Abstract

We describe the design and implementation of a stimulated emission depletion (STED) microscope which allows simultaneous three-dimensional super-resolution imaging in two colors. A super-continuum laser source is used to provide all spectral bands necessary for excitation and efficient depletion to achieve a lateral and axial resolution of ~35 nm and ~90 nm respectively. We characterize the systems' performance by imaging colloidal particles and single fluorescent molecules. Its biological applicability is demonstrated by dual-color imaging of nuclear pore complexes and of DNA replication sites in mammalian cells.

© 2014 Optical Society of America

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2013 (2)

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

2012 (3)

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

C. Coltharp, J. Xiao, “Superresolution microscopy for microbiology,” Cell. Microbiol. 14(12), 1808–1818 (2012).
[CrossRef] [PubMed]

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

2011 (3)

2010 (4)

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

L. Schermelleh, R. Heintzmann, H. Leonhardt, “A guide to super-resolution fluorescence microscopy,” J. Cell Biol. 190(2), 165–175 (2010).
[CrossRef] [PubMed]

D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 4 (2010).
[CrossRef] [PubMed]

2009 (2)

D. Wildanger, R. Medda, L. Kastrup, S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009).
[CrossRef] [PubMed]

Z. Cseresnyes, U. Schwarz, C. M. Green, “Analysis of replication factories in human cells by super-resolution light microscopy,” BMC Cell Biol. 10(1), 88 (2009).
[CrossRef] [PubMed]

2008 (5)

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

D. Wildanger, E. Rittweger, L. Kastrup, S. W. Hell, “STED microscopy with a supercontinuum laser source,” Opt. Express 16(13), 9614–9621 (2008).
[CrossRef] [PubMed]

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, 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, S. W. Hell, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement,” Science 320(5873), 246–249 (2008).
[CrossRef] [PubMed]

1998 (1)

M. Nagorni, S. W. Hell, “4Pi-confocal microscopy provides three-dimensional images of the microtubule network with 100- to 150-nm resolution,” J. Struct. Biol. 123(3), 236–247 (1998).
[CrossRef] [PubMed]

1994 (1)

Agard, D. A.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Arganda-Carreras, I.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Baddeley, D.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Berning, S.

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

Birk, U.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Bönisch, C.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Briggs, J. A.

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

Bückers, J.

J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011).
[CrossRef] [PubMed]

D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 4 (2010).
[CrossRef] [PubMed]

Burke, B.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Cardona, A.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Cardoso, M. C.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Carlton, P. M.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Chagin, V. O.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Coltharp, C.

C. Coltharp, J. Xiao, “Superresolution microscopy for microbiology,” Cell. Microbiol. 14(12), 1808–1818 (2012).
[CrossRef] [PubMed]

Cordes, V. C.

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

Cremer, C.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Cseresnyes, Z.

Z. Cseresnyes, U. Schwarz, C. M. Green, “Analysis of replication factories in human cells by super-resolution light microscopy,” BMC Cell Biol. 10(1), 88 (2009).
[CrossRef] [PubMed]

Dabauvalle, M. C.

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

Daigle, N.

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

de Marco, A.

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

Domaing, P.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Donnert, G.

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

Eggeling, C.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

Egner, A.

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008).
[CrossRef] [PubMed]

Eliceiri, K.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Ellenberg, J.

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

Engelhardt, J.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008).
[CrossRef] [PubMed]

Frise, E.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Gahl, A.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Göttfert, F.

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

Green, C. M.

Z. Cseresnyes, U. Schwarz, C. M. Green, “Analysis of replication factories in human cells by super-resolution light microscopy,” BMC Cell Biol. 10(1), 88 (2009).
[CrossRef] [PubMed]

Gustafsson, M. G.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Haase, S.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Hake, S. B.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Han, K. Y.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

Hartenstein, V.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Heilemann, M.

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

Heintzmann, R.

L. Schermelleh, R. Heintzmann, H. Leonhardt, “A guide to super-resolution fluorescence microscopy,” J. Cell Biol. 190(2), 165–175 (2010).
[CrossRef] [PubMed]

Hell, S. W.

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011).
[CrossRef] [PubMed]

D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 4 (2010).
[CrossRef] [PubMed]

D. Wildanger, R. Medda, L. Kastrup, S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009).
[CrossRef] [PubMed]

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008).
[CrossRef] [PubMed]

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, 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, S. W. Hell, “STED microscopy with a supercontinuum laser source,” Opt. Express 16(13), 9614–9621 (2008).
[CrossRef] [PubMed]

M. Nagorni, S. W. Hell, “4Pi-confocal microscopy provides three-dimensional images of the microtubule network with 100- to 150-nm resolution,” J. Struct. Biol. 123(3), 236–247 (1998).
[CrossRef] [PubMed]

S. W. Hell, J. Wichmann, “Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy,” Opt. Lett. 19(11), 780–782 (1994).
[CrossRef] [PubMed]

Honigmann, A.

