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Two-color STED microscopy in living cellsPatrina A. Pellett, Xiaoli Sun, Travis J. Gould, James E. Rothman, Ming-Qun Xu, Ivan R. Corrêa, and Joerg Bewersdorf »View Author Affiliations
Patrina A. Pellett,1,2
Xiaoli Sun,3
Travis J. Gould,1
James E. Rothman,1,2
Ming-Qun Xu,3
Ivan R. Corrêa,3
and Joerg Bewersdorf1,4,*
1Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, Connecticut 06510, USA 2Department of Chemistry, Yale University, 225 Prospect Street, New Haven, Connecticut 06510, USA 3New England Biolabs, 240 County Road, Ipswich, Massachusetts 01938, USA 4Department of Biomedical Engineering, Yale University, 333 Cedar Street, New Haven, Connecticut 06510, USA *Corresponding author: joerg.bewersdorf@yale.edu |
Biomedical Optics Express, Vol. 2, Issue 8, pp. 2364-2371 (2011)
http://dx.doi.org/10.1364/BOE.2.002364
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Abstract
Diffraction-unlimited resolution provided by Stimulated Emission Depletion (STED) microscopy allows for imaging cellular processes in living cells that are not visible by conventional microscopy. However, it has so far not been possible to study dynamic nanoscale interactions because multicolor live cell STED microscopy has yet to be demonstrated and suitable labeling technologies and protocols are lacking. Here we report the first realization of two-color STED imaging in living cells. Using improved SNAPf and CLIPf technologies to label epidermal growth factor (EGF) and EGF receptor (EGFR), we report resolutions of 78 nm and 82 nm for 22 sequential two-color scans in living cells.
© 2011 OSA
OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.2520) Microscopy : Fluorescence microscopy
(350.5730) Other areas of optics : Resolution
ToC Category:
Microscopy
History
Original Manuscript: June 15, 2011
Revised Manuscript: July 21, 2011
Manuscript Accepted: July 21, 2011
Published: July 22, 2011
Citation
Patrina A. Pellett, Xiaoli Sun, Travis J. Gould, James E. Rothman, Ming-Qun Xu, Ivan R. Corrêa, and Joerg Bewersdorf, "Two-color STED microscopy in living cells," Biomed. Opt. Express 2, 2364-2371 (2011)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-2-8-2364
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References
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- 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]
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- B. Hein, K. I. Willig, C. A. Wurm, V. Westphal, S. Jakobs, and S. W. Hell, “Stimulated emission depletion nanoscopy of living cells using SNAP-tag fusion proteins,” Biophys. J. 98(1), 158–163 (2010). [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. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
- S. W. Hell, “Microscopy and its focal switch,” Nat. Methods 6(1), 24–32 (2009). [CrossRef] [PubMed]
- L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, and S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008). [CrossRef] [PubMed]
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- X. Sun, A. Zhang, B. Baker, L. Sun, A. Howard, J. Buswell, D. Maurel, A. Masharina, K. Johnsson, C. J. Noren, M.-Q. Xu, and I. R. Corrêa., “Development of SNAP-tag fluorogenic probes for wash-free fluoescence imaging,” ChemBioChem . in press. [PubMed]
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- R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan, and B. S. Glick, “A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling,” Biochemistry 48(35), 8279–8281 (2009). [CrossRef] [PubMed]
- J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, and S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011). [CrossRef] [PubMed]
- R. Schmidt, C. A. Wurm, A. Punge, A. Egner, S. Jakobs, and S. W. Hell, “Mitochondrial cristae revealed with focused light,” Nano Lett. 9(6), 2508–2510 (2009). [CrossRef] [PubMed]
- T. J. Gould, J. R. Myers, and J. Bewersdorf, “Total internal reflection STED microscopy,” Opt. Express 19(14), 13351–13357 (2011). [CrossRef] [PubMed]
Arrivoli, C.
Bachmann, J.
