Abstract

It is shown that one can obtain absolute two-photon fluorescence excitation (TPE) cross sections without prior knowledge of the temporal profile of the excitation pulse. It is sufficient to record the fluorescence generated by two spatially superimposed but temporally shifted halves of the excitation beam as a function of their relative delay. Measurements employed a Michelson interferometer with a mode-locked Ti:sapphire laser as the excitation source. The TPE cross section for fluorescein [(46 ± 10) × 10−58 m4 s/photon] obtained at 782 nm with this technique agrees with single-mode cw excitation data. We find no indication of a deviation from the power-squared dependence of TPE.

© 1995 Optical Society of America

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  1. W. Denk, J. H. Strickler, W. W. Webb, Science 248, 73 (1990).
    [CrossRef] [PubMed]
  2. W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
    [CrossRef] [PubMed]
  3. W. Denk, D. W. Piston, W. W. Webb, in The Handbook of Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), p. 445.
  4. R. Yuste, W. Denk, Nature (London) 375, 682 (1995).
    [CrossRef] [PubMed]
  5. D. W. Piston, B. R. Masters, W. W. Webb, J. Microsc. 178, 20 (1995).
    [CrossRef] [PubMed]
  6. R. M. Williams, D. W. Piston, W. W. Webb, FASEB J. 8, 804 (1994).
    [PubMed]
  7. A. J. Twarowski, D. S. Kliger, Chem. Phys. 20, 259 (1977)M. Sheik-Bahae, A. A. Said, T. Wei, D. Hagan, E. W. Van Stryland, IEEE J. Quantum Electron. 26, 760 (1990).
    [CrossRef]
  8. J. P. Hermann, J. Ducuing, Phys. Rev. A 5, 2557 (1972).
    [CrossRef]
  9. R. Loudon, The Quantum Theory of Light (Clarendon, Oxford, 1983), p. 82.
  10. S. M. Kennedy, F. E. Lytle, Anal. Chem. 58, 2643 (1986)R. D. Jones, P. R. Callis, J. Appl. Phys. 64, 4301 (1988).
    [CrossRef]
  11. M. Muller, J. Squier, G. J. Brakenhoff, Opt. Lett. 20, 1038 (1995).
    [CrossRef] [PubMed]
  12. J. Diels, in Dye Laser Principles, F. J. Duarte, L. W. Hillman, eds. (Academic, San Diego, Calif., 1990), p. 104.
  13. W. W. Webb, C. Xu, “Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 nm to 990 nm,” J. Opt. Soc. Am. B (to be published)C. Xu, J. Guild, W. W. Webb, Biophys. J. 68, A197 (1995).

1995 (3)

R. Yuste, W. Denk, Nature (London) 375, 682 (1995).
[CrossRef] [PubMed]

D. W. Piston, B. R. Masters, W. W. Webb, J. Microsc. 178, 20 (1995).
[CrossRef] [PubMed]

M. Muller, J. Squier, G. J. Brakenhoff, Opt. Lett. 20, 1038 (1995).
[CrossRef] [PubMed]

1994 (2)

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

R. M. Williams, D. W. Piston, W. W. Webb, FASEB J. 8, 804 (1994).
[PubMed]

1990 (1)

W. Denk, J. H. Strickler, W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

1986 (1)

S. M. Kennedy, F. E. Lytle, Anal. Chem. 58, 2643 (1986)R. D. Jones, P. R. Callis, J. Appl. Phys. 64, 4301 (1988).
[CrossRef]

1977 (1)

A. J. Twarowski, D. S. Kliger, Chem. Phys. 20, 259 (1977)M. Sheik-Bahae, A. A. Said, T. Wei, D. Hagan, E. W. Van Stryland, IEEE J. Quantum Electron. 26, 760 (1990).
[CrossRef]

1972 (1)

J. P. Hermann, J. Ducuing, Phys. Rev. A 5, 2557 (1972).
[CrossRef]

Brakenhoff, G. J.

Delaney, K. R.

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

Denk, W.

R. Yuste, W. Denk, Nature (London) 375, 682 (1995).
[CrossRef] [PubMed]

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

W. Denk, J. H. Strickler, W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

W. Denk, D. W. Piston, W. W. Webb, in The Handbook of Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), p. 445.

Diels, J.

J. Diels, in Dye Laser Principles, F. J. Duarte, L. W. Hillman, eds. (Academic, San Diego, Calif., 1990), p. 104.

Ducuing, J.

J. P. Hermann, J. Ducuing, Phys. Rev. A 5, 2557 (1972).
[CrossRef]

Gelperin, A.

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

Hermann, J. P.

J. P. Hermann, J. Ducuing, Phys. Rev. A 5, 2557 (1972).
[CrossRef]

Kennedy, S. M.

S. M. Kennedy, F. E. Lytle, Anal. Chem. 58, 2643 (1986)R. D. Jones, P. R. Callis, J. Appl. Phys. 64, 4301 (1988).
[CrossRef]

Kleinfeld, D.

