Abstract

Laser scanning microscopy allows for three-dimensional imaging of cells with molecular specific labeling. However the spatial resolution of optical microscopy is fundamentally limited by the diffraction of light. In the last two decades many techniques have been introduced to enhance the resolution of laser scanning microscopes. However most of these techniques impose strong constraints on the specimen or rely on complex optical systems. These constraints limit the applicability of resolution improvement to various imaging modalities and sample types. To overcome these limitations, we introduce here a novel approach, which we called Switching LAser Mode (SLAM) microscopy, to enhance resolution and contrast in laser scanning microscopy. SLAM microscopy relies on subtracting images obtained with dark and bright modes, and exploits the smaller dimensions of the dark spot of the azimuthally polarized TE01 mode. With this approach, resolution is improved by a factor of two in confocal microscopy. The technique is not based on complex nonlinear processes and thus requires laser power similar to that used in conventional imaging, minimizing photo-damage. The flexibility of the approach enables retrofitting in commercial confocal and two-photon microscopes and opens avenues for resolution enhancement in fluorescence-independent microscopy.

© 2013 OSA

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2012 (2)

R. Fiolka, L. Shao, E. H. Rego, M. W. Davidson, and M. G. Gustafsson, “Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination,” Proc. Natl. Acad. Sci. U.S.A.109(14), 5311–5315 (2012).
[CrossRef] [PubMed]

S. N. Khonina and I. Golub, “How low can STED go? Comparison of different write-erase beam combinations for stimulated emission depletion microscopy,” J. Opt. Soc. Am. A29(10), 2242–2246 (2012).
[CrossRef] [PubMed]

2011 (4)

Y. Kozawa, T. Hibi, A. Sato, H. Horanai, M. Kurihara, N. Hashimoto, H. Yokoyama, T. Nemoto, and S. Sato, “Lateral resolution enhancement of laser scanning microscopy by a higher-order radially polarized mode beam,” Opt. Express19(17), 15947–15954 (2011).
[CrossRef] [PubMed]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

S. A. Jones, S. H. Shim, J. He, and X. Zhuang, “Fast, three-dimensional super-resolution imaging of live cells,” Nat. Methods8(6), 499–505 (2011).
[CrossRef] [PubMed]

L. Shao, P. Kner, E. H. Rego, and M. G. Gustafsson, “Super-resolution 3D microscopy of live whole cells using structured illumination,” Nat. Methods8(12), 1044–1046 (2011).
[CrossRef] [PubMed]

2010 (2)

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

B. R. Boruah, “Lateral resolution enhancement in confocal microscopy by vectorial aperture engineering,” Appl. Opt.49(4), 701–707 (2010).
[CrossRef] [PubMed]

2009 (6)

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

C. Labrakakis, L. E. Lorenzo, C. Bories, A. Ribeiro-da-Silva, and Y. De Koninck, “Inhibitory coupling between inhibitory interneurons in the spinal cord dorsal horn,” Mol. Pain5(1), 24 (2009).
[CrossRef] [PubMed]

E. Bélanger, S. Bégin, S. Laffray, Y. De Koninck, R. Vallée, and D. Côté, “Quantitative myelin imaging with coherent anti-Stokes Raman scattering microscopy: alleviating the excitation polarization dependence with circularly polarized laser beams,” Opt. Express17(21), 18419–18432 (2009).
[CrossRef] [PubMed]

H. Dehez, M. Piché, and Y. De Koninck, “Enhanced resolution in two-photon imaging using a TM01 laser beam at a dielectric interface,” Opt. Lett.34(23), 3601–3603 (2009).
[CrossRef] [PubMed]

J. Kim, D. C. Kim, and S. H. Back, “Demonstration of high lateral resolution in laser confocal microscopy using annular and radially polarized light,” Microsc. Res. Tech.72(6), 441–446 (2009).
[CrossRef] [PubMed]

O. Haeberlé and B. Simon, “Saturated structured confocal microscopy with theoretically unlimited resolution,” Opt. Commun.282(18), 3657–3664 (2009).
[CrossRef]

2008 (2)

H. Shroff, C. G. Galbraith, J. A. Galbraith, and E. Betzig, “Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics,” Nat. Methods5(5), 417–423 (2008).
[CrossRef] [PubMed]

J. Stadler, C. Stanciu, C. Stupperich, and A. J. Meixner, “Tighter focusing with a parabolic mirror,” Opt. Lett.33(7), 681–683 (2008).
[CrossRef] [PubMed]

