Abstract

Transmissive liquid crystal devices (tLCDs) enable the modification of optical properties, such as phase, polarization, and laser light intensity, over a wide wavelength region at a high conversion efficiency. By utilizing tLCDs, we developed a new two-photon excitation stimulated emission depletion microscopy technique based on a conventional two-photon microscope. Spatial resolution was improved by compensating for phase shifts distributed in the optical path. Using this technique, we observed the fine structures of microtubule networks in fixed biological cells.

© 2014 Optical Society of America

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References

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    [Crossref]
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    [Crossref] [PubMed]

2014 (1)

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

2013 (4)

K. T. Takasaki, J. B. Ding, and B. L. Sabatini, “Live-cell superresolution imaging by pulsed STED two-photon excitation microscopy,” Biophys. J. 104(4), 770–777 (2013).
[Crossref] [PubMed]

P. Bethge, R. Chéreau, E. Avignone, G. Marsicano, and U. V. Nägerl, “Two-photon excitation STED microscopy in two colors in acute brain slices,” Biophys. J. 104(4), 778–785 (2013).
[Crossref] [PubMed]

J. Tønnesen and U. V. Nägerl, “Superresolution imaging for neuroscience,” Exp. Neurol. 242, 33–40 (2013).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

2012 (2)

T. J. Gould, D. Burke, J. Bewersdorf, and M. J. Booth, “Adaptive optics enables 3D STED microscopy in aberrating specimens,” Opt. Express 20(19), 20998–21009 (2012).
[Crossref] [PubMed]

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[Crossref] [PubMed]

2011 (1)

2010 (3)

2009 (3)

2008 (2)

R. K. Singh, P. Senthilkumaran, and K. Singh, “Focusing of linearly, and circularly polarized Gaussian background vortex beams by a high numerical aperture system afflicted with third-order astigmatism,” Opt. Commun. 281(24), 5939–5948 (2008).
[Crossref]

T. Nemoto, “Living cell functions and morphology revealed by two-photon microscopy in intact neural and secretory organs,” Mol. Cells 26(2), 113–120 (2008).
[PubMed]

2007 (1)

S. W. Hell, “Far-field optical nanoscopy,” Science 316(5828), 1153–1158 (2007).
[Crossref] [PubMed]

2006 (2)

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]

2005 (1)

M. G. L. Gustafsson, “Nonlinear structured-illumination microscopy: wide-Field fluorescence imaging with theoretically unlimited resolution,” Proc. Natl. Acad. Sci. U.S.A. 102(37), 13081–13086 (2005).
[Crossref] [PubMed]

2003 (1)

S. W. Hell, “Toward fluorescence nanoscopy,” Nat. Biotechnol. 21(11), 1347–1355 (2003).
[Crossref] [PubMed]

1994 (1)

1959 (1)

B. Richards and E. Wolf, “Electromagnetic diffraction in optical system II. Structure of the image field in an aplanatic system,” Proc. R. Soc. Lond. A Math. Phys. Sci. 253(1274), 358–379 (1959).
[Crossref]

Artal, P.

Avignone, E.

P. Bethge, R. Chéreau, E. Avignone, G. Marsicano, and U. V. Nägerl, “Two-photon excitation STED microscopy in two colors in acute brain slices,” Biophys. J. 104(4), 778–785 (2013).
[Crossref] [PubMed]

Bates, M.

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]

Bethge, P.

P. Bethge, R. Chéreau, E. Avignone, G. Marsicano, and U. V. Nägerl, “Two-photon excitation STED microscopy in two colors in acute brain slices,” Biophys. J. 104(4), 778–785 (2013).
[Crossref] [PubMed]

Betzig, E.

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]

Bewersdorf, J.

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,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Booth, M. J.

Burke, D.

Cheng, Y.

Chéreau, R.

P. Bethge, R. Chéreau, E. Avignone, G. Marsicano, and U. V. Nägerl, “Two-photon excitation STED microscopy in two colors in acute brain slices,” Biophys. J. 104(4), 778–785 (2013).
[Crossref] [PubMed]

Chou, K. C.

