Abstract

One of the main challenges in three-dimensional microscopy is to overcome the lack of isotropy of the spatial resolution, which results from the axially-elongated shape of the point spread function. Such anisotropy gives rise to images in which significant axially-oriented structures of the sample are not resolved. In this paper we achieve an important improvement in z resolution in two-photon excitation microscopy through spatial modulation of the incident beam. Specifically, we demonstrate that the design and implementation of a simple shaded ring performs quasi-isotropic three-dimensional imaging and that the corresponding loss in luminosity can be easily compensated by most available femtosecond lasers. The outcome looks particularly relevant to nano-fabrication and optical manipulation.

© 2005 Optical Society of America

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  1. E. Abbe,Arch. Mikrosk. Anat. 9,413–468 (1873).
    [CrossRef]
  2. T. R. M. Sales, “Smallest focal spot,” Phys. Rev. Lett. 81, 3844–3847 (1998).
    [CrossRef]
  3. J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
    [CrossRef] [PubMed]
  4. J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
    [CrossRef] [PubMed]
  5. C. M. Blanca and S. W. Hell, “Axial superresolution with ultrahigh aperture lenses,” Opt. Express 10, 893–898 (2002).
    [PubMed]
  6. J. B. Pawley, ed., Handbook of biological confocal microscopy. Plenum Press, New York,1995.
  7. The term superresolution introduced here is understood in the sense of Rayleigh criterion; i. e., as the narrowness of the PSF or, equivalently, the enhancement of the OTF for frequencies under the cut-off frequency.
  8. S. Lindek, J. Swoger, and E. H. K. Stelzer, “Single-lens theta microscopy: resolution, efficiency and working distance,” J. Mod. Opt. 46, 843–858 (1999).
  9. B. Bailey, D. L. Farkas, D. Lansing-Taylor, and F. Lanni, “Enhancement resolution in fluorescence microscopy by standing-wave excitation,” Science 366, 44–48 (1993).
  10. J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
    [CrossRef] [PubMed]
  11. M. A. A. Neil, R. Juskaitis, T. Wilson, Z. J. Laczik, and V. Sarafis, “Optimized pupil-plane filters for confo-cal microscope point-spread function engineering,”Opt. Lett. 25, 245–247 (2000).
    [CrossRef]
  12. C. J. R. Sheppard, “Binary optics and confocal imaging,” Opt. Lett. 24, 505–506 (1999).
    [CrossRef]
  13. M. Martínez-Corral, M. T. Caballero, E. H. K. Stelzer, and J. Swoger, 2201C;Tailoring the axial shape of the point spread function using the Toraldo concept,” Opt. Express 10, 98–103 (2002).
    [PubMed]
  14. G. Boyer, “New class of axially apodizing filters for confocal scanning microscopy,” J. Opt. Soc. Am. A 19, 584–589 (2002).
    [CrossRef]
  15. M. Martínez-Corral, C. IbáÑez-López, G. Saavedra, and M. T. Caballero, “Axial gain in resolution in optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 11, 1740–1745 (2003).
    [CrossRef] [PubMed]
  16. S. S. Sherif and P. Török, “Pupil plane masks for super-resolution in high-numerical-aperture focusing,” J. Mod. Opt. 51, 2007–2019 (2004).
  17. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
    [CrossRef] [PubMed]
  18. C. M. Blanca, J. Bewersdorf, and S. W. Hell, “Single sharp spot in fluorescence microscopy of two opposing lenses,” Appl. Phys. Lett. 79, 2321–2323 (2001).
    [CrossRef]
  19. T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
    [CrossRef]
  20. M. Dyba and S. W. Hell, “Focal spots of size lambda/23 open up far-field fluorescence microscopy at 33 nm axial resolution,” Phys. Rev. Lett. 88, 163901 (2002).
    [CrossRef] [PubMed]
  21. C. IbáÑez-López, I. Escobar, G. Saavedra, and M. Martínez-Corral, “Optical sectioning improvement in two-color excitation scanning microscopy,” Microsc. Res. Tech. 64, 96–102 (2004).
    [CrossRef] [PubMed]
  22. The eccentricity is defined here as e=1−b2a2,, where a and b account for the lengths of the semimajor and semiminor axes, respectively.
  23. S. Kawata, H.-B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697–698 (2001).
    [CrossRef] [PubMed]
  24. D. G. Grier, “A revolution in optical manipulation,” Nature 424, 810–816 (2003).
    [CrossRef] [PubMed]
  25. B. Richards and E. Wolf, Proceedings of the Royal Society (London) A253, 358 (1959).
    [CrossRef]
  26. P. Török and P. Varga, “Electromagnetic diffraction of light focused through a stratified medium,” Appl. Opt. 36, 2305–2312 (1997)
    [CrossRef] [PubMed]
  27. O. Haeberlé, “Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part I: Conventional microscopy,” Opt. Commun. 216, 55–63 (2003).
    [CrossRef]

