Abstract

We present measurements of two-photon excitation (TPE) spectra of various fluorescent proteins with nonlinear Fourier-transform spectroscopy. By using an ultrabroadband laser pulse with a spectrum ranging from 700 to 1100nm, the absolute TPE spectra of six typical fluorescent proteins (SeBFP, Sapphire, eGFP, eCFP, Venus, DsRed) were measured with high spectral resolution.

© 2010 Optical Society of America

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  1. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
    [CrossRef] [PubMed]
  2. C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
    [CrossRef] [PubMed]
  3. 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. 44, L167–L169 (2005).
    [CrossRef]
  4. P. Allcock and D. L. Andrews, “Two-photon fluorescence: resonance energy transfer,” J. Chem. Phys. 108, 3089–3095 (1998).
    [CrossRef]
  5. G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
    [CrossRef] [PubMed]
  6. G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
    [CrossRef]
  7. W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21, 1369–1377 (2003).
    [CrossRef] [PubMed]
  8. E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
    [CrossRef] [PubMed]
  9. M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
    [CrossRef] [PubMed]
  10. H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
    [CrossRef] [PubMed]
  11. M. Bellini, A. Bartoli, and T. W. Hänsch, “Two-photon Fourier spectroscopy with femtosecond light pulses,” Opt. Lett. 22, 540–542 (1997).
    [CrossRef] [PubMed]
  12. J. P. Ogilvie, K. J. Kubarych, A. Alexandrou, and M. Joffre, “Fourier transform measurement of two-photon excitation spectra: applications to microscopy and optimal control,” Opt. Lett. 30, 911–913 (2005).
    [CrossRef] [PubMed]
  13. K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
    [CrossRef] [PubMed]
  14. K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
    [CrossRef]
  15. T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
    [CrossRef]
  16. J. K. Ranka, A. L. Gaeta, A. Baltuska, M. S. Pshenichnikov, and D. A. Wiersma, “Autocorrelation measurement of 6fs pulses based on the two-photon-induced photocurrent in a GaAsP photodiode,” Opt. Lett. 22, 1344–1366 (1997).
    [CrossRef]
  17. A. Baltuska, M. S. Pshenichnikov, and D. A. Wiersma, “Amplitude and phase characterization of 4.5fs pulses by frequency-resolved optical gating,” Opt. Lett. 23, 1474–1476 (1998).
    [CrossRef]
  18. N. S. Makarov, M. Drobizhev, and A. Rebane, “Two-photon absorption standards in the 550–1600nm excitation wavelength range,” Opt. Express 16, 4029–4047 (2008).
    [CrossRef] [PubMed]
  19. See, for example, http://www.clonetech.com
  20. S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
    [CrossRef] [PubMed]
  21. R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
    [CrossRef] [PubMed]

2009

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
[CrossRef] [PubMed]

2008

H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

N. S. Makarov, M. Drobizhev, and A. Rebane, “Two-photon absorption standards in the 550–1600nm excitation wavelength range,” Opt. Express 16, 4029–4047 (2008).
[CrossRef] [PubMed]

2005

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

J. P. Ogilvie, K. J. Kubarych, A. Alexandrou, and M. Joffre, “Fourier transform measurement of two-photon excitation spectra: applications to microscopy and optimal control,” Opt. Lett. 30, 911–913 (2005).
[CrossRef] [PubMed]

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. 44, L167–L169 (2005).
[CrossRef]

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

2003

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21, 1369–1377 (2003).
[CrossRef] [PubMed]

2002

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
[CrossRef] [PubMed]

2001

G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
[CrossRef]

1999

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

1998

1997

1996

C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
[CrossRef] [PubMed]

1990

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

Acker, H.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Alexandrou, A.

Allcock, P.

P. Allcock and D. L. Andrews, “Two-photon fluorescence: resonance energy transfer,” J. Chem. Phys. 108, 3089–3095 (1998).
[CrossRef]

Ando, R.

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
[CrossRef] [PubMed]

Andrews, D. L.

P. Allcock and D. L. Andrews, “Two-photon fluorescence: resonance energy transfer,” J. Chem. Phys. 108, 3089–3095 (1998).
[CrossRef]

Baltuska, A.

Bartoli, A.

Bellini, M.

Berchner-Pfannschmidt, U.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Bestvater, F.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Blab, G. A.

G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
[CrossRef]

Bonsma, S.

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

Cognet, L.

G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
[CrossRef]

Denk, W.

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

Drobizhev, M.

M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
[CrossRef] [PubMed]

N. S. Makarov, M. Drobizhev, and A. Rebane, “Two-photon absorption standards in the 550–1600nm excitation wavelength range,” Opt. Express 16, 4029–4047 (2008).
[CrossRef] [PubMed]

Ellisman, M. H.

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

Fan, G. Y.

