Abstract

We demonstrate the first use, to our knowledge, of a compact, diode-pumped, femtosecond fiber laser for third-harmonic generation (THG) microscopy. We discuss the utility of this technique, as well as the technical issues involved in using this compact source, and demonstrate the first use, to our knowledge, of imaging by THG backlighting.

© 1999 Optical Society of America

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References

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  1. J. A. Squier, M. Müller, G. J. Brakenhoff, K. R. Wilson, “Third harmonic generation microscopy,” Opt. Exp. 3, 315–324 (1998).
    [CrossRef]
  2. Y. Barad, H. Eisenberg, M. Horowitz, Y. Silberberg, “Non-linear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett. 70, 922–924 (1997).
    [CrossRef]
  3. M. Müller, J. A. Squier, K. R. Wilson, G. J. Brakenhoff, “3D microscopy of transparent objects using third harmonic generation,” J. Microsc. 191, 266–274 (1998).
    [CrossRef] [PubMed]
  4. J. A. Squier, M. Müller, “Third-harmonic generation imaging of laser-induced breakdown in glass,” Appl. Opt. 38, 5789–5794 (1999).
    [CrossRef]
  5. T. Y. F. Tsang, “Optical third harmonic generation at interfaces,” Phys. Rev. A 52, 4116–4125 (1995).
    [CrossRef] [PubMed]
  6. T. Y. F. Tsang, “Reflected optical harmonics from dielectric mirrors,” Appl. Opt. 33, 7720–7724 (1994).
    [CrossRef] [PubMed]
  7. D. Yelin, Y. Silberberg, Y. Barad, J. S. Patel, “Phase-matched third harmonic generation in a nematic liquid crystal,” Phys. Rev. Lett. 82, 3046–3049 (1999).
    [CrossRef]
  8. C. Preza, “Phase estimation using rotational diversity for differential interference contrast microscopy,” D. S. thesis (Department of Electrical Engineering, University of Washington, Saint Louis, Mo., 1998).
  9. M. E. Fermann, L.-M. Yang, M. L. Stock, M. T. Andrejco, “Environmentally stable Kerr-type mode-locked erbium fiber laser producing 360-fs pulses,” Opt. Lett. 19, 43–45 (1994).
    [CrossRef] [PubMed]
  10. R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
    [CrossRef]
  11. J. A. Curcio, C. C. Petty, “The near infrared absorption spectrum of liquid water,” J. Opt. Soc. Am. 41, 302–304 (1951).
    [CrossRef]
  12. A. Schönle, S. W. Hell, “Heating by absorption in the focus of an objective lens,” Opt. Lett. 23, 325–327 (1998).
    [CrossRef]
  13. D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15,741–15,746 (1998).
    [CrossRef]

1999

D. Yelin, Y. Silberberg, Y. Barad, J. S. Patel, “Phase-matched third harmonic generation in a nematic liquid crystal,” Phys. Rev. Lett. 82, 3046–3049 (1999).
[CrossRef]

J. A. Squier, M. Müller, “Third-harmonic generation imaging of laser-induced breakdown in glass,” Appl. Opt. 38, 5789–5794 (1999).
[CrossRef]

1998

A. Schönle, S. W. Hell, “Heating by absorption in the focus of an objective lens,” Opt. Lett. 23, 325–327 (1998).
[CrossRef]

D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15,741–15,746 (1998).
[CrossRef]

J. A. Squier, M. Müller, G. J. Brakenhoff, K. R. Wilson, “Third harmonic generation microscopy,” Opt. Exp. 3, 315–324 (1998).
[CrossRef]

M. Müller, J. A. Squier, K. R. Wilson, G. J. Brakenhoff, “3D microscopy of transparent objects using third harmonic generation,” J. Microsc. 191, 266–274 (1998).
[CrossRef] [PubMed]

1997

Y. Barad, H. Eisenberg, M. Horowitz, Y. Silberberg, “Non-linear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett. 70, 922–924 (1997).
[CrossRef]

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

1995

T. Y. F. Tsang, “Optical third harmonic generation at interfaces,” Phys. Rev. A 52, 4116–4125 (1995).
[CrossRef] [PubMed]

1994

1951

Andrejco, M. T.

Barad, Y.