2006 (5)

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,” Science313(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. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

S. T. Hess, T. P. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

2005 (2)

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

H. Balci, T. Ha, H. L. Sweeney, and P. R. Selvin, “Interhead distance measurements in myosin VI via SHRImP support a simplified hand-over-hand model,” Biophys. J.89(1), 413–417 (2005).
[CrossRef] [PubMed]

2003 (3)

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[CrossRef] [PubMed]

T. A. Pologruto, B. L. Sabatini, and K. Svoboda, “ScanImage: flexible software for operating laser scanning microscopes,” Biomed. Eng. Online2(1), 13 (2003).
[CrossRef] [PubMed]

R. Dorn, S. Quabis, and G. Leuchs, “Sharper focus for a radially polarized light beam,” Phys. Rev. Lett.91(23), 233901 (2003).
[CrossRef] [PubMed]

Andrei, M. A.

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Back, S. H.

J. Kim, D. C. Kim, and S. H. Back, “Demonstration of high lateral resolution in laser confocal microscopy using annular and radially polarized light,” Microsc. Res. Tech.72(6), 441–446 (2009).
[CrossRef] [PubMed]

Balci, H.

H. Balci, T. Ha, H. L. Sweeney, and P. R. Selvin, “Interhead distance measurements in myosin VI via SHRImP support a simplified hand-over-hand model,” Biophys. J.89(1), 413–417 (2005).
[CrossRef] [PubMed]

Bates, M.

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

Beaurepaire, E.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Bégin, S.

Bélanger, E.

Betzig, E.

H. Shroff, C. G. Galbraith, J. A. Galbraith, and E. Betzig, “Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics,” Nat. Methods5(5), 417–423 (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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Bonifacino, J. S.

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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Bories, C.

C. Labrakakis, L. E. Lorenzo, C. Bories, A. Ribeiro-da-Silva, and Y. De Koninck, “Inhibitory coupling between inhibitory interneurons in the spinal cord dorsal horn,” Mol. Pain5(1), 24 (2009).
[CrossRef] [PubMed]

Boruah, B. R.

Combettes, L.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Côté, D.

Davidson, M. W.

R. Fiolka, L. Shao, E. H. Rego, M. W. Davidson, and M. G. Gustafsson, “Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination,” Proc. Natl. Acad. Sci. U.S.A.109(14), 5311–5315 (2012).
[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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

De Koninck, P.

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

De Koninck, Y.

Débarre, D.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Dehez, H.

Deuschle, K.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Donnert, G.

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Dorn, R.

R. Dorn, S. Quabis, and G. Leuchs, “Sharper focus for a radially polarized light beam,” Phys. Rev. Lett.91(23), 233901 (2003).
[CrossRef] [PubMed]

Eggeling, C.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Engelhardt, J.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

Fabre, A.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Fiolka, R.

R. Fiolka, L. Shao, E. H. Rego, M. W. Davidson, and M. G. Gustafsson, “Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination,” Proc. Natl. Acad. Sci. U.S.A.109(14), 5311–5315 (2012).
[CrossRef] [PubMed]

Forkey, J. N.

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[CrossRef] [PubMed]

Forthmann, C.

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

Galbraith, C. G.

H. Shroff, C. G. Galbraith, J. A. Galbraith, and E. Betzig, “Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics,” Nat. Methods5(5), 417–423 (2008).
[CrossRef] [PubMed]

Galbraith, J. A.

H. Shroff, C. G. Galbraith, J. A. Galbraith, and E. Betzig, “Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics,” Nat. Methods5(5), 417–423 (2008).
[CrossRef] [PubMed]

Girirajan, T. P.

S. T. Hess, T. P. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

Goldman, Y. E.

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[CrossRef] [PubMed]

Golub, I.

Gustafsson, M. G.

R. Fiolka, L. Shao, E. H. Rego, M. W. Davidson, and M. G. Gustafsson, “Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination,” Proc. Natl. Acad. Sci. U.S.A.109(14), 5311–5315 (2012).
[CrossRef] [PubMed]

L. Shao, P. Kner, E. H. Rego, and M. G. Gustafsson, “Super-resolution 3D microscopy of live whole cells using structured illumination,” Nat. Methods8(12), 1044–1046 (2011).
[CrossRef] [PubMed]

Ha, T.

