Abstract

We report on a novel and simple light source for short-wavelength two-photon excitation fluorescence microscopy based on the visible nonsolitonic radiation from a photonic crystal fiber. We demonstrate tunability of the light source by varying the wavelength and intensity of the Ti:Sapphire excitation light source. The visible nonsolitonic radiation is used as an excitation light source for two-photon fluorescence microscopy of tryptophan powder.

© 2005 Optical Society of America

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References

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  1. G. P. Agrawal, Nonlinear fiber optics, 3rd ed. (Academic Press, San Diego, 2001).
  2. J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
    [CrossRef] [PubMed]
  3. J. M. Dudley, L. Provino, N. Grossard, H. Maillotte, R. S. Windeler, B. J. Eggleton, and S. Coen, “Supercontinuum generation in air-silica microstructured fibers with nanosecond and femtosecond pulse pumping,” J. Opt. Soc. Am. B 19, 765–771 (2002);
    [CrossRef]
  4. A. V. Husakou and J. Herrmann, “Supercontinuum generation, four-wave mixing, and fission of higher-order solitons in photonic-crystal fibers,” J. Opt. Soc. Am. B 19, 2171–2182 (2002);
    [CrossRef]
  5. W. J. Wadsworth, A. Ortigosa-Blanch, J. C. Knight, T. A. Birks, T. P. M. Man, and P. S. Russell, “Supercontinuum generation in photonic crystal fibers and optical fiber tapers: a novel light source,” J. Opt. Soc. Am. B 19, 2148–2155 (2002);
    [CrossRef]
  6. J. K. Ranka, R. S. Windeler, and A. J. Stentz, “Optical properties of high-delta air-silica microstructure optical fibers,” Opt. Lett. 25, 796–798 (2000);
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  7. H. N. Paulsen, K. M. Hilligsoe, J. Thogersen, S. R. Keiding, and J. J. Larsen, “Coherent anti-Stokes Raman scattering microscopy with a photonic crystal fiber based light source,” Opt. Lett. 28, 1123–1125 (2003);
    [CrossRef] [PubMed]
  8. G. McConnell, “Confocal laser scanning fluorescence microscopy with a visible continuum source,” Opt. Express 12, 2844–2850 (2004);
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  9. C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
    [CrossRef]
  10. G. McConnell and E. Riis, “Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2,” Phys. Med. Biol. 49, 4757–4763 (2004);
    [CrossRef] [PubMed]
  11. K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
    [CrossRef]
  12. D. W. Piston, B. R. Masters, and W. W. Webb, “Three-dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two-photon excitation laser scanning microscopy,” J. Microsc. 178 (Pt 1), 20–27 (1995);
    [CrossRef]
  13. W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003);
    [CrossRef] [PubMed]
  14. A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. USA 99, 11014–11019 (2002);
    [CrossRef] [PubMed]
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    [CrossRef]
  17. K. M. Hilligsøe, H. N. Paulsen, J. Thøgersen, S. R. Keiding, and J. J. Larsen, “Initial steps of supercontinuum generation in photonic crystal fibers,” J. Opt. Soc. Am. B 20, 1887 (2003);
    [CrossRef]
  18. J. A. Gardecki and M. Maroncelli, “Set of secondary emission standards for calibration of the spectral responsivity in emission spectroscopy,” Appl. Spectrosc. 52, 1179–1189 (1998);
    [CrossRef]
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    [CrossRef]
  20. T. M. Fortier, J. Ye, S. T. Cundiff, and R. S. Windeler, “Nonlinear phase noise generated in air-silica microstructure fiber and its effect on carrier-envelope phase,” Opt. Lett. 27, 445–447 (2002);
    [CrossRef]
  21. A. Apolonski, B. Povazay, A. Unterhuber, W. Drexler, W. J. Wadsworth, J. C. Knight, and P. S. Russell, “Spectral shaping of supercontinuum in a cobweb photonic-crystal fiber with sub-20-fs pulses,” J. Opt. Soc. Am. B 19, 2165–2170 (2002);
    [CrossRef]
  22. G. Genty, M. Lehtonen, H. Ludvigsen, J. Broeng, and M. Kaivola, “Spectral broadening of femtosecond pulses into continuum radiation in microstructured fibers,” Opt. Express 10, 1083–1098 (2002);
    [PubMed]
  23. J. Bewersdorf and S. W. Hell, “Picosecond pulsed two-photon imaging with repetition rates of 200 and 400 MHz,” J. Microsc. 191, 28–38 (1998);
    [CrossRef]

