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Nanoscale separation of molecular species based on their rotational mobility
Ilaria Testa, Andreas Schönle, Claas v. Middendorff, Claudia Geisler, Rebecca Medda, Christian A. Wurm, Andre C. Stiel, Stefan Jakobs, Mariano Bossi, Christian Eggeling, Stefan W. Hell, and Alexander Egner »View Author Affiliations
Ilaria Testa,
Andreas Schönle,*
Claas v. Middendorff,
Claudia Geisler,
Rebecca Medda,
Christian A. Wurm,
Andre C. Stiel,
Stefan Jakobs,
Mariano Bossi,
Christian Eggeling,
Stefan W. Hell,
and Alexander Egner
Department of NanoBiophotonics, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany
*Corresponding author: aschoen@gwdg.de
Optics Express, Vol. 16, Issue 25, pp. 21093-21104 (2008)
http://dx.doi.org/10.1364/OE.16.021093
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Abstract
We combine far-field fluorescence nanoscopy through serialized recording of switchable emitters with polarization-sensitive fluorescence detection. In addition to imaging with nanoscale spatial resolution, this technique allows determination of the fluorescence anisotropy of each detected dipole emitter and thus an estimate of its rotational mobility. Sub-populations of fluorescent markers can thus be separated based on their interaction with the sample. We applied this new functional nanoscopy to imaging of living mammalian cells.
© 2008 Optical Society of America
OCIS Codes
(110.0180) Imaging systems : Microscopy
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(170.6280) Medical optics and biotechnology : Spectroscopy, fluorescence and luminescence
(180.2520) Microscopy : Fluorescence microscopy
ToC Category:
Microscopy
History
Original Manuscript: November 12, 2008
Revised Manuscript: December 3, 2008
Manuscript Accepted: December 3, 2008
Published: December 4, 2008
Virtual Issues
Vol. 4, Iss. 2 Virtual Journal for Biomedical Optics
Citation
Ilaria Testa, Andreas Schönle, Claas v. Middendorff, Claudia Geisler, Rebecca Medda, Christian A. Wurm, Andre C. Stiel, Stefan Jakobs, Mariano Bossi, Christian Eggeling, Stefan W. Hell, and Alexander Egner, "Nanoscale separation of molecular species based on their rotational mobility," Opt. Express 16, 21093-21104 (2008)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-16-25-21093
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References
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- M. Bossi, J. Fölling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schönle, and S. W. Hell, "Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species," Nano Lett. 8, 2463-2468 (2008). [CrossRef] [PubMed]
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- T. D. Lacoste, X. Michalet, F. Pinaud, D. S. Chemla, A. P. Alivisatos, and S. Weiss, "Ultrahigh-resolution multicolor colocalization of single fluorescent probes," Proc. Natl. Acad. Sci. USA 97, 9461-9466 (2000). [CrossRef] [PubMed]
- T. Ha, T. Enderle, D. F. Ogletree, D. S. Chemla, P. R. Selvin, and S. Weiss, "Probing the interaction between two single molecules: Fluorescence resonance energy transfer between a single donor and a single acceptor," Proc. Natl. Acad. Sci. USA 93, 6264-6268 (1996). [CrossRef] [PubMed]
- T. Ha, T. Enderle, D. S. Chemla, P. R. Selvin, and S. Weiss, "Single molecule dynamics studied by polarization modulation," Phys. Rev. Lett. 77, 3979-3982 (1996). [CrossRef] [PubMed]
- P. Lamesch, N. Li, S. Milstein, C. Fan, T. Hao, G. Szabo, Z. Hu, K. Venkatesan, G. Bethel, P. Martin, J. Rogers, S. Lawlor, S. McLaren, A. Dricot, H. Borick, M. E. Cusick, J. Vandenhaute, I. Dunham, D. E. Hill, and M. Vidal, "hORFeome v3.1: A resource of human open reading frames representing over 10,000 human genes," Genomics 89, 307-315 (2007). [CrossRef] [PubMed]
- E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, "Imaging Intracellular Fluorescent Proteins at Nanometer Resolution," Science 313, 1642-1645 (2006). [CrossRef] [PubMed]
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- C. Zander, M. Sauer, K. H. Drexhage, D.-S. Ko, A. Schulz, J. Wolfrum, L. Brand, C. Eggeling, and C. A. M. Seidel, "Detection and characterization of single molecules in aqueous solution," Appl. Phys. B 63, 517-523 (1996).
- P. Lamesch, N. Li, S. Milstein, C. Fan, T. Hao, G. Szabo, Z. Hu, K. Venkatesan, G. Bethel, P. Martin, J. Rogers, S. Lawlor, S. McLaren, A. Dricot, H. Borick, M. E. Cusick, J. Vandenhaute, I. Dunham, D. E. Hill, and M. Vidal, "hORFeome v3.1: A resource of human open reading frames representing over 10,000 human genes," Genomics 89, 307-315 (2007). [CrossRef] [PubMed]
- P. Lamesch, N. Li, S. Milstein, C. Fan, T. Hao, G. Szabo, Z. Hu, K. Venkatesan, G. Bethel, P. Martin, J. Rogers, S. Lawlor, S. McLaren, A. Dricot, H. Borick, M. E. Cusick, J. Vandenhaute, I. Dunham, D. E. Hill, and M. Vidal, "hORFeome v3.1: A resource of human open reading frames representing over 10,000 human genes," Genomics 89, 307-315 (2007). [CrossRef] [PubMed]
- T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, "Fluorescence microscopy with diffraction resolution limit broken by stimulated emission," Proc. Nat. Acad. Sci. U.S.A 97, 8206-8210 (2000). [CrossRef]
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- J. Fölling, V. Belov, D. Riedel, A. Schönle, A. Egner, C. Eggeling, M. Bossi, and S. W. Hell "Fluorescence Nanoscopy with Optical Sectioning by Two-Photon Induced Molecular Switching using Continuous-Wave Lasers," ChemPhysChem 9, 321 - 326 (2008). [CrossRef] [PubMed]
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- M. Bossi, J. Fölling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schönle, and S. W. Hell, "Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species," Nano Lett. 8, 2463-2468 (2008). [CrossRef] [PubMed]
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- T. Schmidt, G. J. Schutz, W. Baumgartner, H. J. Gruber, and H. Schindler, "Imaging of single molecule diffusion," Proc. Natl. Acad. Sci. USA 93, 2926-9 (1996). [CrossRef] [PubMed]
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- E. B. Shera, N. K. Seitzinger, L. M. Davis, R. A. Keller, and S. A. Soper, "Detection of single fluorescent molecules," Chem. Phys. Lett. 174, 553-557 (1990). [CrossRef]
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Angew. Chem. Int. Ed.
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Appl. Phys. A
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Appl. Phys. B
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Biophys. J.
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Chem. Phys. Lett.
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ChemPhysChem
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Genomics
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J. Phys. Chem. A
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Langmuir
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Methods
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Nano Lett.
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Nat. Biotechnol.
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Nat. Meth.
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Nat. Methods
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Nature Biotechnol.
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Proc. Nat. Acad. Sci. U.S.A
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