Abstract

The imaging performances of multiphoton excitation and confocal laser scanning microscopy are herby considered: in typical experimental imaging conditions, a small finite amount of photon reaches the detector giving shot-noise fluctuations which affects the signal acquired. A significant detriment in the high frequencies transmission capability is obtained. In order to partially recover the high frequencies information lost, the insertion of a pupil plane filter in the microscope illumination light pathway on the objective lens is proposed. We demonstrate high-frequency and resolution enhancement in the case of linear and non linear fluorescence microscope approach under shot-noise condition.

© 2009 Optical Society of America

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    [CrossRef] [PubMed]
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2008 (2)

P. P. Mondal and A. Diaspro, "Lateral resolution improvement in two photon excitation microscopy by aperture engineering," Opt. Commun. 281,1855-1859 (2008).
[CrossRef]

P. P. Mondal, G. Vicidomini, and A. Diaspro, "Image reconstruction for multiphoton fluorescence microscopy," Appl. Phys. Lett. 92, 103902 (2008).
[CrossRef]

2007 (2)

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[CrossRef] [PubMed]

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

2006 (3)

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, "Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy," Biophys. J. 91, 4258-4272 (2006).
[CrossRef] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, "Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)," Nat. Methods 3, 793-796 (2006).
[CrossRef] [PubMed]

O. Haeberl and B. Simon, "Improving lateral resolution in confocal fluorescence microscopy using laterally interfering excitation beams," Opt. Commun. 259, 400-408 (2006).
[CrossRef]

2005 (3)

Y. Garini, B. J. Vermolen and I. T. Young, "From micro to nano: recent advances in high-resolution microscopy," Curr. Opin. Biotechnol. 16, 3-12 (2005).
[CrossRef] [PubMed]

A. Diaspro, G. Chirico, and M. Collini, "Two photon fluorescence excitation and related techniques in biological microscopy," Q. Rev. Biophys. 38, 97-166 (2005).
[CrossRef]

C. Ibáñez-López, G. Saavedra, G. Boyer, and M. Martinez-Corral, "Quasi-isotropic 3-D resolution in two-photon scanning microscopy," Opt. Express 13, 6168-6174 (2005).
[CrossRef] [PubMed]

2004 (2)

2002 (1)

M. R. Arnison and C. J. R. Sheppard, "A 3d vectorial optical transfer function suitable for arbitrary pupil functions," Opt. Comm. 211, 53-63 (2002).
[CrossRef]

2001 (2)

2000 (2)

1999 (2)

H. J. Koester, D. Baur, R. Uhl, and S. W. Hell, "Ca2+ Fluorescence Imaging with Pico- and Femtosecond Two-Photon Excitation: Signal and Photodamage," Biophys. J. 77, 2226-2236 (1999).
[CrossRef] [PubMed]

M. G. L. Gustafsson, "Extended resolution fluorescence microscopy," Curr. Opin. Struct. Biol. 9,627-634 (1999).
[CrossRef] [PubMed]

1998 (1)

E. H. K. Steltzer, "Contrast, resolution, pixilation, dynamic range and signal-to-noise ratio: fundamental limits to resolution in fluorescence light microscopy," J. Microsc. 189, 15-24 (1998).
[CrossRef]

1997 (1)

1996 (1)

I. T. Young, "Quantitative Microscopy," IEEE Eng. Med. Biol. 15, 59-66 (1996).
[CrossRef]

1995 (1)

D. R. Sandison, D. W. Piston, R. M. Williams, and W. W. Webb, "Quantitative comparison of background rejection, signal-to-noise ratio, and resolution in confocal and full-field laser scanning microscopes," Appl. Opt. 34, (1995).
[CrossRef] [PubMed]

1993 (1)

M. Gu and C. J. R. Sheppard, "Effects of a finite-sized pinhole on 3D image formation in confocal two-photon fluorescence microscopy," J. Mod. Opt. 40, 2009-2024 (1993).
[CrossRef]