D. Yelin, Y. Silberberg, Y. Barad, J. S. Patel, “Phase-matched third harmonic generation in a nematic liquid crystal,” Phys. Rev. Lett. 82, 3046–3049 (1999).
[CrossRef]

Y. Barad, H. Eisenberg, M. Horowitz, Y. Silberberg, “Non-linear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett. 70, 922–924 (1997).
[CrossRef]

Brakenhoff, G. J.

J. A. Squier, M. Müller, G. J. Brakenhoff, K. R. Wilson, “Third harmonic generation microscopy,” Opt. Exp. 3, 315–324 (1998).
[CrossRef]

M. Müller, J. A. Squier, K. R. Wilson, G. J. Brakenhoff, “3D microscopy of transparent objects using third harmonic generation,” J. Microsc. 191, 266–274 (1998).
[CrossRef] [PubMed]

Curcio, J. A.

DeLong, K. W.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Denk, W.

D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15,741–15,746 (1998).
[CrossRef]

Eisenberg, H.

Y. Barad, H. Eisenberg, M. Horowitz, Y. Silberberg, “Non-linear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett. 70, 922–924 (1997).
[CrossRef]

Fermann, M. E.

Fittinghoff, D. N.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Hell, S. W.

Helmchen, F.

D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15,741–15,746 (1998).
[CrossRef]

Horowitz, M.

Y. Barad, H. Eisenberg, M. Horowitz, Y. Silberberg, “Non-linear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett. 70, 922–924 (1997).
[CrossRef]

Kane, D. J.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Kleinfeld, D.

D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15,741–15,746 (1998).
[CrossRef]

Krumbügel, M. A.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Mitra, P. P.

D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15,741–15,746 (1998).
[CrossRef]

Müller, M.

J. A. Squier, M. Müller, “Third-harmonic generation imaging of laser-induced breakdown in glass,” Appl. Opt. 38, 5789–5794 (1999).
[CrossRef]

M. Müller, J. A. Squier, K. R. Wilson, G. J. Brakenhoff, “3D microscopy of transparent objects using third harmonic generation,” J. Microsc. 191, 266–274 (1998).
[CrossRef] [PubMed]

J. A. Squier, M. Müller, G. J. Brakenhoff, K. R. Wilson, “Third harmonic generation microscopy,” Opt. Exp. 3, 315–324 (1998).
[CrossRef]

Patel, J. S.

D. Yelin, Y. Silberberg, Y. Barad, J. S. Patel, “Phase-matched third harmonic generation in a nematic liquid crystal,” Phys. Rev. Lett. 82, 3046–3049 (1999).
[CrossRef]

Petty, C. C.

Preza, C.

C. Preza, “Phase estimation using rotational diversity for differential interference contrast microscopy,” D. S. thesis (Department of Electrical Engineering, University of Washington, Saint Louis, Mo., 1998).

Richman, B. A.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Schönle, A.

Silberberg, Y.

D. Yelin, Y. Silberberg, Y. Barad, J. S. Patel, “Phase-matched third harmonic generation in a nematic liquid crystal,” Phys. Rev. Lett. 82, 3046–3049 (1999).
[CrossRef]

Y. Barad, H. Eisenberg, M. Horowitz, Y. Silberberg, “Non-linear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett. 70, 922–924 (1997).
[CrossRef]

Squier, J. A.

J. A. Squier, M. Müller, “Third-harmonic generation imaging of laser-induced breakdown in glass,” Appl. Opt. 38, 5789–5794 (1999).
[CrossRef]

J. A. Squier, M. Müller, G. J. Brakenhoff, K. R. Wilson, “Third harmonic generation microscopy,” Opt. Exp. 3, 315–324 (1998).
[CrossRef]

M. Müller, J. A. Squier, K. R. Wilson, G. J. Brakenhoff, “3D microscopy of transparent objects using third harmonic generation,” J. Microsc. 191, 266–274 (1998).
[CrossRef] [PubMed]

Stock, M. L.

Sweetser, J. N.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Trebino, R.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Tsang, T. Y. F.

T. Y. F. Tsang, “Optical third harmonic generation at interfaces,” Phys. Rev. A 52, 4116–4125 (1995).
[CrossRef] [PubMed]

T. Y. F. Tsang, “Reflected optical harmonics from dielectric mirrors,” Appl. Opt. 33, 7720–7724 (1994).
[CrossRef] [PubMed]

Wilson, K. R.

