Abstract

We present a two-photon fluorescence microscope based on a three-port single-mode optical fiber coupler. It is found that the coupler behaves as a low-pass filter that can deliver an ultrashort-pulsed laser beam of as much as 150 mW of power in the wavelength range from 770 to 870 nm as well as collect a two-photon fluorescence signal in the visible range. As a result of using the fiber coupler, the new two-photon imaging system exhibts a number of advantages, including a compact arrangement, freedom from vibration from lasers and electronic devices, self-alignment, reduction of multiple scattering, and an enhanced optical sectioning effect. The effectiveness of the new instrument is demonstrated with a set of three-dimensional images of biological samples. This instrument may make two-photon fluorescence endoscopy possible for in vivo medical applications.

© 2002 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
    [CrossRef] [PubMed]
  2. T. Dabbs and M. Glass, Appl. Opt. 31, 705 (1992).
    [CrossRef] [PubMed]
  3. J. Benschop and G. van Rosmalen, Appl. Opt. 30, 1179 (1991).
    [CrossRef] [PubMed]
  4. S. Kimura and T. Wilson, Appl. Opt. 30, 2143 (1991).
    [CrossRef] [PubMed]
  5. M. Gu, C. J. R. Sheppard, and X. Gan, J. Opt. Soc. Am. A 8, 1755 (1991).
    [CrossRef]
  6. D. Bird and M. Gu, Appl. Opt. 41, 1852 (2002).
    [CrossRef] [PubMed]
  7. M. V. Iravani, Electron. Lett. 22, 103 (1986).
    [CrossRef]
  8. Y. Fujii and Y. Yamazaki, J. Microsc. (Oxford) 158, 145 (1989).
    [CrossRef]
  9. H. Zhou, M. Gu, and C. J. R. Sheppard, Optik 103, 45 (1996).
  10. M. Gu and D. K. Bird, Opt. Laser Technol. 30, 91 (1998).
    [CrossRef]
  11. K. König, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
    [CrossRef]
  12. M. Gu, Principles of Three-Dimensional Imaging in Confocal Midcroscopes (World Scientific, Singapore, 1996).
  13. G. P. Agrawal, Nonlinear Fiber Optics (Academic, San Diego, Calif., 1989).
  14. X. S. Gan and M. Gu, J. Appl. Phys. 87, 3214 (2000).
    [CrossRef]
  15. D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
    [CrossRef] [PubMed]
  16. G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
    [CrossRef] [PubMed]

2002 (1)

2000 (1)

X. S. Gan and M. Gu, J. Appl. Phys. 87, 3214 (2000).
[CrossRef]

1998 (1)

M. Gu and D. K. Bird, Opt. Laser Technol. 30, 91 (1998).
[CrossRef]

1997 (1)

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

1996 (1)

H. Zhou, M. Gu, and C. J. R. Sheppard, Optik 103, 45 (1996).

1995 (1)

K. König, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

1992 (1)

1991 (4)

J. Benschop and G. van Rosmalen, Appl. Opt. 30, 1179 (1991).
[CrossRef] [PubMed]

S. Kimura and T. Wilson, Appl. Opt. 30, 2143 (1991).
[CrossRef] [PubMed]

M. Gu, C. J. R. Sheppard, and X. Gan, J. Opt. Soc. Am. A 8, 1755 (1991).
[CrossRef]

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

1990 (1)

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

1989 (1)

Y. Fujii and Y. Yamazaki, J. Microsc. (Oxford) 158, 145 (1989).
[CrossRef]

1986 (1)

M. V. Iravani, Electron. Lett. 22, 103 (1986).
[CrossRef]

Agrawal, G. P.

G. P. Agrawal, Nonlinear Fiber Optics (Academic, San Diego, Calif., 1989).

Benschop, J.

Berns, M. W.

K. König, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Bird, D.

Bird, D. K.

M. Gu and D. K. Bird, Opt. Laser Technol. 30, 91 (1998).
[CrossRef]

Boppart, S. A.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Bouma, B. E.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Brezinski, M. E.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Chang, W.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Dabbs, T.

Denk, W.

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

Flotte, T.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Fujii, Y.

Y. Fujii and Y. Yamazaki, J. Microsc. (Oxford) 158, 145 (1989).
[CrossRef]

Fujimoto, J. G.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Gan, X.

