Abstract

We present a rigid miniaturized optical system block fiber-optic two-photon endoscope based on a compact two-axis piezo scanner system and a miniature high (0.65) NA GRIN lens objective. The optical system is scanned as a whole by a piezo scanner allowing always an on-axis beam irradiation of the optical system. A photonic crystal fiber is used for excitation and ultrashort laser pulses can be delivered with typical power up to 100mW at 800nm. Two-photon fluorescence signal is collected by the use of a multimode fiber. Lateral resolution values for the system were experimentally measured to be 0.67μm vertically and 1.08μm horizontally. Axial resolution was found to be 5.8μm. The endoscope is highly flexible and controllable in terms of time acquisition, resolution, and magnification. Fluorescence images were acquired over a 420μm×420μm field of view. Results presented here demonstrate the ability of the system to resolve subcellular details and the potential of the technology for in vivo applications.

© 2009 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning microscopy,” Science 248, 73-76 (1990).
    [CrossRef] [PubMed]
  2. K. König, “Review: multiphoton microscopy in life sciences,” J. Microsc. 200, 83-104 (2000).
    [CrossRef] [PubMed]
  3. K. König and I. Riemann, “High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution,” J. Biomed. Opt. 8, 432-439(2003).
    [CrossRef] [PubMed]
  4. R. Richards-Kortum and E. Sevick-Muraca, “Quantitative optical spectroscopy for tissue diagnosis,” Annu. Rev. Phys. Chem. 47, 555-606 (1996).
    [CrossRef] [PubMed]
  5. W. A. Reed, M. F. Yan, and M. J. Schnitzer, “Gradient-index fiber-optic microprobes for minimally invasive in vivo low-coherence interferometry,” Opt Lett 27, 1794-1796 (2002).
    [CrossRef]
  6. J. C. Jung and M. J. Chnitzer, “Multiphoton endoscopy,” Opt. Lett. 28, 902-904 (2003).
    [CrossRef] [PubMed]
  7. M. T. Myaing, D. J. MacDonald, and X. Li, “Fiber-optic scanning two-photon fluorescence endoscope,” Opt. Lett. 31, 1076-1078 (2006).
    [CrossRef] [PubMed]
  8. C. J. Engelbrecht, R. S. Johnston, E. J. Seibel, and F. Helmchen, “Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo,” Opt. Express 16, 5556-5564 (2008).
    [CrossRef] [PubMed]
  9. B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
    [CrossRef]
  10. S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
    [CrossRef]
  11. K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
    [CrossRef] [PubMed]
  12. F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope: high-resolution brain imaging in freely moving animals,” Neuron 31, 903-912(2001).
    [CrossRef] [PubMed]
  13. E. J. Seibel and Q. Y. L. Smithwick, “Unique features of optical scanning, single fiber endoscopy,” Lasers Surg. Med. 30, 177-183 (2002).
    [CrossRef] [PubMed]
  14. L. Fu, X. Gan, and M. Gu, “Nonlinear optical microscopy based on double-clad photonic crystal fibers,” Opt. Express 13, 5528-5534 (2005).
    [CrossRef] [PubMed]
  15. B. A. Flusberg, J. C. Jung, E. D. Cocker, E. P. Anderson, and M. J. Schnitzer, “In vivo brain imaging using a portable 3.9 gram two-photon fluorescence microendoscope,” Opt. Lett. 30, 2272-2274 (2005).
    [CrossRef] [PubMed]
  16. W. Piyawattanametha, R. P. J. Barretto, T. H. Ko, B. A. Flusberg, E. D. Cocker, H. Ra, D. Lee, O. Solgaard, and M. J. Schnitzer, “Fast-scanning two-photon fluorescence imaging based on a microelectromechanical systems two-dimensional scanning mirror,” Opt. Lett. 31, 2018-2020(2006).
    [CrossRef] [PubMed]
  17. J. Sawinski and W. Denk, “Miniature random-access fiber scanner for in vivo multiphoton imaging,” J. Appl. Phys. 102, 034701 (2007).
    [CrossRef]
  18. E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
    [CrossRef]
  19. L. Fu, A. Jain, H. Xie, C. Cranfield, and M. Gu, “Nonlinear optical endoscopy based on a double-clad photonic crystal fiber and a MEMS mirror,” Opt. Express 14, 1027-1032(2006).
    [CrossRef] [PubMed]
  20. L. Fu and M. Gu, “Fibre-optic nonlinear optical microscopy and endoscopy,” J. Microsc. (Oxford) 226, 195-206 (2007).
    [CrossRef]
  21. C. S. Croix, W. R. Zipfel, and S. C. Watkins, “Potential solutions for confocal imaging of living animals,” BioTechniques 43, S14-S19 (2007).
    [CrossRef]
  22. I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
    [CrossRef]
  23. C. L. Hoy, N. J. Durr, P. Chen, W. Piyawattanametha, H. Ra, O. Solgaard, and A. Ben-Yakar, “Miniaturized probe for femtosecond laser microsurgery and two-photon imaging,” Opt. Express 16, 9996-10005 (2008).
    [CrossRef] [PubMed]
  24. A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
    [CrossRef]

