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Simultaneous multi-lifetime multi-color STED imaging for colocalization analysesJohanna Bückers, Dominik Wildanger, Giuseppe Vicidomini, Lars Kastrup, and Stefan W. Hell »View Author Affiliations
Johanna Bückers,
Dominik Wildanger,
Giuseppe Vicidomini,
Lars Kastrup,
and Stefan W. Hell*
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany *Corresponding author: shell@gwdg.de |
Optics Express, Vol. 19, Issue 4, pp. 3130-3143 (2011)
http://dx.doi.org/10.1364/OE.19.003130
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Abstract
We describe a STED microscope optimized for colocalization experiments with up to three colors. Two fluorescence labels are separated by their fluorescence lifetime whereas a third channel is discriminated by the wavelength of fluorescence emission. Since it does not require a second STED beam, separating by lifetime is insensitive to drift and thus optimally suited for colocalization analyses. Furthermore, we propose a setup having a second STED beam for long duration multicolor recording.
© 2011 OSA
OCIS Codes
(180.2520) Microscopy : Fluorescence microscopy
(350.5730) Other areas of optics : Resolution
ToC Category:
Microscopy
History
Original Manuscript: January 4, 2011
Manuscript Accepted: January 18, 2011
Published: February 2, 2011
Virtual Issues
Vol. 6, Iss. 3 Virtual Journal for Biomedical Optics
Citation
Johanna Bückers, Dominik Wildanger, Giuseppe Vicidomini, Lars Kastrup, and Stefan W. Hell, "Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses," Opt. Express 19, 3130-3143 (2011)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-19-4-3130
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References
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- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
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- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
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- M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “An experimental comparison of the maximum likelihood estimation and nonlinear least-squares fluorescence lifetime analysis of single molecules,” Anal. Chem. 73(9), 2078–2086 (2001). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
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- 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(5793), 1642–1645 (2006). [CrossRef] [PubMed]
- M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “An experimental comparison of the maximum likelihood estimation and nonlinear least-squares fluorescence lifetime analysis of single molecules,” Anal. Chem. 73(9), 2078–2086 (2001). [CrossRef] [PubMed]
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- M. Bates, B. Huang, G. T. Dempsey, and X. W. Zhuang, “Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes,” Science 317(5845), 1749–1753 (2007). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- T. Foerster, “Intermolecular energy migration and fluorescence,” Ann. Phys. 2, 55 (1948).
- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
- H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig, “Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes,” Proc. Natl. Acad. Sci. U.S.A. 104(51), 20308–20313 (2007). [CrossRef] [PubMed]
- H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig, “Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes,” Proc. Natl. Acad. Sci. U.S.A. 104(51), 20308–20313 (2007). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “An experimental comparison of the maximum likelihood estimation and nonlinear least-squares fluorescence lifetime analysis of single molecules,” Anal. Chem. 73(9), 2078–2086 (2001). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
- H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig, “Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes,” Proc. Natl. Acad. Sci. U.S.A. 104(51), 20308–20313 (2007). [CrossRef] [PubMed]
- S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization microscopy,” Biophys. J. 91(11), 4258–4272 (2006). [CrossRef] [PubMed]
- S. van de Linde, R. Kasper, M. Heilemann, and M. Sauer, “Photoswitching microscopy with standard fluorophores,” Appl. Phys. B 93(4), 725–731 (2008). [CrossRef]
- D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, and S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 1–4 (2010). [CrossRef]
- D. Wildanger, R. Medda, L. Kastrup, and S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009). [CrossRef] [PubMed]
- S. W. Hell, “Microscopy and its focal switch,” Nat. Methods 6(1), 24–32 (2009). [CrossRef] [PubMed]
- V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell, “Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement,” Science 320(5873), 246–249 (2008). [CrossRef] [PubMed]
- D. Wildanger, E. Rittweger, L. Kastrup, and S. W. Hell, “STED microscopy with a supercontinuum laser source,” Opt. Express 16(13), 9614–9621 (2008). [CrossRef] [PubMed]
- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- J. Keller, A. Schönle, and S. W. Hell, “Efficient fluorescence inhibition patterns for RESOLFT microscopy,” Opt. Express 15(6), 3361–3371 (2007). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- K. I. Willig, S. O. Rizzoli, V. Westphal, R. Jahn, and S. W. Hell, “STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis,” Nature 440(7086), 935–939 (2006). [CrossRef] [PubMed]
- S. W. Hell and J. Wichmann, “Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy,” Opt. Lett. 19(11), 780–782 (1994). [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(5793), 1642–1645 (2006). [CrossRef] [PubMed]
- S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization microscopy,” Biophys. J. 91(11), 4258–4272 (2006). [CrossRef] [PubMed]
- M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “An experimental comparison of the maximum likelihood estimation and nonlinear least-squares fluorescence lifetime analysis of single molecules,” Anal. Chem. 73(9), 2078–2086 (2001). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
- C. D. Hu, Y. Chinenov, and T. K. Kerppola, “Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation,” Mol. Cell 9(4), 789–798 (2002). [CrossRef] [PubMed]
- M. Bates, B. Huang, G. T. Dempsey, and X. W. Zhuang, “Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes,” Science 317(5845), 1749–1753 (2007). [CrossRef] [PubMed]
- V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell, “Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement,” Science 320(5873), 246–249 (2008). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- K. I. Willig, S. O. Rizzoli, V. Westphal, R. Jahn, and S. W. Hell, “STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis,” Nature 440(7086), 935–939 (2006). [CrossRef] [PubMed]
- D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, and S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 1–4 (2010). [CrossRef]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell, “Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement,” Science 320(5873), 246–249 (2008). [CrossRef] [PubMed]
- S. van de Linde, R. Kasper, M. Heilemann, and M. Sauer, “Photoswitching microscopy with standard fluorophores,” Appl. Phys. B 93(4), 725–731 (2008). [CrossRef]
- D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, and S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 1–4 (2010). [CrossRef]
- D. Wildanger, R. Medda, L. Kastrup, and S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009). [CrossRef] [PubMed]
- D. Wildanger, E. Rittweger, L. Kastrup, and S. W. Hell, “STED microscopy with a supercontinuum laser source,” Opt. Express 16(13), 9614–9621 (2008). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- J. Keller, A. Schönle, and S. W. Hell, “Efficient fluorescence inhibition patterns for RESOLFT microscopy,” Opt. Express 15(6), 3361–3371 (2007). [CrossRef] [PubMed]
- T. K. Kerppola, “Design and implementation of bimolecular fluorescence complementation (BiFC) assays for the visualization of protein interactions in living cells,” Nat. Protoc. 1(3), 1278–1286 (2006). [CrossRef]
- C. D. Hu, Y. Chinenov, and T. K. Kerppola, “Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation,” Mol. Cell 9(4), 789–798 (2002). [CrossRef] [PubMed]
- R. E. Thompson, D. R. Larson, and W. W. Webb, “Precise nanometer localization analysis for individual fluorescent probes,” Biophys. J. 82(5), 2775–2783 (2002). [CrossRef] [PubMed]
- V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell, “Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement,” Science 320(5873), 246–249 (2008). [CrossRef] [PubMed]
- K. Carlsson and A. Liljeborg, “Confocal fluorescence microscopy using spectral and lifetime information to simultaneously record four fluorophores with high channel separation,” J. Microsc. 185(1), 37–46 (1997). [CrossRef]
- 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(5793), 1642–1645 (2006). [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(5793), 1642–1645 (2006). [CrossRef] [PubMed]
- S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization microscopy,” Biophys. J. 91(11), 4258–4272 (2006). [CrossRef] [PubMed]
- M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “An experimental comparison of the maximum likelihood estimation and nonlinear least-squares fluorescence lifetime analysis of single molecules,” Anal. Chem. 73(9), 2078–2086 (2001). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
- D. Wildanger, R. Medda, L. Kastrup, and S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009). [CrossRef] [PubMed]
- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
- R. Neher and E. Neher, “Optimizing imaging parameters for the separation of multiple labels in a fluorescence image,” J. Microsc. 213(1), 46–62 (2004). [CrossRef]
- R. Neher and E. Neher, “Optimizing imaging parameters for the separation of multiple labels in a fluorescence image,” J. Microsc. 213(1), 46–62 (2004). [CrossRef]
- D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, and S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 1–4 (2010). [CrossRef]
- V. Zinchuk, O. Zinchuk, and T. Okada, “Quantitative Colocalization Analysis of Multicolor Confocal Immunofluorescence Microscopy Images: Pushing Pixels to Explore Biological Phenomena,” Acta Histochem. Cytochem. 40(4), 101–111 (2007). [CrossRef] [PubMed]