Baker, B.
Bates, M.
Beaufils, F.
Belov, V. N.
Berget, P. B.
Betzig, E.
Bewersdorf, J.
Bhattacharyya, D.
Bierwagen, J.
Bonifacino, J. S.
Bruchez, M. P.
Bückers, J.
Buswell, J.
Carpenter, G.
Carter, R. E.
Corrêa, I. R.
Davidson, M. W.
Donnert, G.
Dyba, M.
Eggeling, C.
Egner, A.
Ehrhard, T.
Engelhardt, J.
Fitzpatrick, J. A. J.
Gautier, A.
Gendreizig, S.
Girirajan, T. P. K.
Glick, B. S.
Gould, T. J.
Gronemeyer, T.
Harke, B.
Hein, B.
Heinis, C.
Hell, S. W.
Herten, D. P.
Hess, H. F.
Hess, S. T.
Howard, A.
Jahn, R.
Jakobs, S.
Johnsson, K.
Juillerat, A.
Kamin, D.
Kastrup, L.
Keenan, R. J.
Keller, J.
Keppler, A.
Kindermann, M.
Klingmüller, U.
Kolmakov, K.
Lauterbach, M. A.
Leutenegger, M.
Lindwasser, O. W.
Lippincott-Schwartz, J.
Lymperopoulos, K.
Masharina, A.
Mason, M. D.
Maurel, D.
Medda, R.
Meyer, L.
Morozova, K. S.
Müller, V.
Myers, J. R.
Noren, C. J.
Olenych, S.
Öz, S.
Patterson, G. H.
Pfeifer, A. C.
Piatkevich, K. D.
Pick, H.
Punge, A.
Ringemann, C.
Rizzoli, S. O.
Rust, M. J.
Schmidt, R.
Schönle, A.
Schulmeister, S.
Schwarz, U.
Sieber, J. J.
Siegberg, D.
Sorkin, A.
Sougrat, R.
Sourjik, V.
Stöhr, K.
Strack, R. L.
Sun, L.
Sun, X.
Szent-Gyorgyi, C.
Toomre, D.
Verkhusha, V. V.
Vicidomini, G.
Vogel, H.
Waggoner, A. S.
Westphal, V.
Wichmann, J.
Wildanger, D.
Willig, K. I.
Woolford, C. A.
Wurm, C. A.
Xu, M.-Q.
Yan, Q.
Zhang, A.
Zhang, J.
Zhuang, X.
- A. Keppler, H. Pick, C. Arrivoli, H. Vogel, and K. Johnsson, “Labeling of fusion proteins with synthetic fluorophores in live cells,” Proc. Natl. Acad. Sci. U.S.A. 101(27), 9955–9959 (2004). [CrossRef] [PubMed]
- K. Stöhr, D. Siegberg, T. Ehrhard, K. Lymperopoulos, S. Öz, S. Schulmeister, A. C. Pfeifer, J. Bachmann, U. Klingmüller, V. Sourjik, and D. P. Herten, “Quenched substrates for live-cell labeling of SNAP-tagged fusion proteins with improved fluorescent background,” Anal. Chem. 82(19), 8186–8193 (2010). [CrossRef] [PubMed]
- X. Sun, A. Zhang, B. Baker, L. Sun, A. Howard, J. Buswell, D. Maurel, A. Masharina, K. Johnsson, C. J. Noren, M.-Q. Xu, and I. R. Corrêa., “Development of SNAP-tag fluorogenic probes for wash-free fluoescence imaging,” ChemBioChem . in press. [PubMed]
- M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
- A. Gautier, A. Juillerat, C. Heinis, I. R. Corrêa, M. Kindermann, F. Beaufils, and K. Johnsson, “An engineered protein tag for multiprotein labeling in living cells,” Chem. Biol. 15(2), 128–136 (2008). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, J. Bierwagen, C. Ringemann, V. Müller, C. Eggeling, and S. W. Hell, “Red-emitting rhodamine dyes for fluorescence microscopy and nanoscopy,” Chemistry 16(1), 158–166 (2010). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, C. A. Wurm, B. Harke, M. Leutenegger, C. Eggeling, and S. W. Hell, “A versatile route to red-emitting carbopyronine dyes for optical microscopy and nanoscopy,” Eur. J. Org. Chem. 2010(19), 3593–3610 (2010). [CrossRef]
- J. A. J. Fitzpatrick, Q. Yan, J. J. Sieber, M. Dyba, U. Schwarz, C. Szent-Gyorgyi, C. A. Woolford, P. B. Berget, A. S. Waggoner, and M. P. Bruchez, “STED nanoscopy in living cells using Fluorogen Activating Proteins,” Bioconjug. Chem. 20(10), 1843–1847 (2009). [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]