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

Israel, L.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Jahn, R.

V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, S. W. Hell, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement,” Science 320(5873), 246–249 (2008).
[CrossRef] [PubMed]

Jakobs, S.

D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 4 (2010).
[CrossRef] [PubMed]

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008).
[CrossRef] [PubMed]

Kamin, D.

V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, S. W. Hell, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement,” Science 320(5873), 246–249 (2008).
[CrossRef] [PubMed]

Kastrup, L.

J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011).
[CrossRef] [PubMed]

D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 4 (2010).
[CrossRef] [PubMed]

D. Wildanger, R. Medda, L. Kastrup, 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, S. W. Hell, “STED microscopy with a supercontinuum laser source,” Opt. Express 16(13), 9614–9621 (2008).
[CrossRef] [PubMed]

Kaynig, V.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Kner, P.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Kremmer, E.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Krohne, G.

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

Lauterbach, M. A.

V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, S. W. Hell, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement,” Science 320(5873), 246–249 (2008).
[CrossRef] [PubMed]

Leonhardt, H.

L. Schermelleh, R. Heintzmann, H. Leonhardt, “A guide to super-resolution fluorescence microscopy,” J. Cell Biol. 190(2), 165–175 (2010).
[CrossRef] [PubMed]

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Longair, M.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Löschberger, A.

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

Maiser, A.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Martin, S.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Matheisl, S.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Medda, R.

D. Wildanger, R. Medda, L. Kastrup, S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009).
[CrossRef] [PubMed]

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

Merkl, R.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Meyer, L.

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

Michaelis, J.

Mildner, S. N.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Moffitt, J. R.

Moneron, G.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

Mueller, V.

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

Nagorni, M.

M. Nagorni, S. W. Hell, “4Pi-confocal microscopy provides three-dimensional images of the microtubule network with 100- to 150-nm resolution,” J. Struct. Biol. 123(3), 236–247 (1998).
[CrossRef] [PubMed]

Neumann, D.

D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 4 (2010).
[CrossRef] [PubMed]

Osseforth, C.

Pietzsch, T.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Pombo, A.

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Preibisch, S.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Punge, A.

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

Reuss, M.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

Rieger, B.

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

Rittweger, E.

Rizzoli, S. O.

V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, S. W. Hell, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement,” Science 320(5873), 246–249 (2008).
[CrossRef] [PubMed]

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

Rueden, C.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Saalfeld, S.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Sauer, M.

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

Schermelleh, L.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

L. Schermelleh, R. Heintzmann, H. Leonhardt, “A guide to super-resolution fluorescence microscopy,” J. Cell Biol. 190(2), 165–175 (2010).
[CrossRef] [PubMed]

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Schindelin, J.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Schmid, B.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Schmidt, R.

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008).
[CrossRef] [PubMed]

Schwarz, U.

Z. Cseresnyes, U. Schwarz, C. M. Green, “Analysis of replication factories in human cells by super-resolution light microscopy,” BMC Cell Biol. 10(1), 88 (2009).
[CrossRef] [PubMed]

Sedat, J. W.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Shao, L.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Straub, T.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Szymborska, A.

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

Ta, H.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

Tinevez, J.-Y.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Tomancak, P.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

van de Linde, S.

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

Vicidomini, G.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011).
[CrossRef] [PubMed]

Westphal, V.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, S. W. Hell, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement,” Science 320(5873), 246–249 (2008).
[CrossRef] [PubMed]

White, D. J.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

Wichmann, J.

Wiedemann, S. M.

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

Wildanger, D.

J. Bückers, D. Wildanger, G. Vicidomini, L. Kastrup, S. W. Hell, “Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses,” Opt. Express 19(4), 3130–3143 (2011).
[CrossRef] [PubMed]

D. Wildanger, R. Medda, L. Kastrup, S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009).
[CrossRef] [PubMed]

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

D. Wildanger, E. Rittweger, L. Kastrup, S. W. Hell, “STED microscopy with a supercontinuum laser source,” Opt. Express 16(13), 9614–9621 (2008).
[CrossRef] [PubMed]

Winoto, L.

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

Wurm, C. A.

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008).
[CrossRef] [PubMed]

Xiao, J.