2004 (3)

J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
[CrossRef] [PubMed]

S. S. Sherif and P. Török, “Pupil plane masks for super-resolution in high-numerical-aperture focusing,” J. Mod. Opt. 51, 2007–2019 (2004).

C. IbáÑez-López, I. Escobar, G. Saavedra, and M. Martínez-Corral, “Optical sectioning improvement in two-color excitation scanning microscopy,” Microsc. Res. Tech. 64, 96–102 (2004).
[CrossRef] [PubMed]

2003 (3)

D. G. Grier, “A revolution in optical manipulation,” Nature 424, 810–816 (2003).
[CrossRef] [PubMed]

O. Haeberlé, “Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part I: Conventional microscopy,” Opt. Commun. 216, 55–63 (2003).
[CrossRef]

M. Martínez-Corral, C. IbáÑez-López, G. Saavedra, and M. T. Caballero, “Axial gain in resolution in optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 11, 1740–1745 (2003).
[CrossRef] [PubMed]

2002 (6)

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
[CrossRef] [PubMed]

M. Martínez-Corral, M. T. Caballero, E. H. K. Stelzer, and J. Swoger, 2201C;Tailoring the axial shape of the point spread function using the Toraldo concept,” Opt. Express 10, 98–103 (2002).
[PubMed]

G. Boyer, “New class of axially apodizing filters for confocal scanning microscopy,” J. Opt. Soc. Am. A 19, 584–589 (2002).
[CrossRef]

C. M. Blanca and S. W. Hell, “Axial superresolution with ultrahigh aperture lenses,” Opt. Express 10, 893–898 (2002).
[PubMed]

M. Dyba and S. W. Hell, “Focal spots of size lambda/23 open up far-field fluorescence microscopy at 33 nm axial resolution,” Phys. Rev. Lett. 88, 163901 (2002).
[CrossRef] [PubMed]

2001 (2)

S. Kawata, H.-B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697–698 (2001).
[CrossRef] [PubMed]

C. M. Blanca, J. Bewersdorf, and S. W. Hell, “Single sharp spot in fluorescence microscopy of two opposing lenses,” Appl. Phys. Lett. 79, 2321–2323 (2001).
[CrossRef]

2000 (2)

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
[CrossRef]

M. A. A. Neil, R. Juskaitis, T. Wilson, Z. J. Laczik, and V. Sarafis, “Optimized pupil-plane filters for confo-cal microscope point-spread function engineering,”Opt. Lett. 25, 245–247 (2000).
[CrossRef]

1999 (2)

C. J. R. Sheppard, “Binary optics and confocal imaging,” Opt. Lett. 24, 505–506 (1999).
[CrossRef]

S. Lindek, J. Swoger, and E. H. K. Stelzer, “Single-lens theta microscopy: resolution, efficiency and working distance,” J. Mod. Opt. 46, 843–858 (1999).

1998 (1)

T. R. M. Sales, “Smallest focal spot,” Phys. Rev. Lett. 81, 3844–3847 (1998).
[CrossRef]

1997 (1)

1993 (1)

B. Bailey, D. L. Farkas, D. Lansing-Taylor, and F. Lanni, “Enhancement resolution in fluorescence microscopy by standing-wave excitation,” Science 366, 44–48 (1993).

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

1873 (1)

E. Abbe,Arch. Mikrosk. Anat. 9,413–468 (1873).
[CrossRef]

Abbe, E.

E. Abbe,Arch. Mikrosk. Anat. 9,413–468 (1873).
[CrossRef]

Anderson, E. K.

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

Bailey, B.

B. Bailey, D. L. Farkas, D. Lansing-Taylor, and F. Lanni, “Enhancement resolution in fluorescence microscopy by standing-wave excitation,” Science 366, 44–48 (1993).

Bene, F.Del

J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
[CrossRef] [PubMed]

Bewersdorf, J.