Q. Li, S. S. Wu, and K. C. Chou, “Subdiffraction-limit two-photon fluorescence microscopy for GFP-tagged cell imaging,” Biophys. J. 97(12), 3224–3228 (2009).
[Crossref] [PubMed]

Davidson, M. 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,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Deng, S.

Ding, J. B.

K. T. Takasaki, J. B. Ding, and B. L. Sabatini, “Live-cell superresolution imaging by pulsed STED two-photon excitation microscopy,” Biophys. J. 104(4), 770–777 (2013).
[Crossref] [PubMed]

Fernández, E. J.

Gould, T. J.

Gustafsson, M. G. L.

M. G. L. Gustafsson, “Nonlinear structured-illumination microscopy: wide-Field fluorescence imaging with theoretically unlimited resolution,” Proc. Natl. Acad. Sci. U.S.A. 102(37), 13081–13086 (2005).
[Crossref] [PubMed]

Hall, S.

Hao, X.

X. Hao, C. Kuang, T. Wang, and X. Liu, “Effects of polarization on the de-excitation dark focal spot in STED microscopy,” J. Opt. 12(11), 115707 (2010).
[Crossref]

Hashimoto, N.

Hell, S. W.

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,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Hibi, T.

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

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. Express 19(17), 15947–15954 (2011).
[Crossref] [PubMed]

Horanai, H.

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

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. Express 19(17), 15947–15954 (2011).
[Crossref] [PubMed]

Ipponjima, S.

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

Kasai, H.

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[Crossref] [PubMed]

Kawakami, R.

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

Knox, S.

Kozawa, Y.

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

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. Express 19(17), 15947–15954 (2011).
[Crossref] [PubMed]

Kuang, C.

X. Hao, C. Kuang, T. Wang, and X. Liu, “Effects of polarization on the de-excitation dark focal spot in STED microscopy,” J. Opt. 12(11), 115707 (2010).
[Crossref]

Kurihara, M.

Li, Q.

Q. Li, S. S. Wu, and K. C. Chou, “Subdiffraction-limit two-photon fluorescence microscopy for GFP-tagged cell imaging,” Biophys. J. 97(12), 3224–3228 (2009).
[Crossref] [PubMed]

Li, R.

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,” Science 313(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,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Liu, L.

Liu, X.

X. Hao, C. Kuang, T. Wang, and X. Liu, “Effects of polarization on the de-excitation dark focal spot in STED microscopy,” J. Opt. 12(11), 115707 (2010).
[Crossref]

Marsicano, G.

P. Bethge, R. Chéreau, E. Avignone, G. Marsicano, and U. V. Nägerl, “Two-photon excitation STED microscopy in two colors in acute brain slices,” Biophys. J. 104(4), 778–785 (2013).
[Crossref] [PubMed]

Miki, T.

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[Crossref] [PubMed]

Moneron, G.

Nägerl, U. V.

P. Bethge, R. Chéreau, E. Avignone, G. Marsicano, and U. V. Nägerl, “Two-photon excitation STED microscopy in two colors in acute brain slices,” Biophys. J. 104(4), 778–785 (2013).
[Crossref] [PubMed]

J. Tønnesen and U. V. Nägerl, “Superresolution imaging for neuroscience,” Exp. Neurol. 242, 33–40 (2013).
[Crossref] [PubMed]

Nemoto, T.

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[Crossref] [PubMed]

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. Express 19(17), 15947–15954 (2011).
[Crossref] [PubMed]

T. Nemoto, “Living cell functions and morphology revealed by two-photon microscopy in intact neural and secretory organs,” Mol. Cells 26(2), 113–120 (2008).
[PubMed]

Ogata, S.

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[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,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Paterson, C.

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,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Prieto, P. M.

Richards, B.

B. Richards and E. Wolf, “Electromagnetic diffraction in optical system II. Structure of the image field in an aplanatic system,” Proc. R. Soc. Lond. A Math. Phys. Sci. 253(1274), 358–379 (1959).
[Crossref]

Rust, M. J.

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]

Sabatini, B. L.

K. T. Takasaki, J. B. Ding, and B. L. Sabatini, “Live-cell superresolution imaging by pulsed STED two-photon excitation microscopy,” Biophys. J. 104(4), 770–777 (2013).
[Crossref] [PubMed]

Sato, A.