H. Balci, T. Ha, H. L. Sweeney, and P. R. Selvin, “Interhead distance measurements in myosin VI via SHRImP support a simplified hand-over-hand model,” Biophys. J.89(1), 413–417 (2005).
[CrossRef] [PubMed]

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[CrossRef] [PubMed]

Haeberlé, O.

O. Haeberlé and B. Simon, “Saturated structured confocal microscopy with theoretically unlimited resolution,” Opt. Commun.282(18), 3657–3664 (2009).
[CrossRef]

Han, K. Y.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

Harke, B.

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

Hashimoto, N.

He, J.

S. A. Jones, S. H. Shim, J. He, and X. Zhuang, “Fast, three-dimensional super-resolution imaging of live cells,” Nat. Methods8(6), 499–505 (2011).
[CrossRef] [PubMed]

Heilker, R.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Hell, S. W.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Hess, H. F.

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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Hess, S. T.

S. T. Hess, T. P. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

Hibi, T.

Horanai, H.

Hudmon, A.

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

Jahn, R.

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Jones, S. A.

S. A. Jones, S. H. Shim, J. He, and X. Zhuang, “Fast, three-dimensional super-resolution imaging of live cells,” Nat. Methods8(6), 499–505 (2011).
[CrossRef] [PubMed]

Kasper, R.

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

Keller, J.

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Khonina, S. N.

Kim, D. C.

J. Kim, D. C. Kim, and S. H. Back, “Demonstration of high lateral resolution in laser confocal microscopy using annular and radially polarized light,” Microsc. Res. Tech.72(6), 441–446 (2009).
[CrossRef] [PubMed]

Kim, J.

J. Kim, D. C. Kim, and S. H. Back, “Demonstration of high lateral resolution in laser confocal microscopy using annular and radially polarized light,” Microsc. Res. Tech.72(6), 441–446 (2009).
[CrossRef] [PubMed]

Kner, P.

L. Shao, P. Kner, E. H. Rego, and M. G. Gustafsson, “Super-resolution 3D microscopy of live whole cells using structured illumination,” Nat. Methods8(12), 1044–1046 (2011).
[CrossRef] [PubMed]

Kozawa, Y.

Kredel, S.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Kurihara, M.

Labrakakis, C.

C. Labrakakis, L. E. Lorenzo, C. Bories, A. Ribeiro-da-Silva, and Y. De Koninck, “Inhibitory coupling between inhibitory interneurons in the spinal cord dorsal horn,” Mol. Pain5(1), 24 (2009).
[CrossRef] [PubMed]

Laffray, S.

Lebel, E.

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

Leuchs, G.

R. Dorn, S. Quabis, and G. Leuchs, “Sharper focus for a radially polarized light beam,” Phys. Rev. Lett.91(23), 233901 (2003).
[CrossRef] [PubMed]

Lindwasser, O. W.

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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Lippincott-Schwartz, J.

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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Lorenzo, L. E.

C. Labrakakis, L. E. Lorenzo, C. Bories, A. Ribeiro-da-Silva, and Y. De Koninck, “Inhibitory coupling between inhibitory interneurons in the spinal cord dorsal horn,” Mol. Pain5(1), 24 (2009).
[CrossRef] [PubMed]

Lührmann, R.

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Mason, M. D.

S. T. Hess, T. P. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

McKinney, S. A.

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[CrossRef] [PubMed]

Medda, R.

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Meixner, A. J.

Moneron, G.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

Nemoto, T.

Nienhaus, G. U.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Nienhaus, K.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Olenych, S.

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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Oswald, F.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Patterson, G. H.

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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Pena, A. M.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Piché, M.

Pologruto, T. A.

T. A. Pologruto, B. L. Sabatini, and K. Svoboda, “ScanImage: flexible software for operating laser scanning microscopes,” Biomed. Eng. Online2(1), 13 (2003).
[CrossRef] [PubMed]

Quabis, S.

R. Dorn, S. Quabis, and G. Leuchs, “Sharper focus for a radially polarized light beam,” Phys. Rev. Lett.91(23), 233901 (2003).
[CrossRef] [PubMed]

Rego, E. H.