1992 (1)

M. Gu and C. J. R. Sheppard, "Three-dimensional optical transfer function in a fiber-optical confocal fluorescence microscope using annular lenses," J. Opt. Soc. Am. A 9, 1993-1999 (1992).
[CrossRef]

1991 (1)

K. Carlsson, "The influence of specimen refractive index, detector signal integration and non uniform scan spees on the imaging properties in confocal microscopy," J. Microsc. 163, 167-178 (1991).
[CrossRef]

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, "Two-Photon Laser Scanning Fluorescence Microscopy," Science 248, 73-76 (1990).
[CrossRef] [PubMed]

1986 (1)

I. J. Cox and C. J. R. Sheppard, "Information capacity and resolution in an optical system," J. Opt. Soc. Am. 3, (1986).
[CrossRef]

1984 (1)

K. Lange and R. Carson, "EM Reconstruction Algorithms for emission and transmission tomography," J. Comput. Assist. Tomogr. 8, 306-316 (1984).
[PubMed]

1982 (1)

1978 (1)

C. J. R. Sheppard and T. Wilson, "Image formation in scanning microscopes with partially coherent source and detector," Optica Acta 25, 315-325 (1978).
[CrossRef]

1964 (1)

1959 (2)

E. Wolf, "Electromagnetic diffraction in optical systems I. An integral representation of the image field" Proc. R. Soc. London Ser. A 253, 349-357 (1959).
[CrossRef]

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. R. Soc. London Ser. A 253, 358-379 (1959).
[CrossRef]

1956 (1)

1952 (1)

G. Toraldo di Francia, "Nuovo pupille superrisolvente," Atti Fond.Giorgio Ronchi 7, 366-372 (1952).

Andresen, M.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Arnison, M. R.

M. R. Arnison and C. J. R. Sheppard, "A 3d vectorial optical transfer function suitable for arbitrary pupil functions," Opt. Comm. 211, 53-63 (2002).
[CrossRef]

Bates, M.

M. J. Rust, M. Bates, and X. Zhuang, "Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)," Nat. Methods 3, 793-796 (2006).
[CrossRef] [PubMed]

Baur, D.

H. J. Koester, D. Baur, R. Uhl, and S. W. Hell, "Ca2+ Fluorescence Imaging with Pico- and Femtosecond Two-Photon Excitation: Signal and Photodamage," Biophys. J. 77, 2226-2236 (1999).
[CrossRef] [PubMed]

Bock, H.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Boyer, G.

Caorsi, V.

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[CrossRef] [PubMed]

Carlsson, K.

K. Carlsson, "The influence of specimen refractive index, detector signal integration and non uniform scan spees on the imaging properties in confocal microscopy," J. Microsc. 163, 167-178 (1991).
[CrossRef]

Carson, R.

K. Lange and R. Carson, "EM Reconstruction Algorithms for emission and transmission tomography," J. Comput. Assist. Tomogr. 8, 306-316 (1984).
[PubMed]

Chirico, G.

A. Diaspro, G. Chirico, and M. Collini, "Two photon fluorescence excitation and related techniques in biological microscopy," Q. Rev. Biophys. 38, 97-166 (2005).
[CrossRef]

Choudhury, A.

Collini, M.

A. Diaspro, G. Chirico, and M. Collini, "Two photon fluorescence excitation and related techniques in biological microscopy," Q. Rev. Biophys. 38, 97-166 (2005).
[CrossRef]

Cox, I. J.

I. J. Cox and C. J. R. Sheppard, "Information capacity and resolution in an optical system," J. Opt. Soc. Am. 3, (1986).
[CrossRef]

I. J. Cox, C. J. R. Sheppard, and T. Wilson, "Reappraisal of arrays of concentric annuli as superresolving filters," J. Opt. Soc. Am. 72, 1287-1291(1982).
[CrossRef]

Day, R. N.