- T. J. Gould, J. R. Myers, and J. Bewersdorf, “Total internal reflection STED microscopy,” Opt. Express 19(14), 13351–13357 (2011). [CrossRef] [PubMed]
- D. Toomre and J. Bewersdorf, “A new wave of cellular imaging,” Annu. Rev. Cell Dev. Biol. 26(1), 285–314 (2010). [CrossRef] [PubMed]
- K. S. Morozova, K. D. Piatkevich, T. J. Gould, J. Zhang, J. Bewersdorf, and V. V. Verkhusha, “Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy,” Biophys. J. 99(2), L13–L15 (2010). [CrossRef] [PubMed]
- R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan, and B. S. Glick, “A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling,” Biochemistry 48(35), 8279–8281 (2009). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, J. Bierwagen, C. Ringemann, V. Müller, C. Eggeling, and S. W. Hell, “Red-emitting rhodamine dyes for fluorescence microscopy and nanoscopy,” Chemistry 16(1), 158–166 (2010). [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]
- J. A. J. Fitzpatrick, Q. Yan, J. J. Sieber, M. Dyba, U. Schwarz, C. Szent-Gyorgyi, C. A. Woolford, P. B. Berget, A. S. Waggoner, and M. P. Bruchez, “STED nanoscopy in living cells using Fluorogen Activating Proteins,” Bioconjug. Chem. 20(10), 1843–1847 (2009). [CrossRef] [PubMed]
- X. Sun, A. Zhang, B. Baker, L. Sun, A. Howard, J. Buswell, D. Maurel, A. Masharina, K. Johnsson, C. J. Noren, M.-Q. Xu, and I. R. Corrêa., “Development of SNAP-tag fluorogenic probes for wash-free fluoescence imaging,” ChemBioChem . in press. [PubMed]
- A. Sorkin and G. Carpenter, “Dimerization of internalized epidermal growth factor receptors,” J. Biol. Chem. 266(34), 23453–23460 (1991). [PubMed]
- R. E. Carter and A. Sorkin, “Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera,” J. Biol. Chem. 273(52), 35000–35007 (1998). [CrossRef] [PubMed]
- A. Gautier, A. Juillerat, C. Heinis, I. R. Corrêa, M. Kindermann, F. Beaufils, and K. Johnsson, “An engineered protein tag for multiprotein labeling in living cells,” Chem. Biol. 15(2), 128–136 (2008). [CrossRef] [PubMed]
- X. Sun, A. Zhang, B. Baker, L. Sun, A. Howard, J. Buswell, D. Maurel, A. Masharina, K. Johnsson, C. J. Noren, M.-Q. Xu, and I. R. Corrêa., “Development of SNAP-tag fluorogenic probes for wash-free fluoescence imaging,” ChemBioChem . in press. [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]
- L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, and S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (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. A. J. Fitzpatrick, Q. Yan, J. J. Sieber, M. Dyba, U. Schwarz, C. Szent-Gyorgyi, C. A. Woolford, P. B. Berget, A. S. Waggoner, and M. P. Bruchez, “STED nanoscopy in living cells using Fluorogen Activating Proteins,” Bioconjug. Chem. 20(10), 1843–1847 (2009). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, C. A. Wurm, B. Harke, M. Leutenegger, C. Eggeling, and S. W. Hell, “A versatile route to red-emitting carbopyronine dyes for optical microscopy and nanoscopy,” Eur. J. Org. Chem. 2010(19), 3593–3610 (2010). [CrossRef]