Gan, X. S.

X. S. Gan and M. Gu, J. Appl. Phys. 87, 3214 (2000).
[CrossRef]

Glass, M.

Gregory, K.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Gu, M.

D. Bird and M. Gu, Appl. Opt. 41, 1852 (2002).
[CrossRef] [PubMed]

X. S. Gan and M. Gu, J. Appl. Phys. 87, 3214 (2000).
[CrossRef]

M. Gu and D. K. Bird, Opt. Laser Technol. 30, 91 (1998).
[CrossRef]

H. Zhou, M. Gu, and C. J. R. Sheppard, Optik 103, 45 (1996).

M. Gu, C. J. R. Sheppard, and X. Gan, J. Opt. Soc. Am. A 8, 1755 (1991).
[CrossRef]

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

Hee, M. R.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Huang, D.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Iravani, M. V.

M. V. Iravani, Electron. Lett. 22, 103 (1986).
[CrossRef]

Kimura, S.

König, K.

K. König, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Liang, H.

K. König, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Lin, C. P.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Pitris, C.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Puliafito, C. A.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Schuman, J. S.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Sheppard, C. J. R.

H. Zhou, M. Gu, and C. J. R. Sheppard, Optik 103, 45 (1996).

M. Gu, C. J. R. Sheppard, and X. Gan, J. Opt. Soc. Am. A 8, 1755 (1991).
[CrossRef]

Southern, J. F.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Stinson, W. G.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Strickler, J. H.

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

Swanson, E. A.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Tearney, G. J.

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Tromberg, B. J.

K. König, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

van Rosmalen, G.

Webb, W. W.

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

Wilson, T.

Yamazaki, Y.

Y. Fujii and Y. Yamazaki, J. Microsc. (Oxford) 158, 145 (1989).
[CrossRef]

Zhou, H.

H. Zhou, M. Gu, and C. J. R. Sheppard, Optik 103, 45 (1996).

Appl. Opt. (4)

Electron. Lett. (1)

M. V. Iravani, Electron. Lett. 22, 103 (1986).
[CrossRef]

J. Appl. Phys. (1)

X. S. Gan and M. Gu, J. Appl. Phys. 87, 3214 (2000).
[CrossRef]

J. Microsc. (Oxford) (1)

Y. Fujii and Y. Yamazaki, J. Microsc. (Oxford) 158, 145 (1989).
[CrossRef]

J. Opt. Soc. Am. A (1)

Nature (1)

K. König, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Opt. Laser Technol. (1)

M. Gu and D. K. Bird, Opt. Laser Technol. 30, 91 (1998).
[CrossRef]

Optik (1)

H. Zhou, M. Gu, and C. J. R. Sheppard, Optik 103, 45 (1996).

Science (3)

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

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

G. J. Tearney, M. E. Brezinski, B. E. Bouma, S. A. Boppart, C. Pitris, J. F. Southern, and J. G. Fujimoto, Science 276, 2037 (1997).
[CrossRef] [PubMed]

Other (2)

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

G. P. Agrawal, Nonlinear Fiber Optics (Academic, San Diego, Calif., 1989).

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (4)

Fig. 1
Fig. 1

Schematic diagram of the two-photon fluorescence microscope with a fiber coupler. PMT, photomultiplier tube. Other abbreviations defined in text.

Fig. 2
Fig. 2

Two-photon excitation efficiency of the two-photon fluorescence microscope with a fiber coupler. Inset (a) shows the axial responses to a thin fluorescent layer in the system when a coupler is used for (solid curve) an input power of 200 mW and (dotted curve) 1-m fiber delivery system employing a large-area detector without a pinhole. Inset (b) gives the dependence of the width of the axial response on the excitation power.

Fig. 3
Fig. 3

Images of 10µmdiameter fluorescent polymer microspheres in a two-photon fluorescence microscope with a fiber coupler. The size of the slices is 60 µm×60 µm, and the slice spacing is 1 µm. The excitation power is 8 mW at the focus.

Fig. 4
Fig. 4

Images of Griffithsia sea algae in a two-photon fluorescence microscope with a fiber coupler. The size of slices is 150 µm×150 µm, and the slice spacing is 1 µm. The excitation power is 8 mW at the focus.

Metrics