2005 (1)

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
[CrossRef]

2004 (3)

G. McConnell, “Confocal laser scanning fluorescence microscopy with a visible continuum source,” Opt. Express 12, 2844–2850 (2004);
[CrossRef] [PubMed]

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

G. McConnell and E. Riis, “Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2,” Phys. Med. Biol. 49, 4757–4763 (2004);
[CrossRef] [PubMed]

2003 (3)

2002 (9)

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. USA 99, 11014–11019 (2002);
[CrossRef] [PubMed]

A. Ortigosa-Blanch, J. C. Knight, and P. S. J. Russell, “Pulse breaking and supercontinuum generation with 200-fs pump pulses in photonic crystal fibers,” J. Opt. Soc. Am. B 19, 2567–2572 (2002);
[CrossRef]

T. M. Fortier, J. Ye, S. T. Cundiff, and R. S. Windeler, “Nonlinear phase noise generated in air-silica microstructure fiber and its effect on carrier-envelope phase,” Opt. Lett. 27, 445–447 (2002);
[CrossRef]

A. Apolonski, B. Povazay, A. Unterhuber, W. Drexler, W. J. Wadsworth, J. C. Knight, and P. S. Russell, “Spectral shaping of supercontinuum in a cobweb photonic-crystal fiber with sub-20-fs pulses,” J. Opt. Soc. Am. B 19, 2165–2170 (2002);
[CrossRef]

G. Genty, M. Lehtonen, H. Ludvigsen, J. Broeng, and M. Kaivola, “Spectral broadening of femtosecond pulses into continuum radiation in microstructured fibers,” Opt. Express 10, 1083–1098 (2002);
[PubMed]

J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
[CrossRef] [PubMed]

J. M. Dudley, L. Provino, N. Grossard, H. Maillotte, R. S. Windeler, B. J. Eggleton, and S. Coen, “Supercontinuum generation in air-silica microstructured fibers with nanosecond and femtosecond pulse pumping,” J. Opt. Soc. Am. B 19, 765–771 (2002);
[CrossRef]

A. V. Husakou and J. Herrmann, “Supercontinuum generation, four-wave mixing, and fission of higher-order solitons in photonic-crystal fibers,” J. Opt. Soc. Am. B 19, 2171–2182 (2002);
[CrossRef]

W. J. Wadsworth, A. Ortigosa-Blanch, J. C. Knight, T. A. Birks, T. P. M. Man, and P. S. Russell, “Supercontinuum generation in photonic crystal fibers and optical fiber tapers: a novel light source,” J. Opt. Soc. Am. B 19, 2148–2155 (2002);
[CrossRef]

2000 (1)

1998 (2)

J. Bewersdorf and S. W. Hell, “Picosecond pulsed two-photon imaging with repetition rates of 200 and 400 MHz,” J. Microsc. 191, 28–38 (1998);
[CrossRef]

J. A. Gardecki and M. Maroncelli, “Set of secondary emission standards for calibration of the spectral responsivity in emission spectroscopy,” Appl. Spectrosc. 52, 1179–1189 (1998);
[CrossRef]

1995 (2)

J. N. Elgin, T. Brabec, and S. M. J. Kelly, “A Perturbative Theory of Soliton Propagation in the Presence of 3rd-Order Dispersion,” Opt. Commun. 114, 321–328 (1995);
[CrossRef]

D. W. Piston, B. R. Masters, and W. W. Webb, “Three-dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two-photon excitation laser scanning microscopy,” J. Microsc. 178 (Pt 1), 20–27 (1995);
[CrossRef]

Agrawal, G. P.