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

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. Express 19(17), 15947–15954 (2011).
[Crossref] [PubMed]

Sato, S.

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

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. Express 19(17), 15947–15954 (2011).
[Crossref] [PubMed]

Sawada, K.

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

Seino, S.

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[Crossref] [PubMed]

Senthilkumaran, P.

R. K. Singh, P. Senthilkumaran, and K. Singh, “Focusing of linearly, and circularly polarized Gaussian background vortex beams by a high numerical aperture system afflicted with third-order astigmatism,” Opt. Commun. 281(24), 5939–5948 (2008).
[Crossref]

Shaw, M.

Singh, K.

R. K. Singh, P. Senthilkumaran, and K. Singh, “Focusing of linearly, and circularly polarized Gaussian background vortex beams by a high numerical aperture system afflicted with third-order astigmatism,” Opt. Commun. 281(24), 5939–5948 (2008).
[Crossref]

Singh, R. K.

R. K. Singh, P. Senthilkumaran, and K. Singh, “Focusing of linearly, and circularly polarized Gaussian background vortex beams by a high numerical aperture system afflicted with third-order astigmatism,” Opt. Commun. 281(24), 5939–5948 (2008).
[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,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Stevens, R.

Takasaki, K. T.

K. T. Takasaki, J. B. Ding, and B. L. Sabatini, “Live-cell superresolution imaging by pulsed STED two-photon excitation microscopy,” Biophys. J. 104(4), 770–777 (2013).
[Crossref] [PubMed]

Tamai, S.

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[Crossref] [PubMed]

Tønnesen, J.

J. Tønnesen and U. V. Nägerl, “Superresolution imaging for neuroscience,” Exp. Neurol. 242, 33–40 (2013).
[Crossref] [PubMed]

Wang, T.

X. Hao, C. Kuang, T. Wang, and X. Liu, “Effects of polarization on the de-excitation dark focal spot in STED microscopy,” J. Opt. 12(11), 115707 (2010).
[Crossref]

Wichmann, J.

Wolf, E.

B. Richards and E. Wolf, “Electromagnetic diffraction in optical system II. Structure of the image field in an aplanatic system,” Proc. R. Soc. Lond. A Math. Phys. Sci. 253(1274), 358–379 (1959).
[Crossref]

Wu, S. S.

Q. Li, S. S. Wu, and K. C. Chou, “Subdiffraction-limit two-photon fluorescence microscopy for GFP-tagged cell imaging,” Biophys. J. 97(12), 3224–3228 (2009).
[Crossref] [PubMed]

Xu, Z.

Yokoyama, H.

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

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. Express 19(17), 15947–15954 (2011).
[Crossref] [PubMed]

Zhuang, X.

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]

Biophys. J. (3)

Q. Li, S. S. Wu, and K. C. Chou, “Subdiffraction-limit two-photon fluorescence microscopy for GFP-tagged cell imaging,” Biophys. J. 97(12), 3224–3228 (2009).
[Crossref] [PubMed]

K. T. Takasaki, J. B. Ding, and B. L. Sabatini, “Live-cell superresolution imaging by pulsed STED two-photon excitation microscopy,” Biophys. J. 104(4), 770–777 (2013).
[Crossref] [PubMed]

P. Bethge, R. Chéreau, E. Avignone, G. Marsicano, and U. V. Nägerl, “Two-photon excitation STED microscopy in two colors in acute brain slices,” Biophys. J. 104(4), 778–785 (2013).
[Crossref] [PubMed]

Exp. Neurol. (1)

J. Tønnesen and U. V. Nägerl, “Superresolution imaging for neuroscience,” Exp. Neurol. 242, 33–40 (2013).
[Crossref] [PubMed]

J. Opt. (1)

X. Hao, C. Kuang, T. Wang, and X. Liu, “Effects of polarization on the de-excitation dark focal spot in STED microscopy,” J. Opt. 12(11), 115707 (2010).
[Crossref]

Microscopy (Oxf) (1)