G. Patterson, R. N. Day, and D. Piston, "Fluorescent protein spectra," J.Cell Science 114, 837-838 (2001).
[PubMed]

Denk, W.

W. Denk, J. H. Strickler, and W. W. Webb, "Two-Photon Laser Scanning Fluorescence Microscopy," Science 248, 73-76 (1990).
[CrossRef] [PubMed]

Diaspro, A.

P. P. Mondal, G. Vicidomini, and A. Diaspro, "Image reconstruction for multiphoton fluorescence microscopy," Appl. Phys. Lett. 92, 103902 (2008).
[CrossRef]

P. P. Mondal and A. Diaspro, "Lateral resolution improvement in two photon excitation microscopy by aperture engineering," Opt. Commun. 281,1855-1859 (2008).
[CrossRef]

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[CrossRef] [PubMed]

A. Diaspro, G. Chirico, and M. Collini, "Two photon fluorescence excitation and related techniques in biological microscopy," Q. Rev. Biophys. 38, 97-166 (2005).
[CrossRef]

Eggeling, C.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Egner, A.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Garini, Y.

Y. Garini, B. J. Vermolen and I. T. Young, "From micro to nano: recent advances in high-resolution microscopy," Curr. Opin. Biotechnol. 16, 3-12 (2005).
[CrossRef] [PubMed]

Geisler, C.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Girirajan, T. P. K.

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, "Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy," Biophys. J. 91, 4258-4272 (2006).
[CrossRef] [PubMed]

Gu, M.

M. Gu and C. J. R. Sheppard, "Effects of a finite-sized pinhole on 3D image formation in confocal two-photon fluorescence microscopy," J. Mod. Opt. 40, 2009-2024 (1993).
[CrossRef]

M. Gu and C. J. R. Sheppard, "Three-dimensional optical transfer function in a fiber-optical confocal fluorescence microscope using annular lenses," J. Opt. Soc. Am. A 9, 1993-1999 (1992).
[CrossRef]

Gustafsson, M. G. L.

M. G. L. Gustafsson, "Extended resolution fluorescence microscopy," Curr. Opin. Struct. Biol. 9,627-634 (1999).
[CrossRef] [PubMed]

Haeberl, O.

O. Haeberl and B. Simon, "Improving lateral resolution in confocal fluorescence microscopy using laterally interfering excitation beams," Opt. Commun. 259, 400-408 (2006).
[CrossRef]

Hell, S. W.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

M. Nagorni and S. W. Hell, "Coherent use of opposing lenses for axial resolution increase in fluorescence microscopy. I. Comparative study of concepts," J. Opt. Soc. Am. A 18, 36-48 (2001).
[CrossRef]

H. J. Koester, D. Baur, R. Uhl, and S. W. Hell, "Ca2+ Fluorescence Imaging with Pico- and Femtosecond Two-Photon Excitation: Signal and Photodamage," Biophys. J. 77, 2226-2236 (1999).
[CrossRef] [PubMed]

Hess, S. T.

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, "Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy," Biophys. J. 91, 4258-4272 (2006).
[CrossRef] [PubMed]

Ibáñez-López, C.

Jakobs, S.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Jovin, T. M.

Juskaitis, R.

Koester, H. J.

H. J. Koester, D. Baur, R. Uhl, and S. W. Hell, "Ca2+ Fluorescence Imaging with Pico- and Femtosecond Two-Photon Excitation: Signal and Photodamage," Biophys. J. 77, 2226-2236 (1999).
[CrossRef] [PubMed]

Krol, S.

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[CrossRef] [PubMed]

Laczik, Z. J.

Lange, K.

K. Lange and R. Carson, "EM Reconstruction Algorithms for emission and transmission tomography," J. Comput. Assist. Tomogr. 8, 306-316 (1984).
[PubMed]

Martinez-Corral, M.

Mason, M. D.