R. Fiolka, L. Shao, E. H. Rego, M. W. Davidson, and M. G. Gustafsson, “Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination,” Proc. Natl. Acad. Sci. U.S.A.109(14), 5311–5315 (2012).
[CrossRef] [PubMed]

L. Shao, P. Kner, E. H. Rego, and M. G. Gustafsson, “Super-resolution 3D microscopy of live whole cells using structured illumination,” Nat. Methods8(12), 1044–1046 (2011).
[CrossRef] [PubMed]

Reuss, M.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

Ribeiro-da-Silva, A.

C. Labrakakis, L. E. Lorenzo, C. Bories, A. Ribeiro-da-Silva, and Y. De Koninck, “Inhibitory coupling between inhibitory interneurons in the spinal cord dorsal horn,” Mol. Pain5(1), 24 (2009).
[CrossRef] [PubMed]

Rizzoli, S. O.

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Röcker, C.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Roy, H.

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

Rust, M. J.

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

Sabatini, B. L.

T. A. Pologruto, B. L. Sabatini, and K. Svoboda, “ScanImage: flexible software for operating laser scanning microscopes,” Biomed. Eng. Online2(1), 13 (2003).
[CrossRef] [PubMed]

Sato, A.

Sato, S.

Sauer, M.

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

Schanne-Klein, M. C.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Schulman, H.

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

Selvin, P. R.

H. Balci, T. Ha, H. L. Sweeney, and P. R. Selvin, “Interhead distance measurements in myosin VI via SHRImP support a simplified hand-over-hand model,” Biophys. J.89(1), 413–417 (2005).
[CrossRef] [PubMed]

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[CrossRef] [PubMed]

Shao, L.

R. Fiolka, L. Shao, E. H. Rego, M. W. Davidson, and M. G. Gustafsson, “Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination,” Proc. Natl. Acad. Sci. U.S.A.109(14), 5311–5315 (2012).
[CrossRef] [PubMed]

L. Shao, P. Kner, E. H. Rego, and M. G. Gustafsson, “Super-resolution 3D microscopy of live whole cells using structured illumination,” Nat. Methods8(12), 1044–1046 (2011).
[CrossRef] [PubMed]

Shim, S. H.

S. A. Jones, S. H. Shim, J. He, and X. Zhuang, “Fast, three-dimensional super-resolution imaging of live cells,” Nat. Methods8(6), 499–505 (2011).
[CrossRef] [PubMed]

Shroff, H.

H. Shroff, C. G. Galbraith, J. A. Galbraith, and E. Betzig, “Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics,” Nat. Methods5(5), 417–423 (2008).
[CrossRef] [PubMed]

Sik, A.

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

Simon, B.

O. Haeberlé and B. Simon, “Saturated structured confocal microscopy with theoretically unlimited resolution,” Opt. Commun.282(18), 3657–3664 (2009).
[CrossRef]

Sougrat, R.

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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Stadler, J.

Stanciu, C.

Stupperich, C.

Supatto, W.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Svoboda, K.

T. A. Pologruto, B. L. Sabatini, and K. Svoboda, “ScanImage: flexible software for operating laser scanning microscopes,” Biomed. Eng. Online2(1), 13 (2003).
[CrossRef] [PubMed]

Sweeney, H. L.

H. Balci, T. Ha, H. L. Sweeney, and P. R. Selvin, “Interhead distance measurements in myosin VI via SHRImP support a simplified hand-over-hand model,” Biophys. J.89(1), 413–417 (2005).
[CrossRef] [PubMed]

Ta, H.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

Tinnefeld, P.

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

Tordjmann, T.

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

Vallée, R.

Vicidomini, G.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

Waxham, M. N.

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

Westphal, V.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

Wiedenmann, J.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Wolff, M.

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Yildiz, A.

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[CrossRef] [PubMed]

Yokoyama, H.

Zhuang, X.

S. A. Jones, S. H. Shim, J. He, and X. Zhuang, “Fast, three-dimensional super-resolution imaging of live cells,” Nat. Methods8(6), 499–505 (2011).
[CrossRef] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

Appl. Opt. (1)

Biomed. Eng. Online (1)

T. A. Pologruto, B. L. Sabatini, and K. Svoboda, “ScanImage: flexible software for operating laser scanning microscopes,” Biomed. Eng. Online2(1), 13 (2003).
[CrossRef] [PubMed]

Biophys. J. (2)

S. T. Hess, T. P. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J.91(11), 4258–4272 (2006).
[CrossRef] [PubMed]

H. Balci, T. Ha, H. L. Sweeney, and P. R. Selvin, “Interhead distance measurements in myosin VI via SHRImP support a simplified hand-over-hand model,” Biophys. J.89(1), 413–417 (2005).
[CrossRef] [PubMed]