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

Kliger, D. S.

A. J. Twarowski, D. S. Kliger, Chem. Phys. 20, 259 (1977)M. Sheik-Bahae, A. A. Said, T. Wei, D. Hagan, E. W. Van Stryland, IEEE J. Quantum Electron. 26, 760 (1990).
[CrossRef]

Loudon, R.

R. Loudon, The Quantum Theory of Light (Clarendon, Oxford, 1983), p. 82.

Lytle, F. E.

S. M. Kennedy, F. E. Lytle, Anal. Chem. 58, 2643 (1986)R. D. Jones, P. R. Callis, J. Appl. Phys. 64, 4301 (1988).
[CrossRef]

Masters, B. R.

D. W. Piston, B. R. Masters, W. W. Webb, J. Microsc. 178, 20 (1995).
[CrossRef] [PubMed]

Muller, M.

Piston, D. W.

D. W. Piston, B. R. Masters, W. W. Webb, J. Microsc. 178, 20 (1995).
[CrossRef] [PubMed]

R. M. Williams, D. W. Piston, W. W. Webb, FASEB J. 8, 804 (1994).
[PubMed]

W. Denk, D. W. Piston, W. W. Webb, in The Handbook of Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), p. 445.

Squier, J.

Strickler, J. H.

W. Denk, J. H. Strickler, W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

Strowbridge, B. W.

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

Tank, D. W.

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

Twarowski, A. J.

A. J. Twarowski, D. S. Kliger, Chem. Phys. 20, 259 (1977)M. Sheik-Bahae, A. A. Said, T. Wei, D. Hagan, E. W. Van Stryland, IEEE J. Quantum Electron. 26, 760 (1990).
[CrossRef]

Webb, W. W.

D. W. Piston, B. R. Masters, W. W. Webb, J. Microsc. 178, 20 (1995).
[CrossRef] [PubMed]

R. M. Williams, D. W. Piston, W. W. Webb, FASEB J. 8, 804 (1994).
[PubMed]

W. Denk, J. H. Strickler, W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

W. Denk, D. W. Piston, W. W. Webb, in The Handbook of Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), p. 445.

W. W. Webb, C. Xu, “Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 nm to 990 nm,” J. Opt. Soc. Am. B (to be published)C. Xu, J. Guild, W. W. Webb, Biophys. J. 68, A197 (1995).

Williams, R. M.

R. M. Williams, D. W. Piston, W. W. Webb, FASEB J. 8, 804 (1994).
[PubMed]

Xu, C.

W. W. Webb, C. Xu, “Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 nm to 990 nm,” J. Opt. Soc. Am. B (to be published)C. Xu, J. Guild, W. W. Webb, Biophys. J. 68, A197 (1995).

Yuste, R.

R. Yuste, W. Denk, Nature (London) 375, 682 (1995).
[CrossRef] [PubMed]

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

Anal. Chem. (1)

S. M. Kennedy, F. E. Lytle, Anal. Chem. 58, 2643 (1986)R. D. Jones, P. R. Callis, J. Appl. Phys. 64, 4301 (1988).
[CrossRef]

Chem. Phys. (1)

A. J. Twarowski, D. S. Kliger, Chem. Phys. 20, 259 (1977)M. Sheik-Bahae, A. A. Said, T. Wei, D. Hagan, E. W. Van Stryland, IEEE J. Quantum Electron. 26, 760 (1990).
[CrossRef]

FASEB J. (1)

R. M. Williams, D. W. Piston, W. W. Webb, FASEB J. 8, 804 (1994).
[PubMed]

J. Microsc. (1)

D. W. Piston, B. R. Masters, W. W. Webb, J. Microsc. 178, 20 (1995).
[CrossRef] [PubMed]

J. Neurosci. Methods (1)

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste, J. Neurosci. Methods 54, 151 (1994).
[CrossRef] [PubMed]

Nature (1)

R. Yuste, W. Denk, Nature (London) 375, 682 (1995).
[CrossRef] [PubMed]

Opt. Lett. (1)

Phys. Rev. A (1)

J. P. Hermann, J. Ducuing, Phys. Rev. A 5, 2557 (1972).
[CrossRef]

Science (1)

W. Denk, J. H. Strickler, W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

Other (4)

W. Denk, D. W. Piston, W. W. Webb, in The Handbook of Confocal Microscopy, J. Pawley, ed. (Plenum, New York, 1995), p. 445.

R. Loudon, The Quantum Theory of Light (Clarendon, Oxford, 1983), p. 82.

J. Diels, in Dye Laser Principles, F. J. Duarte, L. W. Hillman, eds. (Academic, San Diego, Calif., 1990), p. 104.

W. W. Webb, C. Xu, “Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 nm to 990 nm,” J. Opt. Soc. Am. B (to be published)C. Xu, J. Guild, W. W. Webb, Biophys. J. 68, A197 (1995).

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