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, "Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy," Biophys. J. 91, 4258-4272 (2006).
[CrossRef] [PubMed]

McConnell, G.

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[CrossRef] [PubMed]

McCutchen, C. W.

Medda, R.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Mondal, P. P.

P. P. Mondal, G. Vicidomini, and A. Diaspro, "Image reconstruction for multiphoton fluorescence microscopy," Appl. Phys. Lett. 92, 103902 (2008).
[CrossRef]

P. P. Mondal and A. Diaspro, "Lateral resolution improvement in two photon excitation microscopy by aperture engineering," Opt. Commun. 281,1855-1859 (2008).
[CrossRef]

Nagorni, M.

Neil, M.

O’Neill, E. L.

Patterson, G.

G. Patterson, R. N. Day, and D. Piston, "Fluorescent protein spectra," J.Cell Science 114, 837-838 (2001).
[PubMed]

Patterson, G. H.

G. H. Patterson and D. W. Piston, "Photobleaching in Two-Photon Excitation Microscopy," Biophys J. 78, 2159-2162 (2000).
[CrossRef] [PubMed]

Piston, D.

G. Patterson, R. N. Day, and D. Piston, "Fluorescent protein spectra," J.Cell Science 114, 837-838 (2001).
[PubMed]

Piston, D. W.

G. H. Patterson and D. W. Piston, "Photobleaching in Two-Photon Excitation Microscopy," Biophys J. 78, 2159-2162 (2000).
[CrossRef] [PubMed]

D. R. Sandison, D. W. Piston, R. M. Williams, and W. W. Webb, "Quantitative comparison of background rejection, signal-to-noise ratio, and resolution in confocal and full-field laser scanning microscopes," Appl. Opt. 34, (1995).
[CrossRef] [PubMed]

Richards, B.

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. R. Soc. London Ser. A 253, 358-379 (1959).
[CrossRef]

Ronzitti, E.

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[CrossRef] [PubMed]

Rust, M. J.

M. J. Rust, M. Bates, and X. Zhuang, "Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)," Nat. Methods 3, 793-796 (2006).
[CrossRef] [PubMed]

Saavedra, G.

Sandison, D. R.

D. R. Sandison, D. W. Piston, R. M. Williams, and W. W. Webb, "Quantitative comparison of background rejection, signal-to-noise ratio, and resolution in confocal and full-field laser scanning microscopes," Appl. Opt. 34, (1995).
[CrossRef] [PubMed]

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Schönle, A.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Sheppard, C. J. R.

C. J. R. Sheppard and A. Choudhury, "Annular pupils, radial polarization, and superresolution," Appl. Opt. 43, 4322-4327 (2004).
[CrossRef] [PubMed]

M. R. Arnison and C. J. R. Sheppard, "A 3d vectorial optical transfer function suitable for arbitrary pupil functions," Opt. Comm. 211, 53-63 (2002).
[CrossRef]

M. Gu and C. J. R. Sheppard, "Effects of a finite-sized pinhole on 3D image formation in confocal two-photon fluorescence microscopy," J. Mod. Opt. 40, 2009-2024 (1993).
[CrossRef]

M. Gu and C. J. R. Sheppard, "Three-dimensional optical transfer function in a fiber-optical confocal fluorescence microscope using annular lenses," J. Opt. Soc. Am. A 9, 1993-1999 (1992).
[CrossRef]

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[CrossRef]

I. J. Cox, C. J. R. Sheppard, and T. Wilson, "Reappraisal of arrays of concentric annuli as superresolving filters," J. Opt. Soc. Am. 72, 1287-1291(1982).
[CrossRef]

C. J. R. Sheppard and T. Wilson, "Image formation in scanning microscopes with partially coherent source and detector," Optica Acta 25, 315-325 (1978).
[CrossRef]

Simon, B.

O. Haeberl and B. Simon, "Improving lateral resolution in confocal fluorescence microscopy using laterally interfering excitation beams," Opt. Commun. 259, 400-408 (2006).
[CrossRef]

Steltzer, E. H. K.