- K. Kolmakov, V. N. Belov, J. Bierwagen, C. Ringemann, V. Müller, C. Eggeling, and S. W. Hell, “Red-emitting rhodamine dyes for fluorescence microscopy and nanoscopy,” Chemistry 16(1), 158–166 (2010). [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]
- R. Schmidt, C. A. Wurm, A. Punge, A. Egner, S. Jakobs, and S. W. Hell, “Mitochondrial cristae revealed with focused light,” Nano Lett. 9(6), 2508–2510 (2009). [CrossRef] [PubMed]
- R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, and S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008). [CrossRef] [PubMed]
- K. Stöhr, D. Siegberg, T. Ehrhard, K. Lymperopoulos, S. Öz, S. Schulmeister, A. C. Pfeifer, J. Bachmann, U. Klingmüller, V. Sourjik, and D. P. Herten, “Quenched substrates for live-cell labeling of SNAP-tagged fusion proteins with improved fluorescent background,” Anal. Chem. 82(19), 8186–8193 (2010). [CrossRef] [PubMed]
- R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, and S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008). [CrossRef] [PubMed]
- J. A. J. Fitzpatrick, Q. Yan, J. J. Sieber, M. Dyba, U. Schwarz, C. Szent-Gyorgyi, C. A. Woolford, P. B. Berget, A. S. Waggoner, and M. P. Bruchez, “STED nanoscopy in living cells using Fluorogen Activating Proteins,” Bioconjug. Chem. 20(10), 1843–1847 (2009). [CrossRef] [PubMed]
- A. Gautier, A. Juillerat, C. Heinis, I. R. Corrêa, M. Kindermann, F. Beaufils, and K. Johnsson, “An engineered protein tag for multiprotein labeling in living cells,” Chem. Biol. 15(2), 128–136 (2008). [CrossRef] [PubMed]
- A. Keppler, S. Gendreizig, T. Gronemeyer, H. Pick, H. Vogel, and K. Johnsson, “A general method for the covalent labeling of fusion proteins with small molecules in vivo,” Nat. Biotechnol. 21(1), 86–89 (2003). [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]
- R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan, and B. S. Glick, “A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling,” Biochemistry 48(35), 8279–8281 (2009). [CrossRef] [PubMed]
- T. J. Gould, J. R. Myers, and J. Bewersdorf, “Total internal reflection STED microscopy,” Opt. Express 19(14), 13351–13357 (2011). [CrossRef] [PubMed]
- K. S. Morozova, K. D. Piatkevich, T. J. Gould, J. Zhang, J. Bewersdorf, and V. V. Verkhusha, “Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy,” Biophys. J. 99(2), L13–L15 (2010). [CrossRef] [PubMed]
- A. Keppler, S. Gendreizig, T. Gronemeyer, H. Pick, H. Vogel, and K. Johnsson, “A general method for the covalent labeling of fusion proteins with small molecules in vivo,” Nat. Biotechnol. 21(1), 86–89 (2003). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, C. A. Wurm, B. Harke, M. Leutenegger, C. Eggeling, and S. W. Hell, “A versatile route to red-emitting carbopyronine dyes for optical microscopy and nanoscopy,” Eur. J. Org. Chem. 2010(19), 3593–3610 (2010). [CrossRef]
- B. Hein, K. I. Willig, C. A. Wurm, V. Westphal, S. Jakobs, and S. W. Hell, “Stimulated emission depletion nanoscopy of living cells using SNAP-tag fusion proteins,” Biophys. J. 98(1), 158–163 (2010). [CrossRef] [PubMed]
