Abstract

A fast handheld two-photon fiber-optic fluorescence endoscope for three-dimensional (3D) in vivo cellular imaging is developed. The compact handheld probe of the two-photon endoscope can simply be placed into contact with the target tissue to reveal clear 3D surface and subsurface histological images without biopsy. The new system has, to the best of our knowledge, the largest field of view (FOV) of 475μm×475μm and a 3D imaging volume larger than 475μm×475μm×250μm. A real-time two-photon fluorescence image is displayed at 0.4mm2s. The lateral and axial resolutions of the two-photon fluorescence endoscope are better than 1 and 14.5μm, respectively.

© 2008 Optical Society of America

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  1. W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
    [CrossRef] [PubMed]
  2. B. A. Flusberg, E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, Nat. Methods 12, 941 (2005).
    [CrossRef]
  3. L. Fu and M. Gu, J. Microsc. 226, 195 (2007).
    [CrossRef] [PubMed]
  4. F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
    [CrossRef] [PubMed]
  5. D. Bird and M. Gu, Opt. Lett. 27, 1031 (2002).
    [CrossRef]
  6. D. Bird and M. Gu, Opt. Lett. 28, 1552 (2003).
    [CrossRef] [PubMed]
  7. L. Fu, X. Gan, and M. Gu, Opt. Lett. 30, 385 (2005).
    [CrossRef] [PubMed]
  8. L. Fu, X. Gan, and M. Gu, Opt. Express 13, 5528 (2005).
    [CrossRef] [PubMed]
  9. L. Fu, A. Jain, C. Cranfield, H. Xie, and M. Gu, J. Biomed. Opt. 12, 040501 (2007).
    [CrossRef] [PubMed]
  10. L. Fu and M. Gu, Opt. Lett. 31, 1471 (2006).
    [CrossRef] [PubMed]
  11. W. Piyawattanametha, R. P. J. Barretto, T. H. Ko, B. A. Flusberg, E. D. Cocker, H. Ra, D. Lee, and O. Solgaard, Opt. Lett. 31, 2018 (2006).
    [CrossRef] [PubMed]
  12. B. A. Flusberg, J. C. Jung, E. D. Cocker, E. P. Anderson, and M. J. Schnitzer, Opt. Lett. 30, 2272 (2005).
    [CrossRef] [PubMed]
  13. M. T. Myaing, D. J. MacDonald, and X. Li, Opt. Lett. 31, 1076 (2006).
    [CrossRef] [PubMed]
  14. S. W. Clark, F. O. Ilday, and F. W. Wise, Opt. Lett. 26, 1320 (2001).
    [CrossRef]
  15. M. Lelek, E. Suran, F. Louradour, A. Barthelemy, B. Viellerobe, and F. Lacombe, Opt. Express 15, 10154 (2007).
    [CrossRef] [PubMed]
  16. M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
    [CrossRef] [PubMed]
  17. M. Gu, Advanced Optical Imaging Theory, Springer Series in Optical Sciences 75 (Springer-Verlag, 2000).

2007 (4)

L. Fu and M. Gu, J. Microsc. 226, 195 (2007).
[CrossRef] [PubMed]

L. Fu, A. Jain, C. Cranfield, H. Xie, and M. Gu, J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

M. Lelek, E. Suran, F. Louradour, A. Barthelemy, B. Viellerobe, and F. Lacombe, Opt. Express 15, 10154 (2007).
[CrossRef] [PubMed]

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

2006 (3)

2005 (4)

2003 (1)

2002 (1)

2001 (2)

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

S. W. Clark, F. O. Ilday, and F. W. Wise, Opt. Lett. 26, 1320 (2001).
[CrossRef]

1990 (1)

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

Anderson, E. P.

Anlauf, M.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Barretto, R. P. J.

Bartenstein, P.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Barthelemy, A.

Bird, D.

Cheung, E. L. M.

B. A. Flusberg, E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, Nat. Methods 12, 941 (2005).
[CrossRef]

Clark, S. W.

Cocker, E. D.

Cranfield, C.

L. Fu, A. Jain, C. Cranfield, H. Xie, and M. Gu, J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

Delaney, P.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Denk, W.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

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

Fee, M. S.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

Flusberg, B. A.

Fottner, C.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Fu, L.

Galle, P. R.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Gan, X.

Goetz, M.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Gregor, S.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Gu, M.