E. H. K. Steltzer, "Contrast, resolution, pixilation, dynamic range and signal-to-noise ratio: fundamental limits to resolution in fluorescence light microscopy," J. Microsc. 189, 15-24 (1998).
[CrossRef]

Stiel, A.-C.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

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]

Toraldo di Francia, G.

G. Toraldo di Francia, "Nuovo pupille superrisolvente," Atti Fond.Giorgio Ronchi 7, 366-372 (1952).

Uhl, R.

H. J. Koester, D. Baur, R. Uhl, and S. W. Hell, "Ca2+ Fluorescence Imaging with Pico- and Femtosecond Two-Photon Excitation: Signal and Photodamage," Biophys. J. 77, 2226-2236 (1999).
[CrossRef] [PubMed]

Vermolen, B. J.

Y. Garini, B. J. Vermolen and I. T. Young, "From micro to nano: recent advances in high-resolution microscopy," Curr. Opin. Biotechnol. 16, 3-12 (2005).
[CrossRef] [PubMed]

Verveer, P. J.

Vicidomini, G.

P. P. Mondal, G. Vicidomini, and A. Diaspro, "Image reconstruction for multiphoton fluorescence microscopy," Appl. Phys. Lett. 92, 103902 (2008).
[CrossRef]

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[CrossRef] [PubMed]

von Middendorff, C.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Webb, W. W.

D. R. Sandison, D. W. Piston, R. M. Williams, and W. W. Webb, "Quantitative comparison of background rejection, signal-to-noise ratio, and resolution in confocal and full-field laser scanning microscopes," Appl. Opt. 34, (1995).
[CrossRef] [PubMed]

W. Denk, J. H. Strickler, and W. W. Webb, "Two-Photon Laser Scanning Fluorescence Microscopy," Science 248, 73-76 (1990).
[CrossRef] [PubMed]

Wenzel, D.

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

Williams, R. M.

D. R. Sandison, D. W. Piston, R. M. Williams, and W. W. Webb, "Quantitative comparison of background rejection, signal-to-noise ratio, and resolution in confocal and full-field laser scanning microscopes," Appl. Opt. 34, (1995).
[CrossRef] [PubMed]

Wilson, T.

Wolf, E.

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. R. Soc. London Ser. A 253, 358-379 (1959).
[CrossRef]

E. Wolf, "Electromagnetic diffraction in optical systems I. An integral representation of the image field" Proc. R. Soc. London Ser. A 253, 349-357 (1959).
[CrossRef]

Young, I. T.

Y. Garini, B. J. Vermolen and I. T. Young, "From micro to nano: recent advances in high-resolution microscopy," Curr. Opin. Biotechnol. 16, 3-12 (2005).
[CrossRef] [PubMed]

I. T. Young, "Quantitative Microscopy," IEEE Eng. Med. Biol. 15, 59-66 (1996).
[CrossRef]

Zhuang, X.

M. J. Rust, M. Bates, and X. Zhuang, "Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)," Nat. Methods 3, 793-796 (2006).
[CrossRef] [PubMed]

Appl. Opt. (2)

D. R. Sandison, D. W. Piston, R. M. Williams, and W. W. Webb, "Quantitative comparison of background rejection, signal-to-noise ratio, and resolution in confocal and full-field laser scanning microscopes," Appl. Opt. 34, (1995).
[CrossRef] [PubMed]

C. J. R. Sheppard and A. Choudhury, "Annular pupils, radial polarization, and superresolution," Appl. Opt. 43, 4322-4327 (2004).
[CrossRef] [PubMed]

Appl. Phys. Lett. (1)

P. P. Mondal, G. Vicidomini, and A. Diaspro, "Image reconstruction for multiphoton fluorescence microscopy," Appl. Phys. Lett. 92, 103902 (2008).
[CrossRef]