S. Ipponjima, T. Hibi, Y. Kozawa, H. Horanai, H. Yokoyama, S. Sato, and T. Nemoto, “Improvement of lateral resolution and extension of depth of field in two-photon microscopy by a higher-order radially polarized beam,” Microscopy (Oxf) 63(1), 23–32 (2014).
[Crossref] [PubMed]

Mol. Cells (1)

T. Nemoto, “Living cell functions and morphology revealed by two-photon microscopy in intact neural and secretory organs,” Mol. Cells 26(2), 113–120 (2008).
[PubMed]

Nat. Biotechnol. (1)

S. W. Hell, “Toward fluorescence nanoscopy,” Nat. Biotechnol. 21(11), 1347–1355 (2003).
[Crossref] [PubMed]

Nat. Methods (1)

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]

Opt. Commun. (1)

R. K. Singh, P. Senthilkumaran, and K. Singh, “Focusing of linearly, and circularly polarized Gaussian background vortex beams by a high numerical aperture system afflicted with third-order astigmatism,” Opt. Commun. 281(24), 5939–5948 (2008).
[Crossref]

Opt. Express (6)

Opt. Lett. (1)

PLoS ONE (1)

S. Ogata, T. Miki, S. Seino, S. Tamai, H. Kasai, and T. Nemoto, “A novel function of Noc2 in agonist-induced intracellular Ca2+ increase during zymogen-granule exocytosis in pancreatic acinar cells,” PLoS ONE 7(5), e37048 (2012).
[Crossref] [PubMed]

Proc. Natl. Acad. Sci. U.S.A. (1)

M. G. L. Gustafsson, “Nonlinear structured-illumination microscopy: wide-Field fluorescence imaging with theoretically unlimited resolution,” Proc. Natl. Acad. Sci. U.S.A. 102(37), 13081–13086 (2005).
[Crossref] [PubMed]

Proc. R. Soc. Lond. A Math. Phys. Sci. (1)

B. Richards and E. Wolf, “Electromagnetic diffraction in optical system II. Structure of the image field in an aplanatic system,” Proc. R. Soc. Lond. A Math. Phys. Sci. 253(1274), 358–379 (1959).
[Crossref]

Sci Rep (1)

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci Rep 3, 1014 (2013).
[Crossref] [PubMed]

Science (2)

S. W. Hell, “Far-field optical nanoscopy,” Science 316(5828), 1153–1158 (2007).
[Crossref] [PubMed]

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

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Figures (4)

Fig. 1
Fig. 1

(a) Schematic of our TP-STED system using tLCDs. DM: dichroic mirror; F: filter; GM: galvano mirrors; tLCD: transmissive liquid crystal device; -P: plain cell; −24: 24-divided cell; M: mirror; OL: objective lens; PMT: photomultiplier tube; TL: tube lenses; λ/2: half-wave plate. (b) Theoretical phase distribution of an optical vortex generated by tLCD-24.

Fig. 2
Fig. 2

(a) Calculated intensity distribution in the focal plane for focusing of the circularly polarized 577 nm optical vortex created by tLCD-24. (b)–(d) Fluorescence images of a fluorescent bead directly excited by optical vortexes. The lower panels show the fluorescence intensity profiles across the intensity center along the x-axis. (b) Image of a 170 nm orange bead excited by 577 nm light with circular polarization at the objective lens. (c) Image of a 170 nm orange bead excited by 577 nm light with circular polarization at tLCDs. (d) Image of a 170 nm yellow-green bead excited by 473 nm light with circular polarization at the objective lens.

Fig. 3
Fig. 3

Merged fluorescence image of 100 nm green beads (green) and 170 nm orange beads (magenta) placed on the same cover slip.

Fig. 4
Fig. 4

Comparisons of TPLSM and TP-STED images. (a) A 100 nm green fluorescent bead. The lower panels show the averaged fluorescence intensity profiles of n beads across the intensity center along the red dashed lines in the fluorescent images. Inlet length value indicates the full width at half maximum. (b) Microtubule networks in fixed COS-7 cells after immunostaining with antibodies conjugated with the fluorescent dye ATTO 425. The lower panels show the fluorescence intensity profiles across the red dashed lines in the fluorescent images.

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