C. M. Blanca, J. Bewersdorf, and S. W. Hell, “Single sharp spot in fluorescence microscopy of two opposing lenses,” Appl. Phys. Lett. 79, 2321–2323 (2001).
[CrossRef]

Blanca, C. M.

C. M. Blanca and S. W. Hell, “Axial superresolution with ultrahigh aperture lenses,” Opt. Express 10, 893–898 (2002).
[PubMed]

C. M. Blanca, J. Bewersdorf, and S. W. Hell, “Single sharp spot in fluorescence microscopy of two opposing lenses,” Appl. Phys. Lett. 79, 2321–2323 (2001).
[CrossRef]

Boyer, G.

Caballero, M. T.

Denk, W.

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

Dyba, M.

M. Dyba and S. W. Hell, “Focal spots of size lambda/23 open up far-field fluorescence microscopy at 33 nm axial resolution,” Phys. Rev. Lett. 88, 163901 (2002).
[CrossRef] [PubMed]

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
[CrossRef]

Egner, A.

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
[CrossRef]

Escobar, I.

C. IbáÑez-López, I. Escobar, G. Saavedra, and M. Martínez-Corral, “Optical sectioning improvement in two-color excitation scanning microscopy,” Microsc. Res. Tech. 64, 96–102 (2004).
[CrossRef] [PubMed]

Farkas, D. L.

B. Bailey, D. L. Farkas, D. Lansing-Taylor, and F. Lanni, “Enhancement resolution in fluorescence microscopy by standing-wave excitation,” Science 366, 44–48 (1993).

Grier, D. G.

D. G. Grier, “A revolution in optical manipulation,” Nature 424, 810–816 (2003).
[CrossRef] [PubMed]

Haeberlé, O.

O. Haeberlé, “Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part I: Conventional microscopy,” Opt. Commun. 216, 55–63 (2003).
[CrossRef]

Hell, S. W.

C. M. Blanca and S. W. Hell, “Axial superresolution with ultrahigh aperture lenses,” Opt. Express 10, 893–898 (2002).
[PubMed]

M. Dyba and S. W. Hell, “Focal spots of size lambda/23 open up far-field fluorescence microscopy at 33 nm axial resolution,” Phys. Rev. Lett. 88, 163901 (2002).
[CrossRef] [PubMed]

C. M. Blanca, J. Bewersdorf, and S. W. Hell, “Single sharp spot in fluorescence microscopy of two opposing lenses,” Appl. Phys. Lett. 79, 2321–2323 (2001).
[CrossRef]

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
[CrossRef]

Hodgson, K. O.

J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
[CrossRef] [PubMed]

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

Huisken, J.

J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
[CrossRef] [PubMed]

IbáÑez-López, C.

C. IbáÑez-López, I. Escobar, G. Saavedra, and M. Martínez-Corral, “Optical sectioning improvement in two-color excitation scanning microscopy,” Microsc. Res. Tech. 64, 96–102 (2004).
[CrossRef] [PubMed]

M. Martínez-Corral, C. IbáÑez-López, G. Saavedra, and M. T. Caballero, “Axial gain in resolution in optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 11, 1740–1745 (2003).
[CrossRef] [PubMed]

Ishikawa, T.

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

Jakobs, S.

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
[CrossRef]

Johnson, B.

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

Juskaitis, R.

Kawata, S.

S. Kawata, H.-B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697–698 (2001).
[CrossRef] [PubMed]

Klar, T. A.

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
[CrossRef]

Laczik, Z. J.

Lai, B.

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

Lanni, F.

B. Bailey, D. L. Farkas, D. Lansing-Taylor, and F. Lanni, “Enhancement resolution in fluorescence microscopy by standing-wave excitation,” Science 366, 44–48 (1993).

Lansing-Taylor, D.

B. Bailey, D. L. Farkas, D. Lansing-Taylor, and F. Lanni, “Enhancement resolution in fluorescence microscopy by standing-wave excitation,” Science 366, 44–48 (1993).

Lindek, S.

S. Lindek, J. Swoger, and E. H. K. Stelzer, “Single-lens theta microscopy: resolution, efficiency and working distance,” J. Mod. Opt. 46, 843–858 (1999).

Martínez-Corral, M.

Miao, J.

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
[CrossRef] [PubMed]

Neil, M. A. A.

O’Keefe, M. A.

J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
[CrossRef] [PubMed]

Ohsuna, T.

