores.
Heilemann M, van de Linde S, Mukherjee A, Sauer M., Angew. Chem. Int. Ed. Engl. 48(37), 2009
PMID: 19670280

AUTHOR UNKNOWN, Angew. Chem. 121(), 2009
Controlling the fluorescence of ordinary oxazine dyes for single-molecule switching and superresolution microscopy.
Vogelsang J, Cordes T, Forthmann C, Steinhauer C, Tinnefeld P., Proc. Natl. Acad. Sci. U.S.A. 106(20), 2009
PMID: 19433792
Super-resolution imaging of DNA labelled with intercalating dyes.
Flors C, Ravarani CN, Dryden DT., Chemphyschem 10(13), 2009
PMID: 19554598
Fluorescence nanoscopy by ground-state depletion and single-molecule return.
Folling J, Bossi M, Bock H, Medda R, Wurm CA, Hein B, Jakobs S, Eggeling C, Hell SW., Nat. Methods 5(11), 2008
PMID: 18794861
Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics.
Shroff H, Galbraith CG, Galbraith JA, Betzig E., Nat. Methods 5(5), 2008
PMID: 18408726
Video-rate far-field optical nanoscopy dissects synaptic vesicle movement.
Westphal V, Rizzoli SO, Lauterbach MA, Kamin D, Jahn R, Hell SW., Science 320(5873), 2008
PMID: 18292304
Super-resolution video microscopy of live cells by structured illumination.
Kner P, Chhun BB, Griffis ER, Winoto L, Gustafsson MG., Nat. Methods 6(5), 2009
PMID: 19404253

Wolter, Journal of Microscopy (), 2010
Superresolution microscopy on the basis of engineered dark states.
Steinhauer C, Forthmann C, Vogelsang J, Tinnefeld P., J. Am. Chem. Soc. 130(50), 2008
PMID: 19053449

Linde, Appl. Phys. B 93(), 2008
Carbocyanine dyes as efficient reversible single-molecule optical switch.
Heilemann M, Margeat E, Kasper R, Sauer M, Tinnefeld P., J. Am. Chem. Soc. 127(11), 2005
PMID: 15771514
Assays for actin sliding movement over myosin-coated surfaces.
Kron SJ, Toyoshima YY, Uyeda TQ, Spudich JA., Meth. Enzymol. 196(), 1991
PMID: 2034132
Fluorescent actin filaments move on myosin fixed to a glass surface.
Kron SJ, Spudich JA., Proc. Natl. Acad. Sci. U.S.A. 83(17), 1986
PMID: 3462694
Evaluating atomic models of F-actin with an undecagold-tagged phalloidin derivative.
Steinmetz MO, Stoffler D, Muller SA, Jahn W, Wolpensinger B, Goldie KN, Engel A, Faulstich H, Aebi U., J. Mol. Biol. 276(1), 1998
PMID: 9514733

Shannon, Proc. IRE 37(), 1949


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