G. P. Agrawal, Nonlinear fiber optics, 3rd ed. (Academic Press, San Diego, 2001).

Apolonski, A.

Bewersdorf, J.

J. Bewersdorf and S. W. Hell, “Picosecond pulsed two-photon imaging with repetition rates of 200 and 400 MHz,” J. Microsc. 191, 28–38 (1998);
[CrossRef]

Birks, T. A.

Brabec, T.

J. N. Elgin, T. Brabec, and S. M. J. Kelly, “A Perturbative Theory of Soliton Propagation in the Presence of 3rd-Order Dispersion,” Opt. Commun. 114, 321–328 (1995);
[CrossRef]

Broeng, J.

Christie, R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003);
[CrossRef] [PubMed]

Coen, S.

Cundiff, S. T.

Davis, D. M.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Drexler, W.

Dudley, J. M.

Dunsby, C.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Eggleton, B. J.

Elgin, J. N.

J. N. Elgin, T. Brabec, and S. M. J. Kelly, “A Perturbative Theory of Soliton Propagation in the Presence of 3rd-Order Dispersion,” Opt. Commun. 114, 321–328 (1995);
[CrossRef]

Elson, D. S.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Fortier, T. M.

French, P. M. W.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Fukui, K.

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
[CrossRef]

Galletly, N.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Gardecki, J. A.

Genty, G.

Griebner, U.

J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
[CrossRef] [PubMed]

Grossard, N.

Hell, S. W.

J. Bewersdorf and S. W. Hell, “Picosecond pulsed two-photon imaging with repetition rates of 200 and 400 MHz,” J. Microsc. 191, 28–38 (1998);
[CrossRef]

Herrmann, J.

J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
[CrossRef] [PubMed]

A. V. Husakou and J. Herrmann, “Supercontinuum generation, four-wave mixing, and fission of higher-order solitons in photonic-crystal fibers,” J. Opt. Soc. Am. B 19, 2171–2182 (2002);
[CrossRef]

Higashi, T.

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
[CrossRef]

Hilligsoe, K. M.

Hilligsøe, K. M.

Husakou, A.

J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
[CrossRef] [PubMed]

Husakou, A. V.

Hyman, B. T.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003);
[CrossRef] [PubMed]

Isobe, K.

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
[CrossRef]

Itoh, K.

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
[CrossRef]

Kaivola, M.

Keiding, S. R.

Kelly, S. M. J.

J. N. Elgin, T. Brabec, and S. M. J. Kelly, “A Perturbative Theory of Soliton Propagation in the Presence of 3rd-Order Dispersion,” Opt. Commun. 114, 321–328 (1995);
[CrossRef]

Knight, J. C.

Korn, G.

J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
[CrossRef] [PubMed]

Lanigan, P. M. P.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Larsen, J. J.

Lehtonen, M.

Ludvigsen, H.

Maillotte, H.

Man, T. P. M.

Maroncelli, M.

Masters, B. R.

D. W. Piston, B. R. Masters, and W. W. Webb, “Three-dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two-photon excitation laser scanning microscopy,” J. Microsc. 178 (Pt 1), 20–27 (1995);
[CrossRef]

Matsunaga, S.

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
[CrossRef]

McCann, F.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

McConnell, G.

G. McConnell and E. Riis, “Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2,” Phys. Med. Biol. 49, 4757–4763 (2004);
[CrossRef] [PubMed]

G. McConnell, “Confocal laser scanning fluorescence microscopy with a visible continuum source,” Opt. Express 12, 2844–2850 (2004);
[CrossRef] [PubMed]

McGinty, J.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Munro, I.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Neil, M. A. A.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Nickel, D.