Biophys J. (1)

G. H. Patterson and D. W. Piston, "Photobleaching in Two-Photon Excitation Microscopy," Biophys J. 78, 2159-2162 (2000).
[CrossRef] [PubMed]

Biophys. J. (3)

H. J. Koester, D. Baur, R. Uhl, and S. W. Hell, "Ca2+ Fluorescence Imaging with Pico- and Femtosecond Two-Photon Excitation: Signal and Photodamage," Biophys. J. 77, 2226-2236 (1999).
[CrossRef] [PubMed]

A. Egner, C. Geisler, C. von Middendorff, H. Bock, D. Wenzel, R. Medda, M. Andresen, A.-C. Stiel, S. Jakobs, C. Eggeling, A. Schönle, and S. W. Hell, "Fluorescence nanoscopy in whole cells by asynchronous localization of photoswitching emitters," Biophys. J. 93, 3285-3290 (2007).
[CrossRef] [PubMed]

S. T. Hess, T. P. K. Girirajan, and M. D. Mason, "Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy," Biophys. J. 91, 4258-4272 (2006).
[CrossRef] [PubMed]

Curr. Opin. Biotechnol. (1)

Y. Garini, B. J. Vermolen and I. T. Young, "From micro to nano: recent advances in high-resolution microscopy," Curr. Opin. Biotechnol. 16, 3-12 (2005).
[CrossRef] [PubMed]

Curr. Opin. Struct. Biol. (1)

M. G. L. Gustafsson, "Extended resolution fluorescence microscopy," Curr. Opin. Struct. Biol. 9,627-634 (1999).
[CrossRef] [PubMed]

Giorgio Ronchi (1)

G. Toraldo di Francia, "Nuovo pupille superrisolvente," Atti Fond.Giorgio Ronchi 7, 366-372 (1952).

IEEE Eng. Med. Biol. (1)

I. T. Young, "Quantitative Microscopy," IEEE Eng. Med. Biol. 15, 59-66 (1996).
[CrossRef]

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K. Lange and R. Carson, "EM Reconstruction Algorithms for emission and transmission tomography," J. Comput. Assist. Tomogr. 8, 306-316 (1984).
[PubMed]

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E. H. K. Steltzer, "Contrast, resolution, pixilation, dynamic range and signal-to-noise ratio: fundamental limits to resolution in fluorescence light microscopy," J. Microsc. 189, 15-24 (1998).
[CrossRef]

K. Carlsson, "The influence of specimen refractive index, detector signal integration and non uniform scan spees on the imaging properties in confocal microscopy," J. Microsc. 163, 167-178 (1991).
[CrossRef]

J. Mod. Opt. (1)

M. Gu and C. J. R. Sheppard, "Effects of a finite-sized pinhole on 3D image formation in confocal two-photon fluorescence microscopy," J. Mod. Opt. 40, 2009-2024 (1993).
[CrossRef]

J. Opt. Soc. Am. (4)

J. Opt. Soc. Am. A (3)

J.Cell Science (1)

G. Patterson, R. N. Day, and D. Piston, "Fluorescent protein spectra," J.Cell Science 114, 837-838 (2001).
[PubMed]

Microsc. Res. Tech. (1)

V. Caorsi, E. Ronzitti, G. Vicidomini, S. Krol, G. McConnell, and A. Diaspro "FRET Measurements on Fuzzy Fluorescent Nanostructures," Microsc. Res. Tech. 70, 452-458 (2007).
[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, 793-796 (2006).
[CrossRef] [PubMed]

Opt. Comm. (1)

M. R. Arnison and C. J. R. Sheppard, "A 3d vectorial optical transfer function suitable for arbitrary pupil functions," Opt. Comm. 211, 53-63 (2002).
[CrossRef]

Opt. Commun. (2)

O. Haeberl and B. Simon, "Improving lateral resolution in confocal fluorescence microscopy using laterally interfering excitation beams," Opt. Commun. 259, 400-408 (2006).
[CrossRef]