Helmchen, F.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

Holtmann, M.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Ilday, F. O.

Jain, A.

L. Fu, A. Jain, C. Cranfield, H. Xie, and M. Gu, J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

Jung, J. C.

B. A. Flusberg, E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, Nat. Methods 12, 941 (2005).
[CrossRef]

B. A. Flusberg, J. C. Jung, E. D. Cocker, E. P. Anderson, and M. J. Schnitzer, Opt. Lett. 30, 2272 (2005).
[CrossRef] [PubMed]

Kanzler, S.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Kiesslich, R.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Klöppel, G.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Ko, T. H.

Lacombe, F.

Lee, D.

Lelek, M.

Li, X.

Louradour, F.

MacDonald, D. J.

Memadathil, B.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Myaing, M. T.

Neurath, M. F.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Piyawattanametha, W.

W. Piyawattanametha, R. P. J. Barretto, T. H. Ko, B. A. Flusberg, E. D. Cocker, H. Ra, D. Lee, and O. Solgaard, Opt. Lett. 31, 2018 (2006).
[CrossRef] [PubMed]

B. A. Flusberg, E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, Nat. Methods 12, 941 (2005).
[CrossRef]

Ra, H.

Schirrmacher, E.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Schirrmacher, R.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Schneider, C.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Schnitzer, M. J.

B. A. Flusberg, E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, Nat. Methods 12, 941 (2005).
[CrossRef]

B. A. Flusberg, J. C. Jung, E. D. Cocker, E. P. Anderson, and M. J. Schnitzer, Opt. Lett. 30, 2272 (2005).
[CrossRef] [PubMed]

Solgaard, O.

Strand, D.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Strickler, J. H.

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

Suran, E.

Tank, D. W.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

Viellerobe, B.

Vieth, M.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Webb, W. W.

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

Weber, M. M.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Weyand, E.

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

Wise, F. W.

Xie, H.

L. Fu, A. Jain, C. Cranfield, H. Xie, and M. Gu, J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

Endoscopy (1)

M. Goetz, C. Fottner, E. Schirrmacher, P. Delaney, S. Gregor, C. Schneider, D. Strand, S. Kanzler, B. Memadathil, E. Weyand, M. Holtmann, R. Schirrmacher, M. M. Weber, M. Anlauf, G. Klöppel, M. Vieth, P. R. Galle, P. Bartenstein, M. F. Neurath, and R. Kiesslich, Endoscopy 39, 350 (2007).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

L. Fu, A. Jain, C. Cranfield, H. Xie, and M. Gu, J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

J. Microsc. (1)

L. Fu and M. Gu, J. Microsc. 226, 195 (2007).
[CrossRef] [PubMed]

Nat. Methods (1)

B. A. Flusberg, E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, Nat. Methods 12, 941 (2005).
[CrossRef]

Neuron (1)

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

Opt. Express (2)

Opt. Lett. (8)

Science (1)

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

Other (1)

M. Gu, Advanced Optical Imaging Theory, Springer Series in Optical Sciences 75 (Springer-Verlag, 2000).

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Figures (4)

Fig. 1
Fig. 1

Schematic of the handheld two-photon fluorescence endoscope. Inset, photo of the probe.

Fig. 2
Fig. 2

(a) Two-photon-excited fluorescence intensity ( I f ) and P N as functions of the grating pair distance for 1 (open triangles), 2 (crosses), and 3 (horizontal bars) m fibers and a 2.9 m fiber connected with the probe (open circles). (b) Log of the two-photon-excited fluorescence intensity ( I f ) versus log of the incident excitation laser power ( P in ) for the 1 m fiber at the optimized grating pair distance.

Fig. 3
Fig. 3

(a) Intensity profile along the center of the two-photon fluorescence image of a Fluoresbrite plain yellow–green (YG) 1 μ m microsphere. (b) Axial response of the two-photon-excited fluorescence signal from a thin layer of DABM dye. (c)–(f) Two-photon-excited fluorescence images of 10 μ m diameter fluorescent microspheres with an axial step of 8 μ m . The image size is 475 μ m × 475 μ m . The excitation wavelength is 800 nm . The excitation power is approximately 15 mW on the sample.

Fig. 4
Fig. 4

(a) Two-photon fluorescence images of a rat kidney surface. (b)–(d) Rat kidney 20, 40, and 60 μ m deep from the surface. The image size is 475 μ m × 475 μ m .

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