C. Coltharp, J. Xiao, “Superresolution microscopy for microbiology,” Cell. Microbiol. 14(12), 1808–1818 (2012).
[CrossRef] [PubMed]

Biophys. J. (1)

F. Göttfert, C. A. Wurm, V. Mueller, S. Berning, V. C. Cordes, A. Honigmann, S. W. Hell, “Coaligned Dual-Channel STED Nanoscopy and Molecular Diffusion Analysis at 20 nm Resolution,” Biophys. J. 105(1), L01–L03 (2013).
[CrossRef] [PubMed]

BMC Cell Biol. (1)

Z. Cseresnyes, U. Schwarz, C. M. Green, “Analysis of replication factories in human cells by super-resolution light microscopy,” BMC Cell Biol. 10(1), 88 (2009).
[CrossRef] [PubMed]

Cell. Microbiol. (1)

C. Coltharp, J. Xiao, “Superresolution microscopy for microbiology,” Cell. Microbiol. 14(12), 1808–1818 (2012).
[CrossRef] [PubMed]

J. Cell Biol. (2)

L. Schermelleh, R. Heintzmann, H. Leonhardt, “A guide to super-resolution fluorescence microscopy,” J. Cell Biol. 190(2), 165–175 (2010).
[CrossRef] [PubMed]

S. M. Wiedemann, S. N. Mildner, C. Bönisch, L. Israel, A. Maiser, S. Matheisl, T. Straub, R. Merkl, H. Leonhardt, E. Kremmer, L. Schermelleh, S. B. Hake, “Identification and characterization of two novel primate-specific histone H3 variants, H3.X and H3.Y,” J. Cell Biol. 190(5), 777–791 (2010).
[CrossRef] [PubMed]

J. Cell Sci. (1)

A. Löschberger, S. van de Linde, M. C. Dabauvalle, B. Rieger, M. Heilemann, G. Krohne, M. Sauer, “Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution,” J. Cell Sci. 125(3), 570–575 (2012).
[CrossRef] [PubMed]

J. Microsc. (1)

D. Wildanger, R. Medda, L. Kastrup, S. W. Hell, “A compact STED microscope providing 3D nanoscale resolution,” J. Microsc. 236(1), 35–43 (2009).
[CrossRef] [PubMed]

J. Struct. Biol. (1)

M. Nagorni, S. W. Hell, “4Pi-confocal microscopy provides three-dimensional images of the microtubule network with 100- to 150-nm resolution,” J. Struct. Biol. 123(3), 236–247 (1998).
[CrossRef] [PubMed]

Nat. Methods (3)

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods 8(7), 571–573 (2011).
[CrossRef] [PubMed]

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[CrossRef] [PubMed]

R. Schmidt, C. A. Wurm, S. Jakobs, J. Engelhardt, A. Egner, S. W. Hell, “Spherical nanosized focal spot unravels the interior of cells,” Nat. Methods 5(6), 539–544 (2008).
[CrossRef] [PubMed]

Nucleic Acids Res. (1)

D. Baddeley, V. O. Chagin, L. Schermelleh, S. Martin, A. Pombo, P. M. Carlton, A. Gahl, P. Domaing, U. Birk, H. Leonhardt, C. Cremer, M. C. Cardoso, “Measurement of replication structures at the nanometer scale using super-resolution light microscopy,” Nucleic Acids Res. 38(2), e8 (2010).
[CrossRef] [PubMed]

Opt. Express (3)

Opt. Lett. (1)

PMC Biophys (1)

D. Neumann, J. Bückers, L. Kastrup, S. W. Hell, S. Jakobs, “Two-color STED microscopy reveals different degrees of colocalization between hexokinase-I and the three human VDAC isoforms,” PMC Biophys 3(1), 4 (2010).
[CrossRef] [PubMed]

Science (3)

L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. Gustafsson, H. Leonhardt, J. W. Sedat, “Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy,” Science 320(5881), 1332–1336 (2008).
[CrossRef] [PubMed]

V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, S. W. Hell, “Video-rate far-field optical nanoscopy dissects synaptic vesicle movement,” Science 320(5873), 246–249 (2008).
[CrossRef] [PubMed]

A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. Briggs, J. Ellenberg, “Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging,” Science 341(6146), 655–658 (2013).
[CrossRef] [PubMed]

Small (1)

L. Meyer, D. Wildanger, R. Medda, A. Punge, S. O. Rizzoli, G. Donnert, S. W. Hell, “Dual-color STED microscopy at 30-nm focal-plane resolution,” Small 4(8), 1095–1100 (2008).
[CrossRef] [PubMed]

Other (1)

Python(x,y) 2.7.5.1,” http://code.google.com/p/pythonxy/ .

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Figures (5)

Fig. 1
Fig. 1

Schematic of experimental setup. The left part shows the laser source and the optical components necessary to prepare a total of six different beams of desired spectral bands for excitation and depletion. The six beams are coupled into six polarization maintaining fibers. The part on the right depicts the actual microscope starting with the output of the six fibers, the components for spatial co-alignment of the various beams, the phase plates for the purpose of shaping the PSF of the depletion beam, the objective lens and the detection beam path for the two fluorescence channels.