P. P. Mondal and A. Diaspro, "Lateral resolution improvement in two photon excitation microscopy by aperture engineering," Opt. Commun. 281,1855-1859 (2008).
[CrossRef]

Opt. Express (2)

Opt. Lett. (1)

Optica Acta (1)

C. J. R. Sheppard and T. Wilson, "Image formation in scanning microscopes with partially coherent source and detector," Optica Acta 25, 315-325 (1978).
[CrossRef]

Proc. R. Soc. London Ser. A (2)

E. Wolf, "Electromagnetic diffraction in optical systems I. An integral representation of the image field" Proc. R. Soc. London Ser. A 253, 349-357 (1959).
[CrossRef]

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. R. Soc. London Ser. A 253, 358-379 (1959).
[CrossRef]

Q. Rev. Biophys. (1)

A. Diaspro, G. Chirico, and M. Collini, "Two photon fluorescence excitation and related techniques in biological microscopy," Q. Rev. Biophys. 38, 97-166 (2005).
[CrossRef]

Science (1)

W. Denk, J. H. Strickler, and W. W. Webb, "Two-Photon Laser Scanning Fluorescence Microscopy," Science 248, 73-76 (1990).
[CrossRef] [PubMed]

Other (9)

T. Wilson and C. J. R. Sheppard, Theory and practice of scanning optical microscopy (AcademicPress, London,1984).

J. E. N. Jonkman and E. H. K. Steltzer, "Resolution and contrast in confocal Two-Photon Microscopy" in Confocal and Two Photons: Foundations, Applications and Advances, A.Diaspro Ed. (Wiley-Liss, 2002).

J. B. Pawley, "Fundamental limits in confocal microscopy" in Handbook of biological confocal microscopy, J. B. Pawley Ed (Springer, 2006) Chap.2.

F. Cella, E. Ronzitti, G. Vicidomini, P. P. Mondal, and A. Diaspro "Studying the illumination puzzle towards an isotropic increase of optical resolution," Proc. of SPIE 6861 (2008).
[CrossRef]

M. Gu, Principles of Three-Dimensional Imaging in Confocal Microscopes (World Scientific, 1996).

C. J. R. Sheppard, X. Gan, M. Gu, and M. Roy, "Signal-to-Noise Ratio in Confocal Microscopes" in Handbook of biological confocal microscopy, J. B. Pawley Ed (Springer, 2006) Chap.22.
[CrossRef]

A. Diaspro, G. Chirico, C. Usai, P. Ramoino, and J. Dobrucki, "Photobleaching," in Handbook of biological confocal microscopy, J. B. Pawley Ed (Springer, 2006) Chap.16.
[CrossRef]

P. J. Shaw, "Comparison of Widefield/Deconvolution and confocal microscopy for three-dimensional Imaging," in Handbook of biological confocal microscopy, J. B. Pawley Ed (Springer, 2006) Chap.23.
[CrossRef]

E. H. K. Steltzer, "The intermediate optical system of laser-scanning confocal microscopes", in Handbook of biological confocal microscopy," J. B. Pawley Ed (Springer, 2006), Chap. 9.

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Figures (9)

Fig. 1.
Fig. 1.

Annular filter scheme

Fig. 2.
Fig. 2.

Theoretical PSF for 2PE (a) and CLSM (c) configuration; PSF associated with a finite expected number of photons np =20photons (b),(d). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm (scale bar equal to 100nm)

Fig. 3.
Fig. 3.

Theoretical OTF for 2PE (a) and CLSM (b) configuration; PSF associated with a finite expected number of photons np =20photons (c),(d). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm (bar line is 1.2μm-1).

Fig 4.
Fig 4.