- R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan, and B. S. Glick, “A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling,” Biochemistry 48(35), 8279–8281 (2009). [CrossRef] [PubMed]
- A. Gautier, A. Juillerat, C. Heinis, I. R. Corrêa, M. Kindermann, F. Beaufils, and K. Johnsson, “An engineered protein tag for multiprotein labeling in living cells,” Chem. Biol. 15(2), 128–136 (2008). [CrossRef] [PubMed]
- J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, and S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, C. A. Wurm, B. Harke, M. Leutenegger, C. Eggeling, and S. W. Hell, “A versatile route to red-emitting carbopyronine dyes for optical microscopy and nanoscopy,” Eur. J. Org. Chem. 2010(19), 3593–3610 (2010). [CrossRef]
- K. Kolmakov, V. N. Belov, J. Bierwagen, C. Ringemann, V. Müller, C. Eggeling, and S. W. Hell, “Red-emitting rhodamine dyes for fluorescence microscopy and nanoscopy,” Chemistry 16(1), 158–166 (2010). [CrossRef] [PubMed]
- B. Hein, K. I. Willig, C. A. Wurm, V. Westphal, S. Jakobs, and S. W. Hell, “Stimulated emission depletion nanoscopy of living cells using SNAP-tag fusion proteins,” Biophys. J. 98(1), 158–163 (2010). [CrossRef] [PubMed]
- S. W. Hell, “Microscopy and its focal switch,” Nat. Methods 6(1), 24–32 (2009). [CrossRef] [PubMed]
- R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan, and B. S. Glick, “A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling,” Biochemistry 48(35), 8279–8281 (2009). [CrossRef] [PubMed]
- R. Schmidt, C. A. Wurm, A. Punge, A. Egner, S. Jakobs, and S. W. Hell, “Mitochondrial cristae revealed with focused light,” Nano Lett. 9(6), 2508–2510 (2009). [CrossRef] [PubMed]
- L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, and S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008). [CrossRef] [PubMed]
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- K. S. Morozova, K. D. Piatkevich, T. J. Gould, J. Zhang, J. Bewersdorf, and V. V. Verkhusha, “Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy,” Biophys. J. 99(2), L13–L15 (2010). [CrossRef] [PubMed]
- M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
Anal. Chem.
- K. Stöhr, D. Siegberg, T. Ehrhard, K. Lymperopoulos, S. Öz, S. Schulmeister, A. C. Pfeifer, J. Bachmann, U. Klingmüller, V. Sourjik, and D. P. Herten, “Quenched substrates for live-cell labeling of SNAP-tagged fusion proteins with improved fluorescent background,” Anal. Chem. 82(19), 8186–8193 (2010). [CrossRef] [PubMed]
Annu. Rev. Cell Dev. Biol.
- D. Toomre and J. Bewersdorf, “A new wave of cellular imaging,” Annu. Rev. Cell Dev. Biol. 26(1), 285–314 (2010). [CrossRef] [PubMed]
Biochemistry
- R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan, and B. S. Glick, “A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling,” Biochemistry 48(35), 8279–8281 (2009). [CrossRef] [PubMed]
Bioconjug. Chem.
- J. A. J. Fitzpatrick, Q. Yan, J. J. Sieber, M. Dyba, U. Schwarz, C. Szent-Gyorgyi, C. A. Woolford, P. B. Berget, A. S. Waggoner, and M. P. Bruchez, “STED nanoscopy in living cells using Fluorogen Activating Proteins,” Bioconjug. Chem. 20(10), 1843–1847 (2009). [CrossRef] [PubMed]
Biophys. J.