J. Neurosci. (1)

A. Hudmon, E. Lebel, H. Roy, A. Sik, H. Schulman, M. N. Waxham, and P. De Koninck, “A mechanism for Ca2+/calmodulin-dependent protein kinase II clustering at synaptic and nonsynaptic sites based on self-association,” J. Neurosci.25(30), 6971–6983 (2005).
[CrossRef] [PubMed]

J. Opt. Soc. Am. A (1)

Microsc. Res. Tech. (1)

J. Kim, D. C. Kim, and S. H. Back, “Demonstration of high lateral resolution in laser confocal microscopy using annular and radially polarized light,” Microsc. Res. Tech.72(6), 441–446 (2009).
[CrossRef] [PubMed]

Mol. Pain (1)

C. Labrakakis, L. E. Lorenzo, C. Bories, A. Ribeiro-da-Silva, and Y. De Koninck, “Inhibitory coupling between inhibitory interneurons in the spinal cord dorsal horn,” Mol. Pain5(1), 24 (2009).
[CrossRef] [PubMed]

Nat. Methods (6)

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3(10), 793–796 (2006).
[CrossRef] [PubMed]

D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods3(1), 47–53 (2006).
[CrossRef] [PubMed]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, “Sharper low-power STED nanoscopy by time gating,” Nat. Methods8(7), 571–573 (2011).
[CrossRef] [PubMed]

H. Shroff, C. G. Galbraith, J. A. Galbraith, and E. Betzig, “Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics,” Nat. Methods5(5), 417–423 (2008).
[CrossRef] [PubMed]

S. A. Jones, S. H. Shim, J. He, and X. Zhuang, “Fast, three-dimensional super-resolution imaging of live cells,” Nat. Methods8(6), 499–505 (2011).
[CrossRef] [PubMed]

L. Shao, P. Kner, E. H. Rego, and M. G. Gustafsson, “Super-resolution 3D microscopy of live whole cells using structured illumination,” Nat. Methods8(12), 1044–1046 (2011).
[CrossRef] [PubMed]

Opt. Commun. (1)

O. Haeberlé and B. Simon, “Saturated structured confocal microscopy with theoretically unlimited resolution,” Opt. Commun.282(18), 3657–3664 (2009).
[CrossRef]

Opt. Express (2)

Opt. Lett. (2)

Phys. Rev. Lett. (1)

R. Dorn, S. Quabis, and G. Leuchs, “Sharper focus for a radially polarized light beam,” Phys. Rev. Lett.91(23), 233901 (2003).
[CrossRef] [PubMed]

PLoS ONE (1)

S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus, and J. Wiedenmann, “mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures,” PLoS ONE4(2), e4391 (2009).
[CrossRef] [PubMed]

Proc. Natl. Acad. Sci. U.S.A. (2)

R. Fiolka, L. Shao, E. H. Rego, M. W. Davidson, and M. G. Gustafsson, “Time-lapse two-color 3D imaging of live cells with doubled resolution using structured illumination,” Proc. Natl. Acad. Sci. U.S.A.109(14), 5311–5315 (2012).
[CrossRef] [PubMed]

G. Donnert, J. Keller, R. Medda, M. A. Andrei, S. O. Rizzoli, R. Lührmann, R. Jahn, C. Eggeling, and S. W. Hell, “Macromolecular-scale resolution in biological fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.103(31), 11440–11445 (2006).
[CrossRef] [PubMed]

Science (2)

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, “Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization,” Science300(5628), 2061–2065 (2003).
[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,” Science313(5793), 1642–1645 (2006).
[CrossRef] [PubMed]

Small (1)

R. Kasper, B. Harke, C. Forthmann, P. Tinnefeld, S. W. Hell, and M. Sauer, “Single-molecule STED microscopy with photostable organic fluorophores,” Small6(13), 1379–1384 (2010).
[CrossRef] [PubMed]

Other (3)

B. Richards and E. Wolf, “Electromagnetic diffraction in optical systems. 2. Structure of the image field in an aplanatic system,” Proc. R. Soc. A 253, 358–379 (1959).

L. Novotny and B. Hecht, Principles of Nano-Optics, (Cambridge University, 2007).

A. Diaspro, Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances, (Wiley-Liss, 2002).

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