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

Feurer, T.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Fujisaki, H.

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

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. 44, L167–L169 (2005).
[CrossRef]

Gaeta, A. L.

Gallus, J.

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

Hacker, M.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Hama, H.

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
[CrossRef] [PubMed]

Hänsch, T. W.

Harms, G. S.

G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
[CrossRef]

Heckel-Pompey, A.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

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. 44, L167–L169 (2005).
[CrossRef]

Hosoi, H.

H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
[CrossRef] [PubMed]

Hughes, T. E.

M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
[CrossRef] [PubMed]

Ibata, K.

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

Isobe, K.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

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. 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. 44, L167–L169 (2005).
[CrossRef]

Jezowski, S.

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

Joffre, M.

Kannari, F.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

Kawano, H.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

Könz, F.

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

Kubarych, K. J.

Kubota, M.

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

Lommerse, P. H. M.

G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
[CrossRef]

Makarov, N. S.

M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
[CrossRef] [PubMed]

N. S. Makarov, M. Drobizhev, and A. Rebane, “Two-photon absorption standards in the 550–1600nm excitation wavelength range,” Opt. Express 16, 4029–4047 (2008).
[CrossRef] [PubMed]

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. 44, L167–L169 (2005).
[CrossRef]

Midorikawa, K.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

Mikoshiba, K.

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

Miyawaki, A.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
[CrossRef] [PubMed]

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
[CrossRef] [PubMed]

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

Mizuno, H.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
[CrossRef] [PubMed]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
[CrossRef] [PubMed]

Nagai, T.

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

Ogilvie, J. P.

Porwol, T.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Pshenichnikov, M. S.

Purchase, R.

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

Ranka, J. K.

Rebane, A.

M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
[CrossRef] [PubMed]

N. S. Makarov, M. Drobizhev, and A. Rebane, “Two-photon absorption standards in the 550–1600nm excitation wavelength range,” Opt. Express 16, 4029–4047 (2008).
[CrossRef] [PubMed]

Schmidt, T.

G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
[CrossRef]

Shear, J. B.

C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
[CrossRef] [PubMed]

Spiess, E.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Stobrawa, G.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[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]

Suda, A.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

Sun Park, E.

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

Tahara, T.

H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
[CrossRef] [PubMed]

Tanaka, M.

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses,” Opt. Express 17, 13737–13746 (2009).
[CrossRef] [PubMed]

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

Tillo, S.

M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
[CrossRef] [PubMed]

Toth, K.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Tsay, R. K.

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

Tsien, R. Y.

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

Völker, S.

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

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. 44, L167–L169 (2005).
[CrossRef]

Webb, W. W.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21, 1369–1377 (2003).
[CrossRef] [PubMed]

C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
[CrossRef] [PubMed]

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

Wiersma, D. A.

Williams, R. M.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21, 1369–1377 (2003).
[CrossRef] [PubMed]

C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
[CrossRef] [PubMed]

Wotzlaw, C.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

Xu, C.

C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
[CrossRef] [PubMed]

Yamaguchi, S.

H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
[CrossRef] [PubMed]

Yamamoto-Hino, M.

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
[CrossRef] [PubMed]

Zipfel, W.

C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
[CrossRef] [PubMed]

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21, 1369–1377 (2003).
[CrossRef] [PubMed]

Biophys. J.

G. Y. Fan, H. Fujisaki, A. Miyawaki, R. K. Tsay, R. Y. Tsien, and M. H. Ellisman, “Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with Cameleons,” Biophys. J. 76, 2412–2420 (1999).
[CrossRef] [PubMed]

Chem. Phys. Chem.

S. Bonsma, R. Purchase, S. Jezowski, J. Gallus, F. Könz, and S. Völker, “Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning,” Chem. Phys. Chem. 6, 838–849 (2005).
[CrossRef] [PubMed]

Chem. Phys. Lett.

G. A. Blab, P. H. M. Lommerse, L. Cognet, G. S. Harms, and T. Schmidt, “Two-photon excitation action cross-sections of the autofluorescent proteins,” Chem. Phys. Lett. 350, 71–77 (2001).
[CrossRef]

J. Chem. Phys.

P. Allcock and D. L. Andrews, “Two-photon fluorescence: resonance energy transfer,” J. Chem. Phys. 108, 3089–3095 (1998).
[CrossRef]

J. Microsc.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217, 200–204 (2005).
[CrossRef] [PubMed]

J. Phys. Chem. B

M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes, and A. Rebane, “Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins,” J. Phys. Chem. B 113, 855–859 (2009).
[CrossRef] [PubMed]

H. Hosoi, S. Yamaguchi, H. Mizuno, A. Miyawaki, and T. Tahara, “Hidden electronic excited state of enhanced green fluorescent protein,” J. Phys. Chem. B 112, 2761–2763 (2008).
[CrossRef] [PubMed]

Jpn. J. Appl. Phys.