Fig. 2
Fig. 2

Experimentally measured point-spread functions (upper panels; green for excitation and magenta for depletion patterns; shown representatively for channel 2) and corresponding fluorescence images (lower panels). Point-spread functions were measured by recording the scattering from an immobilized 80 nm gold bead scanned through the focus. For simplicity, only data from x-z scans are presented showing both the lateral as well as the axial PSF. The fluorescence images were recorded using 100 nm fluorescent beads dispersed on a cover slip (fluorescence images are shown representatively for channel 1). A) and E) Confocal mode using only the excitation beam and resulting in a diffraction limited PSF and fluorescence image which is considerably spread along the z-axis. B) and F) The XY-STED beam improves only lateral resolution; axial resolution is not changed. C) and G) The Z-STED beam acts mainly in the axial dimension. Additionally, resolution is also slightly improved along the lateral dimension. D) and H) By spatially co-aligning the XY and Z STED beams resolution is dramatically enhanced in all three dimensions.

Fig. 3
Fig. 3

Dual-color imaging of a mixture of immobilized DNA molecules labeled with single Atto590 or Atto647N fluorophores. A) Single molecules imaged in STED-2D mode. Green LUT shows fluorescence intensity of Atto590, magenta shows that of Atto647N molecules. Arrows illustrate the position of the cross-sections shown in B) (Atto590) and C) (Atto647N). D) Cross-section along the z-axis of a single Atto647N molecule shows a FWHM of ~83 nm. Image A) was post processed for printing purposes but not for data analysis, fits in B), C) and D) show raw data. Pixel size is 10 nm2 (panels B and C) or 20 nm2 (panel D) at a pixel dwell time of 400 µs. Fitting and plotting was performed using the Python distribution Python(x,y) [12].

Fig. 4
Fig. 4

Dual-color STED microscopy in 2D mode of fixed U2OS cells using antibodies targeting different parts of NPCs. Green: Primary antibody against FG-repeat containing nucleoporins detected via secondary antibody conjugated to Alexa594. Magenta: Primary antibody against inner nuclear pore basket protein Tpr detected via secondary antibody conjugated to Atto647N. Images were background corrected and contrast was enhanced for printing purposes. Pixel dwell times were 180 µs. Depending on the z-height of the imaging plane different views of the cell nucleus are obtained. A) Apical slice shows a co-localization of the two secondary antibodies with a FWHM of single nuclear pore complexes of ~80 nm. B) Mid-section slice through the nucleus shows a clear separation between the two secondary antibodies. In contrast to Tpr, the FG-repeat antibody typically displays two spots, in the central part and the cytoplasmic side of NPCs (white arrows), demonstrating the ability to separate different epitopes within this large ~125 MDa sized complex. C) 3D isosurface reconstruction of 34 slices (19 × 19 µm) of one nucleus atop a maximum intensity projection of the same data using Avizo 8.0.1 (FEI, USA) and Fiji [17]. Inset shows zoomed in volume rendering of individual NPCs.

Fig. 5
Fig. 5

3D reconstruction of replication foci in a C2C12 cell nucleus during late S-phase labeled during a 15 min EdU pulse. EdU incorporation was visualized by covalent coupling of Alexa594 dye via click chemistry (first channel; green). The second channel (magenta) shows immunostaining of PCNA using Atto647N-conjugated secondary antibodies. A) 3D isosurface reconstruction of 38 dual-color image planes taken in 3D mode with a voxel size of 50 nm3 at a pixel dwell time of 400 µs using Avizo 8.0.1 and Fiji [17]. The bottom plane (20 × 20 µm) shows the maximum intensity projection of the data set. B) X-y slice of a different nucleus in early S-phase imaged in 3D mode with a pixel size of 20 nm2 and pixel dwell time of 200 µs. C) Upper panel: Representative x-z-slice assembled from the first channel (EdU) of the image stack used for subfigure A. Lower panel: Gaussian function fitted to raw data of upper panel, position indicated by arrow. D) Upper panel: Representative x-z-slice constructed from the second channel (αPCNA) of the image stack used for subfigure A. Lower part: Gaussian function fitted to raw data of upper panel at position indicated by arrow. Image data was post processed to remove background and improve contrast for printing purposes. Figure C and D uses a multicolor look-up table to better visualize the dynamic range of the images. Fitting and plotting was performed with raw data using the Python distribution Python(x,y) [12].

Tables (2)

Tables Icon

Table 1 Fiber couplers, fibers and collimators used in the 3D dual-channel STED microscope. All parts from Schäfter & Kirchhoff GmbH, Germany unless stated otherwise

Tables Icon

Table 2 Key components used for the custom-built 3D STED microscope

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