J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
[CrossRef] [PubMed]

Richards, B.

B. Richards and E. Wolf, Proceedings of the Royal Society (London) A253, 358 (1959).
[CrossRef]

Saavedra, G.

C. IbáÑez-López, I. Escobar, G. Saavedra, and M. Martínez-Corral, “Optical sectioning improvement in two-color excitation scanning microscopy,” Microsc. Res. Tech. 64, 96–102 (2004).
[CrossRef] [PubMed]

M. Martínez-Corral, C. IbáÑez-López, G. Saavedra, and M. T. Caballero, “Axial gain in resolution in optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 11, 1740–1745 (2003).
[CrossRef] [PubMed]

Sales, T. R. M.

T. R. M. Sales, “Smallest focal spot,” Phys. Rev. Lett. 81, 3844–3847 (1998).
[CrossRef]

Sarafis, V.

Sheppard, C. J. R.

Sherif, S. S.

S. S. Sherif and P. Török, “Pupil plane masks for super-resolution in high-numerical-aperture focusing,” J. Mod. Opt. 51, 2007–2019 (2004).

Stelzer, E. H. K.

J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
[CrossRef] [PubMed]

M. Martínez-Corral, M. T. Caballero, E. H. K. Stelzer, and J. Swoger, 2201C;Tailoring the axial shape of the point spread function using the Toraldo concept,” Opt. Express 10, 98–103 (2002).
[PubMed]

S. Lindek, J. Swoger, and E. H. K. Stelzer, “Single-lens theta microscopy: resolution, efficiency and working distance,” J. Mod. Opt. 46, 843–858 (1999).

Strickler, J. H.

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

Sun, H.-B.

S. Kawata, H.-B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697–698 (2001).
[CrossRef] [PubMed]

Swoger, J.

J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
[CrossRef] [PubMed]

M. Martínez-Corral, M. T. Caballero, E. H. K. Stelzer, and J. Swoger, 2201C;Tailoring the axial shape of the point spread function using the Toraldo concept,” Opt. Express 10, 98–103 (2002).
[PubMed]

S. Lindek, J. Swoger, and E. H. K. Stelzer, “Single-lens theta microscopy: resolution, efficiency and working distance,” J. Mod. Opt. 46, 843–858 (1999).

Takada, K.

S. Kawata, H.-B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697–698 (2001).
[CrossRef] [PubMed]

Tanaka, T.

S. Kawata, H.-B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697–698 (2001).
[CrossRef] [PubMed]

Terasaki, O.

J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
[CrossRef] [PubMed]

Török, P.

S. S. Sherif and P. Török, “Pupil plane masks for super-resolution in high-numerical-aperture focusing,” J. Mod. Opt. 51, 2007–2019 (2004).

P. Török and P. Varga, “Electromagnetic diffraction of light focused through a stratified medium,” Appl. Opt. 36, 2305–2312 (1997)
[CrossRef] [PubMed]

Varga, P.

Webb, W. W.

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

Wilson, T.

Wittbrodt, J.

J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
[CrossRef] [PubMed]

Wolf, E.

B. Richards and E. Wolf, Proceedings of the Royal Society (London) A253, 358 (1959).
[CrossRef]

Appl. Opt. (1)

Appl. Phys. Lett. (1)

C. M. Blanca, J. Bewersdorf, and S. W. Hell, “Single sharp spot in fluorescence microscopy of two opposing lenses,” Appl. Phys. Lett. 79, 2321–2323 (2001).
[CrossRef]

Arch. Mikrosk. Anat. (1)

E. Abbe,Arch. Mikrosk. Anat. 9,413–468 (1873).
[CrossRef]

J. Mod. Opt. (2)

S. S. Sherif and P. Török, “Pupil plane masks for super-resolution in high-numerical-aperture focusing,” J. Mod. Opt. 51, 2007–2019 (2004).

S. Lindek, J. Swoger, and E. H. K. Stelzer, “Single-lens theta microscopy: resolution, efficiency and working distance,” J. Mod. Opt. 46, 843–858 (1999).