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. 44, L167–L169 (2005).
[CrossRef]

Nat. Biotechnol.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21, 1369–1377 (2003).
[CrossRef] [PubMed]

T. Nagai, K. Ibata, E. Sun Park, M. Kubota, K. Mikoshiba, and A. Miyawaki, “A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications,” Nat. Biotechnol. 20, 87–90 (2002).
[CrossRef]

Opt. Express

Opt. Lett.

Phys. Rev. A

K. Isobe, A. Suda, M. Tanaka, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, and K. Midorikawa, “Fourier-transform spectroscopy combined with a 5fs broadband pulse for multispectral nonlinear microscopy,” Phys. Rev. A 77, 063832(2008).
[CrossRef]

Proc. Natl. Acad. Sci. USA

C. Xu, W. Zipfel, J. B. Shear, R. M. Williams, and W. W. Webb, “Multiphoton fluorescence excitation: New spectral windows for biological nonlinear microscopy,” Proc. Natl. Acad. Sci. USA 93, 10763–10768 (1996).
[CrossRef] [PubMed]

R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno, and A. Miyawaki, “An optical marker based in the UV-induced green-to-red photoconversion of a fluorescent protein,” Proc. Natl. Acad. Sci. USA 99, 12651–12656 (2002).
[CrossRef] [PubMed]

Science

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

Other

See, for example, http://www.clonetech.com

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Figures (7)

Fig. 1
Fig. 1

Second-order IAC traces for (a) TPEF and (b) reference SH signal, and Fourier spectra of (c) TPEF and (d) reference SH signal.

Fig. 2
Fig. 2

Experimental setup. BS, beam splitter; DF, dichroic filter; OB, objective lens; PMT, photomultiplier tube; BPF, bandpass filter; SH, second harmonic; SLM, spatial light modulator.

Fig. 3
Fig. 3

(a) Comparison of measured SH spectrum (solid curve) with calculated SH spectrum (dotted curve) from fundamental spectrum of broadband laser pulse shown in (b).

Fig. 4
Fig. 4

Fourier spectra of (a) reference SH signal and (b) TPEF from Venus for various values of GDD applied to a broadband excitation pulse and (c) resultant TPE spectra of Venus.

Fig. 5
Fig. 5

Absolute TPE spectra of fluorescent proteins SeBFP, Sapphire, eCFP, eGFP, Venus, and DsRed.

Fig. 6
Fig. 6

Comparison of TPE cross section of eGFP measured with other techniques. (a) Ref. [6], (b) Ref. [7], (c) Ref. [10], and (d) this work. It should be noted that panel (c) shows two-photon absorption cross section of eGFP.

Fig. 7
Fig. 7

Comparison of TPE spectra (dotted curves) with OPE spectra (solid curves) of (a) eGFP and (b) Venus.

Equations (9)

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S ( 2 ) ( τ ) = | P ( t , τ ) | 2 d t ,
P ( t , τ ) = δ ( t t 1 ) E ( t 1 , τ ) 2 d t 1 ,
E ( t , τ ) = [ A ( t ) + A ( t τ ) exp ( i ω 0 τ ) ] exp ( i ω 0 t ) .
P ( TPEF ) ( t , τ ) = R ( TPEF ) ( t t 1 ) E ( t 1 , τ ) 2 d t 1 .
S ( 2 ) ( τ ) = d t [ | H 1 ( t ) | 2 + 4 | H 2 ( t , τ ) | 2 + | H 3 ( t , τ ) | 2 + 2 { H 1 ( t ) H 2 * ( t , τ ) + H 2 ( t ) H 3 * ( t , τ ) } exp ( i ω 0 τ ) + H 1 ( t ) H 3 * ( t , τ ) exp ( i 2 ω 0 τ ) + c . c . ] ,
H 1 ( t ) = R ( TPEF ) ( t t 1 ) A ( t 1 ) 2 exp ( i 2 ω 0 t 1 ) d t 1 ,
H 2 ( t , τ ) = R ( TPEF ) ( t t 1 ) A ( t 1 ) A ( t 1 τ ) exp ( i 2 ω 0 t 1 ) d t 1 ,
H 3 ( t , τ ) = R ( TPEF ) ( t t 1 ) A ( t 1 τ ) 2 exp ( i 2 ω 0 t 1 ) d t 1 .
S ˜ 2 ω 0 ( 2 ) ( Ω ) = H 1 ( t ) H 3 * ( t , τ ) exp ( i 2 ω 0 τ ) d t exp ( i Ω τ ) d τ = | R ˜ ( TPEF ) ( Ω ) | 2 | A ˜ ( 2 ) ( Ω 2 ω 0 ) | 2 ,

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