- B. Hein, K. I. Willig, C. A. Wurm, V. Westphal, S. Jakobs, and S. W. Hell, “Stimulated emission depletion nanoscopy of living cells using SNAP-tag fusion proteins,” Biophys. J. 98(1), 158–163 (2010). [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]
- 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. S. Morozova, K. D. Piatkevich, T. J. Gould, J. Zhang, J. Bewersdorf, and V. V. Verkhusha, “Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy,” Biophys. J. 99(2), L13–L15 (2010). [CrossRef] [PubMed]
Chem. Biol.
- A. Gautier, A. Juillerat, C. Heinis, I. R. Corrêa, M. Kindermann, F. Beaufils, and K. Johnsson, “An engineered protein tag for multiprotein labeling in living cells,” Chem. Biol. 15(2), 128–136 (2008). [CrossRef] [PubMed]
ChemBioChem
- X. Sun, A. Zhang, B. Baker, L. Sun, A. Howard, J. Buswell, D. Maurel, A. Masharina, K. Johnsson, C. J. Noren, M.-Q. Xu, and I. R. Corrêa., “Development of SNAP-tag fluorogenic probes for wash-free fluoescence imaging,” ChemBioChem . in press. [PubMed]
Chemistry
- K. Kolmakov, V. N. Belov, J. Bierwagen, C. Ringemann, V. Müller, C. Eggeling, and S. W. Hell, “Red-emitting rhodamine dyes for fluorescence microscopy and nanoscopy,” Chemistry 16(1), 158–166 (2010). [CrossRef] [PubMed]
Eur. J. Org. Chem.
- K. Kolmakov, V. N. Belov, C. A. Wurm, B. Harke, M. Leutenegger, C. Eggeling, and S. W. Hell, “A versatile route to red-emitting carbopyronine dyes for optical microscopy and nanoscopy,” Eur. J. Org. Chem. 2010(19), 3593–3610 (2010). [CrossRef]
J. Biol. Chem.
- R. E. Carter and A. Sorkin, “Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera,” J. Biol. Chem. 273(52), 35000–35007 (1998). [CrossRef] [PubMed]
- A. Sorkin and G. Carpenter, “Dimerization of internalized epidermal growth factor receptors,” J. Biol. Chem. 266(34), 23453–23460 (1991). [PubMed]
Nano Lett.
- R. Schmidt, C. A. Wurm, A. Punge, A. Egner, S. Jakobs, and S. W. Hell, “Mitochondrial cristae revealed with focused light,” Nano Lett. 9(6), 2508–2510 (2009). [CrossRef] [PubMed]
Nat. Biotechnol.
- A. Keppler, S. Gendreizig, T. Gronemeyer, H. Pick, H. Vogel, and K. Johnsson, “A general method for the covalent labeling of fusion proteins with small molecules in vivo,” Nat. Biotechnol. 21(1), 86–89 (2003). [CrossRef] [PubMed]
Nat. Methods
- M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
- S. W. Hell, “Microscopy and its focal switch,” Nat. Methods 6(1), 24–32 (2009). [CrossRef] [PubMed]
- R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, and S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008). [CrossRef] [PubMed]
Opt. Express
- T. J. Gould, J. R. Myers, and J. Bewersdorf, “Total internal reflection STED microscopy,” Opt. Express 19(14), 13351–13357 (2011). [CrossRef] [PubMed]
- J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, and S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011). [CrossRef] [PubMed]
Opt. Lett.
- 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]
Proc. Natl. Acad. Sci. U.S.A.