J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
[CrossRef] [PubMed]

Onfelt, B.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Ortigosa-Blanch, A.

Paulsen, H. N.

Piston, D. W.

D. W. Piston, B. R. Masters, and W. W. Webb, “Three-dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two-photon excitation laser scanning microscopy,” J. Microsc. 178 (Pt 1), 20–27 (1995);
[CrossRef]

Povazay, B.

Provino, L.

Ranka, J. K.

Requejo -Isidro, J.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Riis, E.

G. McConnell and E. Riis, “Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2,” Phys. Med. Biol. 49, 4757–4763 (2004);
[CrossRef] [PubMed]

Russell, P. S.

Russell, P. S. J.

Stentz, A. J.

Thogersen, J.

Thøgersen, J.

Treanor, B.

C. Dunsby, P. M. P. Lanigan, J. McGinty, D. S. Elson, J. Requejo -Isidro, I. Munro, N. Galletly, F. McCann, B. Treanor, B. Onfelt, D. M. Davis, M. A. A. Neil, and P. M. W. French, “An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy,” J. Phys. D 37, 3296 (2004);
[CrossRef]

Tromberg, B. J.

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. USA 99, 11014–11019 (2002);
[CrossRef] [PubMed]

Unterhuber, A.

Wadsworth, W. J.

Watanabe, W.

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2  44, L167–L169 (2005);
[CrossRef]

Webb, W. W.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003);
[CrossRef] [PubMed]

D. W. Piston, B. R. Masters, and W. W. Webb, “Three-dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two-photon excitation laser scanning microscopy,” J. Microsc. 178 (Pt 1), 20–27 (1995);
[CrossRef]

Williams, R. M.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003);
[CrossRef] [PubMed]

Windeler, R. S.

Ye, J.

Yeh, A.

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. USA 99, 11014–11019 (2002);
[CrossRef] [PubMed]

Yu Nikitin, A.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003);
[CrossRef] [PubMed]

Zhavoronkov, N.

J. Herrmann, U. Griebner, N. Zhavoronkov, A. Husakou, D. Nickel, J. C. Knight, W. J. Wadsworth, P. S. Russell, and G. Korn, “Experimental evidence for supercontinuum generation by fission of higher-order solitons in photonic fibers,” Phys. Rev. Lett. 88, 173901 (2002);
[CrossRef] [PubMed]

Zipfel, W. R.

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Zoumi, A.

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W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003);
[CrossRef] [PubMed]

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. USA 99, 11014–11019 (2002);
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Other (2)

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Figures (6)

Fig. 1.
Fig. 1.

SEM image of the PCF (NL-1.5-670, Crystal Fibre A/S) “holey” region (left) and the core (right) [15].

Fig. 2.
Fig. 2.

Left: Output spectrum of the PCF (length=13 cm) with increasing laser input power (excitation wavelength=720 nm). The white dashed line denotes the zero-dispersion wavelength (ZDW) of the PCF. Right: Output spectrum of the PCF with varying laser input wavelength (excitation average power=300 mW).

Fig. 3.
Fig. 3.

Two-photon excitation fluorescence microscope setup coupled with the PCF. The inset shows the visible non-solitonic radiation generated along the 13-cm long PCF when excited with near infrared laser.

Fig. 4.
Fig. 4.

Left: Experimentally measured (circles) and standard (solid line) fluorescence spectra of tryptophan powder [18]. Right: Double-logarithmic plot of the tryptophan powder fluorescence intensity versus excitation intensity. The slope of the best -fit line is 2.16.

Fig. 5.
Fig. 5.

XY scan (left) and XZ scan (right) two-photon excitation fluorescence images of tryptophan powder using a Fluor 40X/1.30NA oil immersion objective lens.

Fig. 6.
Fig. 6.

Upper: Time trace of the relative intensity fluctuations of the PCF output. Lower: Power spectrum obtained by Fourier transforming the time trace (50,000 points).

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