- H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig, “Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes,” Proc. Natl. Acad. Sci. U.S.A. 104(51), 20308–20313 (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(5793), 1642–1645 (2006). [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(5793), 1642–1645 (2006). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
- V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell, “Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement,” Science 320(5873), 246–249 (2008). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- K. I. Willig, S. O. Rizzoli, V. Westphal, R. Jahn, and S. W. Hell, “STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis,” Nature 440(7086), 935–939 (2006). [CrossRef] [PubMed]
- M. J. Rust, M. Bates, and X. W. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
- S. van de Linde, R. Kasper, M. Heilemann, and M. Sauer, “Photoswitching microscopy with standard fluorophores,” Appl. Phys. B 93(4), 725–731 (2008). [CrossRef]
- M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “An experimental comparison of the maximum likelihood estimation and nonlinear least-squares fluorescence lifetime analysis of single molecules,” Anal. Chem. 73(9), 2078–2086 (2001). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- J. Keller, A. Schönle, and S. W. Hell, “Efficient fluorescence inhibition patterns for RESOLFT microscopy,” Opt. Express 15(6), 3361–3371 (2007). [CrossRef] [PubMed]
- M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “An experimental comparison of the maximum likelihood estimation and nonlinear least-squares fluorescence lifetime analysis of single molecules,” Anal. Chem. 73(9), 2078–2086 (2001). [CrossRef] [PubMed]
- K. Carlsson, N. Aslund, K. Mossberg, J. Philip, M. Maus, M. Cotlet, J. Hofkens, T. Gensch, F. C. De Schryver, J. Schaffer, and C. A. Seidel, “Simultaneous confocal recording of multiple fluorescent labels with improved channel separation,” J. Microsc. 176(Pt 3), 287–299 (1994). [CrossRef] [PubMed]
- H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig, “Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes,” Proc. Natl. Acad. Sci. U.S.A. 104(51), 20308–20313 (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(5793), 1642–1645 (2006). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- R. E. Thompson, D. R. Larson, and W. W. Webb, “Precise nanometer localization analysis for individual fluorescent probes,” Biophys. J. 82(5), 2775–2783 (2002). [CrossRef] [PubMed]
- S. van de Linde, R. Kasper, M. Heilemann, and M. Sauer, “Photoswitching microscopy with standard fluorophores,” Appl. Phys. B 93(4), 725–731 (2008). [CrossRef]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- R. E. Thompson, D. R. Larson, and W. W. Webb, “Precise nanometer localization analysis for individual fluorescent probes,” Biophys. J. 82(5), 2775–2783 (2002). [CrossRef] [PubMed]
- A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A. C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, “Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters,” Biophys. J. 93(9), 3285–3290 (2007). [CrossRef] [PubMed]
- V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell, “Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement,” Science 320(5873), 246–249 (2008). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
- K. I. Willig, S. O. Rizzoli, V. Westphal, R. Jahn, and S. W. Hell, “STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis,” Nature 440(7086), 935–939 (2006). [CrossRef] [PubMed]
- H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig, “Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes,” Proc. Natl. Acad. Sci. U.S.A. 104(51), 20308–20313 (2007). [CrossRef] [PubMed]
- D. Wildanger, R. Medda, L. Kastrup, and S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009). [CrossRef] [PubMed]
- D. Wildanger, E. Rittweger, L. Kastrup, and S. W. Hell, “STED microscopy with a supercontinuum laser source,” Opt. Express 16(13), 9614–9621 (2008). [CrossRef] [PubMed]
- K. I. Willig, S. O. Rizzoli, V. Westphal, R. Jahn, and S. W. Hell, “STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis,” Nature 440(7086), 935–939 (2006). [CrossRef] [PubMed]
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- M. Bates, B. Huang, G. T. Dempsey, and X. W. Zhuang, “Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes,” Science 317(5845), 1749–1753 (2007). [CrossRef] [PubMed]
- M. J. Rust, M. Bates, and X. W. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
- V. Zinchuk, O. Zinchuk, and T. Okada, “Quantitative Colocalization Analysis of Multicolor Confocal Immunofluorescence Microscopy Images: Pushing Pixels to Explore Biological Phenomena,” Acta Histochem. Cytochem. 40(4), 101–111 (2007). [CrossRef] [PubMed]
- V. Zinchuk, O. Zinchuk, and T. Okada, “Quantitative Colocalization Analysis of Multicolor Confocal Immunofluorescence Microscopy Images: Pushing Pixels to Explore Biological Phenomena,” Acta Histochem. Cytochem. 40(4), 101–111 (2007). [CrossRef] [PubMed]
Acta Histochem. Cytochem.