- A. Keppler, H. Pick, C. Arrivoli, H. Vogel, and K. Johnsson, “Labeling of fusion proteins with synthetic fluorophores in live cells,” Proc. Natl. Acad. Sci. U.S.A. 101(27), 9955–9959 (2004). [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]
Small
- L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, and S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008). [CrossRef] [PubMed]
2011, Bückers, Opt. Express
- D. Toomre and J. Bewersdorf, “A new wave of cellular imaging,” Annu. Rev. Cell Dev. Biol. 26(1), 285–314 (2010). [CrossRef] [PubMed]
- B. Hein, K. I. Willig, C. A. Wurm, V. Westphal, S. Jakobs, and S. W. Hell, “Stimulated emission depletion nanoscopy of living cells using SNAP-tag fusion proteins,” Biophys. J. 98(1), 158–163 (2010). [CrossRef] [PubMed]
- K. Stöhr, D. Siegberg, T. Ehrhard, K. Lymperopoulos, S. Öz, S. Schulmeister, A. C. Pfeifer, J. Bachmann, U. Klingmüller, V. Sourjik, and D. P. Herten, “Quenched substrates for live-cell labeling of SNAP-tagged fusion proteins with improved fluorescent background,” Anal. Chem. 82(19), 8186–8193 (2010). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, J. Bierwagen, C. Ringemann, V. Müller, C. Eggeling, and S. W. Hell, “Red-emitting rhodamine dyes for fluorescence microscopy and nanoscopy,” Chemistry 16(1), 158–166 (2010). [CrossRef] [PubMed]
- K. Kolmakov, V. N. Belov, C. A. Wurm, B. Harke, M. Leutenegger, C. Eggeling, and S. W. Hell, “A versatile route to red-emitting carbopyronine dyes for optical microscopy and nanoscopy,” Eur. J. Org. Chem. 2010(19), 3593–3610 (2010). [CrossRef]
- K. S. Morozova, K. D. Piatkevich, T. J. Gould, J. Zhang, J. Bewersdorf, and V. V. Verkhusha, “Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy,” Biophys. J. 99(2), L13–L15 (2010). [CrossRef] [PubMed]
- R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan, and B. S. Glick, “A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling,” Biochemistry 48(35), 8279–8281 (2009). [CrossRef] [PubMed]
- S. W. Hell, “Microscopy and its focal switch,” Nat. Methods 6(1), 24–32 (2009). [CrossRef] [PubMed]
- J. A. J. Fitzpatrick, Q. Yan, J. J. Sieber, M. Dyba, U. Schwarz, C. Szent-Gyorgyi, C. A. Woolford, P. B. Berget, A. S. Waggoner, and M. P. Bruchez, “STED nanoscopy in living cells using Fluorogen Activating Proteins,” Bioconjug. Chem. 20(10), 1843–1847 (2009). [CrossRef] [PubMed]
- R. Schmidt, C. A. Wurm, A. Punge, A. Egner, S. Jakobs, and S. W. Hell, “Mitochondrial cristae revealed with focused light,” Nano Lett. 9(6), 2508–2510 (2009). [CrossRef] [PubMed]
- A. Gautier, A. Juillerat, C. Heinis, I. R. Corrêa, M. Kindermann, F. Beaufils, and K. Johnsson, “An engineered protein tag for multiprotein labeling in living cells,” Chem. Biol. 15(2), 128–136 (2008). [CrossRef] [PubMed]
- L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, and S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008). [CrossRef] [PubMed]
- R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, and S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008). [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]
- 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]
- 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. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
- A. Keppler, H. Pick, C. Arrivoli, H. Vogel, and K. Johnsson, “Labeling of fusion proteins with synthetic fluorophores in live cells,” Proc. Natl. Acad. Sci. U.S.A. 101(27), 9955–9959 (2004). [CrossRef] [PubMed]
- A. Keppler, S. Gendreizig, T. Gronemeyer, H. Pick, H. Vogel, and K. Johnsson, “A general method for the covalent labeling of fusion proteins with small molecules in vivo,” Nat. Biotechnol. 21(1), 86–89 (2003). [CrossRef] [PubMed]
- R. E. Carter and A. Sorkin, “Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera,” J. Biol. Chem. 273(52), 35000–35007 (1998). [CrossRef] [PubMed]
- A. Sorkin and G. Carpenter, “Dimerization of internalized epidermal growth factor receptors,” J. Biol. Chem. 266(34), 23453–23460 (1991). [PubMed]
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