M. Müller, J. A. Squier, K. R. Wilson, G. J. Brakenhoff, “3D microscopy of transparent objects using third harmonic generation,” J. Microsc. 191, 266–274 (1998).
[CrossRef] [PubMed]

J. A. Squier, M. Müller, G. J. Brakenhoff, K. R. Wilson, “Third harmonic generation microscopy,” Opt. Exp. 3, 315–324 (1998).
[CrossRef]

Yang, L.-M.

Yelin, D.

D. Yelin, Y. Silberberg, Y. Barad, J. S. Patel, “Phase-matched third harmonic generation in a nematic liquid crystal,” Phys. Rev. Lett. 82, 3046–3049 (1999).
[CrossRef]

Appl. Opt.

Appl. Phys. Lett.

Y. Barad, H. Eisenberg, M. Horowitz, Y. Silberberg, “Non-linear scanning laser microscopy by third harmonic generation,” Appl. Phys. Lett. 70, 922–924 (1997).
[CrossRef]

J. Microsc.

M. Müller, J. A. Squier, K. R. Wilson, G. J. Brakenhoff, “3D microscopy of transparent objects using third harmonic generation,” J. Microsc. 191, 266–274 (1998).
[CrossRef] [PubMed]

J. Opt. Soc. Am.

Opt. Exp.

J. A. Squier, M. Müller, G. J. Brakenhoff, K. R. Wilson, “Third harmonic generation microscopy,” Opt. Exp. 3, 315–324 (1998).
[CrossRef]

Opt. Lett.

Phys. Rev. A

T. Y. F. Tsang, “Optical third harmonic generation at interfaces,” Phys. Rev. A 52, 4116–4125 (1995).
[CrossRef] [PubMed]

Phys. Rev. Lett.

D. Yelin, Y. Silberberg, Y. Barad, J. S. Patel, “Phase-matched third harmonic generation in a nematic liquid crystal,” Phys. Rev. Lett. 82, 3046–3049 (1999).
[CrossRef]

Proc. Natl. Acad. Sci. USA

D. Kleinfeld, P. P. Mitra, F. Helmchen, W. Denk, “Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex,” Proc. Natl. Acad. Sci. USA 95, 15,741–15,746 (1998).
[CrossRef]

Rev. Sci. Instrum.

R. Trebino, K. W. DeLong, D. N. Fittinghoff, J. N. Sweetser, M. A. Krumbügel, B. A. Richman, D. J. Kane, “Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating,” Rev. Sci. Instrum. 68, 3277–3295 (1997).
[CrossRef]

Other

C. Preza, “Phase estimation using rotational diversity for differential interference contrast microscopy,” D. S. thesis (Department of Electrical Engineering, University of Washington, Saint Louis, Mo., 1998).

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Figures (9)

Fig. 1
Fig. 1

Contrast mechanism for THG. When an interface is to one side (a) or the other (c) of the focus, there is little THG, but when the interface is at the focus (b) there is substantial THG.

Fig. 2
Fig. 2

SHG FROG trace for fiber laser pulses. This is the raw trace, showing that the pulses have a strong ∼100-fs spike riding on a longer pedestal. The logarithm (base 10) of the intensity is plotted, with contours at steps of 0.5.

Fig. 3
Fig. 3

Fundamental spectrum of fiber laser. The peak intensity of the spectrum lies at 1562 nm. A modulated spectrum is typical for a second-order soliton, characteristic of the laser’s mode of operation.

Fig. 4
Fig. 4

Calculated third-harmonic spectrum. This spectrum is calculated by cubing the intensity of the fundamental at three times the wavelength, giving a peak intensity at 562 nm.

Fig. 5
Fig. 5

Experimental setup for THG imaging.

Fig. 6
Fig. 6

Emission spectrum of fiber laser in the 450–750-nm window. The spectrum exhibits multiple upconversion bands, as well as a weak signal from THG within the laser.

Fig. 7
Fig. 7

Leaf image section series. The chloroplasts of the leaf can be seen forming ringlike structures. (This is a false-color image, with the bright areas representing areas of third-harmonic emission.)

Fig. 8
Fig. 8

Third-harmonic backlighting. The cells are illuminated from behind by third-harmonic light generated from the surface of the coverslip.

Fig. 9
Fig. 9

Erythrocyte montage. These frames from a movie show the diffusion of red blood cells. (This is a false-color image, with the bright areas representing areas of third-harmonic emission.)

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