- V. Zinchuk, O. Zinchuk, and T. Okada, “Quantitative Colocalization Analysis of Multicolor Confocal Immunofluorescence Microscopy Images: Pushing Pixels to Explore Biological Phenomena,” Acta Histochem. Cytochem. 40(4), 101–111 (2007). [CrossRef] [PubMed]
Anal. Biochem.
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Anal. Chem.
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Ann. Phys.
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Appl. Phys. B
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Biophys. J.
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- S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization microscopy,” Biophys. J. 91(11), 4258–4272 (2006). [CrossRef] [PubMed]
- R. E. Thompson, D. R. Larson, and W. W. Webb, “Precise nanometer localization analysis for individual fluorescent probes,” Biophys. J. 82(5), 2775–2783 (2002). [CrossRef] [PubMed]
- G. Donnert, J. Keller, C. A. Wurm, S. O. Rizzoli, V. Westphal, A. Schönle, R. Jahn, S. Jakobs, C. Eggeling, and S. W. Hell, “Two-Color Far-Field Fluorescence Nanoscopy,” Biophys. J. 92(8), L67–L69 (2007). [CrossRef] [PubMed]
J. Microsc.
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Mol. Cell
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Nano Lett.
- M. Bossi, J. Folling, V. N. Belov, V. P. Boyarskiy, R. Medda, A. Egner, C. Eggeling, A. Schonle, and S. W. Hell, “Multi-color far-field fluorescence nanoscopy through isolated detection of distinct molecular species,” Nano Lett. 8(8), 2463–2468 (2008). [CrossRef] [PubMed]
Nat. Methods
- S. W. Hell, “Microscopy and its focal switch,” Nat. Methods 6(1), 24–32 (2009). [CrossRef] [PubMed]
- M. J. Rust, M. Bates, and X. W. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
Nat. Protoc.
- T. K. Kerppola, “Design and implementation of bimolecular fluorescence complementation (BiFC) assays for the visualization of protein interactions in living cells,” Nat. Protoc. 1(3), 1278–1286 (2006). [CrossRef]
Nature
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Opt. Express
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Opt. Lett.
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PMC Biophys
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Proc. Natl. Acad. Sci. U.S.A.
- H. Shroff, C. G. Galbraith, J. A. Galbraith, H. White, J. Gillette, S. Olenych, M. W. Davidson, and E. Betzig, “Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes,” Proc. Natl. Acad. Sci. U.S.A. 104(51), 20308–20313 (2007). [CrossRef] [PubMed]
Science
- V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell, “Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement,” Science 320(5873), 246–249 (2008). [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(5793), 1642–1645 (2006). [CrossRef] [PubMed]
- M. Bates, B. Huang, G. T. Dempsey, and X. W. Zhuang, “Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes,” Science 317(5845), 1749–1753 (2007). [CrossRef] [PubMed]
Other
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2010, Neumann, PMC Biophys
- D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, and S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 1–4 (2010). [CrossRef]
- D. Wildanger, R. Medda, L. Kastrup, and S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009). [CrossRef] [PubMed]
- S. W. Hell, “Microscopy and its focal switch,” Nat. Methods 6(1), 24–32 (2009). [CrossRef] [PubMed]
- S. van de Linde, R. Kasper, M. Heilemann, and M. Sauer, “Photoswitching microscopy with standard fluorophores,” Appl. Phys. B 93(4), 725–731 (2008). [CrossRef]
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