Abstract

Structured illumination has been employed in fluorescence microscopy to extend its lateral resolution. It has been demonstrated that a factor of 2 improvement can be achieved. In this paper, we introduce a novel optical arrangement that can further improve the resolution. It makes use of a fine grating held in close proximity to the sample. The fringe pattern thus projected onto the sample contains grating vectors substantially higher than those that are possible with the conventional structured illumination setup. We will present experimental results to demonstrate the principle of the technique, and will show that, theoretically, it can achieve an imaging NA approaching 4.

© 2010 Optical Society of America

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  1. R. Heintzmann and C. Cremer, “Laterally modulated excitation microscopy: Improvement of resolution by using a diffraction grating,” Proc. SPIE 3568, 185–196 (1999).
    [CrossRef]
  2. M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc. 198, 82–87 (2000).
    [CrossRef]
  3. S. W. Hell and Jan Wichmann, “Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy,” Opt. Lett. 19, 780–782(1994).
    [CrossRef]
  4. S. W. Hell, E. H. K. Stelzer, S. Lindek, and C. Cremer, “Confocal microscopy with an increased detection aperture: type-B 4Pi confocal microscopy,” Opt. Lett. 19, 222–224 (1994).
    [CrossRef]
  5. M. G. L. Gustafsson, D. A. Agard, and J. W. Sedat, “I5M: 3D widefield light microscopy with better than 100nm axial resolution,” J. Microsc. 195, 10–16 (1999).
    [CrossRef]
  6. B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, “Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation,” Nature 366, 44–48 (1993).
    [CrossRef]
  7. M. G. Somekh, K. Hsu, and M. C. Pitter, “Resolution in structured illumination microscopy: a probabilistic approach,” J. Opt. Soc. Am. 25, 1319–1329 (2008).
    [CrossRef]
  8. S. Liu, C. J. Chuang, C. W. See, G. Zoriniants, W. L. Barnes, and M. G. Somekh, “Double-grating-structured light microscope using plasmonic nanoparticle arrays,” Opt. Lett. 34, 1255–1257 (2009).
    [CrossRef]
  9. The value of the k-vector is normalized with respect to 2π/λ so that it has a maximum value of 1 in air.
  10. A. Sentenac, K. Belkebir, H. Giovannini, and P. C. Chaumet, “Subdiffraction resolution in total internal reflection fluorescence microscopy with a grating substrate,” Opt. Lett. 33, 255–257 (2008).
    [CrossRef]
  11. J. W. Goodman, Introduction to Fourier Optics (Roberts & Co., 2004).
  12. F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
    [CrossRef]
  13. J. K. Consulting, “Thin film design and applications,” http://www.kruschwitz.com/materials.htm.

2009 (1)

2008 (2)

M. G. Somekh, K. Hsu, and M. C. Pitter, “Resolution in structured illumination microscopy: a probabilistic approach,” J. Opt. Soc. Am. 25, 1319–1329 (2008).
[CrossRef]

A. Sentenac, K. Belkebir, H. Giovannini, and P. C. Chaumet, “Subdiffraction resolution in total internal reflection fluorescence microscopy with a grating substrate,” Opt. Lett. 33, 255–257 (2008).
[CrossRef]

2006 (1)

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

2000 (1)

M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc. 198, 82–87 (2000).
[CrossRef]

1999 (2)

R. Heintzmann and C. Cremer, “Laterally modulated excitation microscopy: Improvement of resolution by using a diffraction grating,” Proc. SPIE 3568, 185–196 (1999).
[CrossRef]

M. G. L. Gustafsson, D. A. Agard, and J. W. Sedat, “I5M: 3D widefield light microscopy with better than 100nm axial resolution,” J. Microsc. 195, 10–16 (1999).
[CrossRef]

1994 (2)

1993 (1)

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, “Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation,” Nature 366, 44–48 (1993).
[CrossRef]

Agard, D. A.

M. G. L. Gustafsson, D. A. Agard, and J. W. Sedat, “I5M: 3D widefield light microscopy with better than 100nm axial resolution,” J. Microsc. 195, 10–16 (1999).
[CrossRef]

Bailey, B.

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, “Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation,” Nature 366, 44–48 (1993).
[CrossRef]

Barnes, W. L.

Belkebir, K.

Chaumet, P. C.

Chuang, C. J.

Cremer, C.