Axial OTF in 2PE (a) and in CLSM (b) scheme for different photon influxes to the detector assuming a particular noise realization; OTF high-frequency region in the insets (c)(d). Dots line in the inset indicate the cut-off frequencies estimated (black dots theoretical, red dots np =20photons, blue dots np =100photons). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm.

Fig. 5.
Fig. 5.

Radial OTF in 2PE (a) and in CLSM (b) scheme for different photon influxes to the detector assuming a particular noise realization; OTF high-frequency region in the insets (c)(d). Dots line in the inset indicate the cut-off frequencies estimated (black dots theoretical, red dots np =20photons, blue dots np =100photons). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm.

Fig. 6.
Fig. 6.

Axial OTF for 2PE (a) and CLSM (b) scheme for different filters.. OTF high-frequency region in the insets (c)(d). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm.

Fig. 7.
Fig. 7.

Radial OTF for 2PE (a) and CLSM (b) scheme for different filters. OTF high-frequency region in the insets (c)(d). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm.

Fig. 8.
Fig. 8.

Axial OTF in 2PE (a) and in CLSM (b) scheme for filter configuration in shot-noise condition assuming a particular noise realization; OTF high-frequency region in the insets (c)(d).Dots line in the inset indicate the cut-off frequencies estimated (black theoretical, red np =20photons, blue filter). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm.

Fig. 9.
Fig. 9.

Radial OTF in 2PE (a) and in CLSM (b) scheme for filter configuration in shot-noise condition assuming a particular noise realization; OTF high-frequency region in the insets (c)(d). Dots line in the inset indicate the cut-off frequencies estimated (black theoretical, red np =20photons, blue filter). NA=1.4, n=1.518, excitation wavelength 900nm TPE, 400nm CLSM; detection wavelength 500nm.

Tables (3)

Tables Icon

Table 1. Estimation of the effective cut-off frequency values varying the number of photons emitted for pixel (NA1.4; refractive index 1,51; excitation wavelength 900nm 2PE, 400nm CLSM; detection wavelength 500nm)..

Tables Icon

Table 2. Effective axial cut-off frequencies assuming a finite amount of photons to the detector in filtering scheme. Data in round brackets represent the filter percentage improvement respect to the unfiltered case.

Tables Icon

Table 3. Effective radial cut-off frequencies assuming a finite amount of photons to the detector in filtering scheme. Data in round brackets represent the filter percentage improvement respect to the unfiltered case.

Equations (9)

Equations on this page are rendered with MathJax. Learn more.

PS F system ( u , v , ϕ P ) = [ h ex ( u , v , ϕ P ) 2 ] m · [ D ( v ) h em ( u β , v β , ϕ P ) 2 ]
{ u = k r P cos ϑ P sin 2 α v = k r P sin θ P sin α
PS F CLSM ( u , v , ϕ P ) = ( h ex ( u , v , ϕ P ) 2 ) · ( h ex ( u β , v β , ϕ P ) 2 )
PS F 2 PE ( u , v , ϕ P ) = ( h ex ( u , v , ϕ P ) 2 ) 2
h ( u , v ) 2 = 1 2 π 0 2 π E ( u , v , ϕ P ) 2 d ϕ P
{ e x = iA π 0 α 0 2 π K ( θ , ϕ ) cos θ sin θ { cos θ + ( 1 cos θ ) sin 2 ϕ } e ik r p cos ε dθdϕ e y = iA π 0 α 0 2 π K ( θ , ϕ ) cos θ sin θ ( 1 cos θ ) cos ϕ sin ϕ e ik r p cos ε dθdϕ e z = iA π 0 α 0 2 π K ( θ , ϕ ) cos θ sin 2 θ cos ϕ e ik r p cos ε dθdϕ
cos ε = cos θ cos θ P + sin θ sin θ P cos ( ϕ ϕ P )
K ( θ ) = { 0 α 2 ( 1 C 100 ) < θ < α 2 ( 1 + C 100 ) 1 otherwise
P ( n ) = ( γt ) n e γt n !

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