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

Microsc. Res. Tech. (1)

C. IbáÑez-López, I. Escobar, G. Saavedra, and M. Martínez-Corral, “Optical sectioning improvement in two-color excitation scanning microscopy,” Microsc. Res. Tech. 64, 96–102 (2004).
[CrossRef] [PubMed]

Nature (2)

S. Kawata, H.-B. Sun, T. Tanaka, and K. Takada, “Finer features for functional microdevices,” Nature 412, 697–698 (2001).
[CrossRef] [PubMed]

D. G. Grier, “A revolution in optical manipulation,” Nature 424, 810–816 (2003).
[CrossRef] [PubMed]

Opt. Commun. (1)

O. Haeberlé, “Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part I: Conventional microscopy,” Opt. Commun. 216, 55–63 (2003).
[CrossRef]

Opt. Express (3)

Opt. Lett. (2)

Phys. Rev. Lett. (4)

M. Dyba and S. W. Hell, “Focal spots of size lambda/23 open up far-field fluorescence microscopy at 33 nm axial resolution,” Phys. Rev. Lett. 88, 163901 (2002).
[CrossRef] [PubMed]

T. R. M. Sales, “Smallest focal spot,” Phys. Rev. Lett. 81, 3844–3847 (1998).
[CrossRef]

J. Miao, T. Ishikawa, B. Johnson, E. K. Anderson, B. Lai, and K. O. Hodgson, “High resolution 3D x-ray diffraction microscopy,” Phys. Rev. Lett. 89, 088303 (2002).
[CrossRef] [PubMed]

J. Miao, T. Ohsuna, O. Terasaki, K. O. Hodgson, and M. A. O’Keefe, “Atomic resolution three-dimensional electron diffraction microscopy,” Phys. Rev. Lett. 89, 155502 (2002).
[CrossRef] [PubMed]

Proc. Natl. Acad. Sc. (1)

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sc. 97, 8206–8210 (2000).
[CrossRef]

Science (3)

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

B. Bailey, D. L. Farkas, D. Lansing-Taylor, and F. Lanni, “Enhancement resolution in fluorescence microscopy by standing-wave excitation,” Science 366, 44–48 (1993).

J. Huisken, J. Swoger, F.Del Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,”Science 305, 1007–1009 (2004).
[CrossRef] [PubMed]

Other (4)

J. B. Pawley, ed., Handbook of biological confocal microscopy. Plenum Press, New York,1995.

The term superresolution introduced here is understood in the sense of Rayleigh criterion; i. e., as the narrowness of the PSF or, equivalently, the enhancement of the OTF for frequencies under the cut-off frequency.

B. Richards and E. Wolf, Proceedings of the Royal Society (London) A253, 358 (1959).
[CrossRef]

The eccentricity is defined here as e=1−b2a2,, where a and b account for the lengths of the semimajor and semiminor axes, respectively.

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Figures (5)

Fig. 1.
Fig. 1.

Amplitude transmittance of the SR filter. The transmittance of the shaded ring is 19%.

Fig. 2.
Fig. 2.

Plot of the numerically evaluated 3-D PSF, in the meridian plan φ = 0 , corresponding to: (a) Standard 1.2 NA objective with clear pupil; and (b) Same objective but with the SR filter inserted as the aperture stop. Axes labels are expressed in microns.

Fig. 3.
Fig. 3.

-Schematic geometry of the TPE scanning microscope.

Fig. 4.
Fig. 4.

- 2D gray-scale sections of the experimental 3-D PSFs as displayed by LabView. (a) PSF obtained with the standard 1.2 NA objective with clear pupil; and (b) PSF obtained with the same objective but with the SR filter inserted as the aperture stop.

Fig. 5.
Fig. 5.

- Normalized axial and lateral PSF produced by the Olympus 1.2 NA objective (x). With empty dots (o) we show the experimental results obtained when the SR filter is inserted in the illumination path.

Equations (5)

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E ( r , z , φ ) = [ I 0 ( r , z ) + I 2 ( r , z ) cos 2 φ ] i + I 2 ( r , z ) sin 2 φ j + i 2 I 1 ( r , z ) cos φ k
I 0 ( r , z ) = 0 α P ( θ ) cos θ ( 1 + cos θ ) J 0 ( k r sin θ ) exp ( ik z cos θ ) sin θ d θ
I 1 ( r , z ) = 0 α P ( θ ) cos θ sin θ J 1 ( k r sin θ ) exp ( ik z cos θ ) sin θ d θ
I 2 ( r , z ) = 0 α P ( θ ) cos θ ( 1 cos θ ) J 2 ( k r sin θ ) exp ( ik z cos θ ) sin θ d θ
PSF ( r , z , φ ) = E ( r , z , φ ) 4 .

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