R. Heintzmann and C. Cremer, “Laterally modulated excitation microscopy: Improvement of resolution by using a diffraction grating,” Proc. SPIE 3568, 185–196 (1999).
[CrossRef]

S. W. Hell, E. H. K. Stelzer, S. Lindek, and C. Cremer, “Confocal microscopy with an increased detection aperture: type-B 4Pi confocal microscopy,” Opt. Lett. 19, 222–224 (1994).
[CrossRef]

Espinos, J. P.

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

Farkas, D. L.

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, “Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation,” Nature 366, 44–48 (1993).
[CrossRef]

Giovannini, H.

Girardeau, T.

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

Gonzalez-Elipe, A. R.

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

Goodman, J. W.

J. W. Goodman, Introduction to Fourier Optics (Roberts & Co., 2004).

Gracia, F.

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

Gustafsson, M. G. L.

M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc. 198, 82–87 (2000).
[CrossRef]

M. G. L. Gustafsson, D. A. Agard, and J. W. Sedat, “I5M: 3D widefield light microscopy with better than 100nm axial resolution,” J. Microsc. 195, 10–16 (1999).
[CrossRef]

Heintzmann, R.

R. Heintzmann and C. Cremer, “Laterally modulated excitation microscopy: Improvement of resolution by using a diffraction grating,” Proc. SPIE 3568, 185–196 (1999).
[CrossRef]

Hell, S. W.

Holgado, J. P.

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

Hsu, K.

M. G. Somekh, K. Hsu, and M. C. Pitter, “Resolution in structured illumination microscopy: a probabilistic approach,” J. Opt. Soc. Am. 25, 1319–1329 (2008).
[CrossRef]

Lanni, F.

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, “Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation,” Nature 366, 44–48 (1993).
[CrossRef]

Lindek, S.

Liu, S.

Pitter, M. C.

M. G. Somekh, K. Hsu, and M. C. Pitter, “Resolution in structured illumination microscopy: a probabilistic approach,” J. Opt. Soc. Am. 25, 1319–1329 (2008).
[CrossRef]

Sedat, J. W.

M. G. L. Gustafsson, D. A. Agard, and J. W. Sedat, “I5M: 3D widefield light microscopy with better than 100nm axial resolution,” J. Microsc. 195, 10–16 (1999).
[CrossRef]

See, C. W.

Sentenac, A.

Somekh, M. G.

S. Liu, C. J. Chuang, C. W. See, G. Zoriniants, W. L. Barnes, and M. G. Somekh, “Double-grating-structured light microscope using plasmonic nanoparticle arrays,” Opt. Lett. 34, 1255–1257 (2009).
[CrossRef]

M. G. Somekh, K. Hsu, and M. C. Pitter, “Resolution in structured illumination microscopy: a probabilistic approach,” J. Opt. Soc. Am. 25, 1319–1329 (2008).
[CrossRef]

Stelzer, E. H. K.

Taylor, D. L.

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, “Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation,” Nature 366, 44–48 (1993).
[CrossRef]

Wichmann, Jan

Yubero, F.

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

Zoriniants, G.

J. Microsc. (2)

M. G. L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc. 198, 82–87 (2000).
[CrossRef]

M. G. L. Gustafsson, D. A. Agard, and J. W. Sedat, “I5M: 3D widefield light microscopy with better than 100nm axial resolution,” J. Microsc. 195, 10–16 (1999).
[CrossRef]

J. Opt. Soc. Am. (1)

M. G. Somekh, K. Hsu, and M. C. Pitter, “Resolution in structured illumination microscopy: a probabilistic approach,” J. Opt. Soc. Am. 25, 1319–1329 (2008).
[CrossRef]

Nature (1)

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, “Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation,” Nature 366, 44–48 (1993).
[CrossRef]

Opt. Lett. (4)

Proc. SPIE (1)

R. Heintzmann and C. Cremer, “Laterally modulated excitation microscopy: Improvement of resolution by using a diffraction grating,” Proc. SPIE 3568, 185–196 (1999).
[CrossRef]

Thin Solid Films (1)

F. Gracia, F. Yubero, J. P. Holgado, J. P. Espinos, A. R. Gonzalez-Elipe, and T. Girardeau, “SiO2/TiO2 thin films with variable refractive index prepared by ion beaminduced and plasma enhanced chemical vapour deposition,” Thin Solid Films 500, 19–26 (2006).
[CrossRef]

Other (3)

J. K. Consulting, “Thin film design and applications,” http://www.kruschwitz.com/materials.htm.

J. W. Goodman, Introduction to Fourier Optics (Roberts & Co., 2004).

The value of the k-vector is normalized with respect to 2π/λ so that it has a maximum value of 1 in air.

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