2008 (3)

2007 (7)

J. Sawinski and W. Denk, “Miniature random-access fiber scanner for in vivo multiphoton imaging,” J. Appl. Phys. 102, 034701 (2007).
[CrossRef]

E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
[CrossRef]

L. Fu and M. Gu, “Fibre-optic nonlinear optical microscopy and endoscopy,” J. Microsc. (Oxford) 226, 195-206 (2007).
[CrossRef]

C. S. Croix, W. R. Zipfel, and S. C. Watkins, “Potential solutions for confocal imaging of living animals,” BioTechniques 43, S14-S19 (2007).
[CrossRef]

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

2006 (3)

2005 (2)

2003 (2)

J. C. Jung and M. J. Chnitzer, “Multiphoton endoscopy,” Opt. Lett. 28, 902-904 (2003).
[CrossRef] [PubMed]

K. König and I. Riemann, “High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution,” J. Biomed. Opt. 8, 432-439(2003).
[CrossRef] [PubMed]

2002 (2)

W. A. Reed, M. F. Yan, and M. J. Schnitzer, “Gradient-index fiber-optic microprobes for minimally invasive in vivo low-coherence interferometry,” Opt Lett 27, 1794-1796 (2002).
[CrossRef]

E. J. Seibel and Q. Y. L. Smithwick, “Unique features of optical scanning, single fiber endoscopy,” Lasers Surg. Med. 30, 177-183 (2002).
[CrossRef] [PubMed]

2001 (1)

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope: high-resolution brain imaging in freely moving animals,” Neuron 31, 903-912(2001).
[CrossRef] [PubMed]

2000 (1)

K. König, “Review: multiphoton microscopy in life sciences,” J. Microsc. 200, 83-104 (2000).
[CrossRef] [PubMed]

1999 (1)

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

1996 (1)

R. Richards-Kortum and E. Sevick-Muraca, “Quantitative optical spectroscopy for tissue diagnosis,” Annu. Rev. Phys. Chem. 47, 555-606 (1996).
[CrossRef] [PubMed]

1990 (1)

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

Abicht, H.

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

Anderson, E. P.

Barretto, R. P. J.

Beige, H.

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

Ben-Yakar, A.

Brown, C. M.

E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
[CrossRef]

Bückle, R.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

Chen, P.

Chnitzer, M. J.

Cocker, E. D.

Cranfield, C.

Croix, C. S.

C. S. Croix, W. R. Zipfel, and S. C. Watkins, “Potential solutions for confocal imaging of living animals,” BioTechniques 43, S14-S19 (2007).
[CrossRef]

Denk, W.

J. Sawinski and W. Denk, “Miniature random-access fiber scanner for in vivo multiphoton imaging,” J. Appl. Phys. 102, 034701 (2007).
[CrossRef]

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope: high-resolution brain imaging in freely moving animals,” Neuron 31, 903-912(2001).
[CrossRef] [PubMed]

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

Dominitz, J. A.

E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
[CrossRef]

Durr, N. J.

Ehlers, A.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

Engelbrecht, C. J.

Fee, M. S.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope: high-resolution brain imaging in freely moving animals,” Neuron 31, 903-912(2001).
[CrossRef] [PubMed]

Flusberg, B. A.

Fu, L.

Gan, X.

Gu, M.

Hauke, T.

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

Helmchen, F.

C. J. Engelbrecht, R. S. Johnston, E. J. Seibel, and F. Helmchen, “Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo,” Opt. Express 16, 5556-5564 (2008).
[CrossRef] [PubMed]

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope: high-resolution brain imaging in freely moving animals,” Neuron 31, 903-912(2001).
[CrossRef] [PubMed]

Hoy, C. L.

Jain, A.

Johnston, R. S.

C. J. Engelbrecht, R. S. Johnston, E. J. Seibel, and F. Helmchen, “Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo,” Opt. Express 16, 5556-5564 (2008).
[CrossRef] [PubMed]

E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
[CrossRef]

Jung, J. C.

Kaatz, M.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

Kimmey, M. B.

E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
[CrossRef]

Ko, T. H.

König, K.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

K. König and I. Riemann, “High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution,” J. Biomed. Opt. 8, 432-439(2003).
[CrossRef] [PubMed]

K. König, “Review: multiphoton microscopy in life sciences,” J. Microsc. 200, 83-104 (2000).
[CrossRef] [PubMed]

Kouvatov, A.

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

Kraeplin, A.

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

Langhammer, H. T.

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

Le Harzic, R.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

Lee, D.

Li, X.

MacDonald, D. J.

Messerschmidt, B.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

Messerscmidt, B.

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

Myaing, M. T.

Piyawattanametha, W.

Ra, H.

Reed, W. A.

W. A. Reed, M. F. Yan, and M. J. Schnitzer, “Gradient-index fiber-optic microprobes for minimally invasive in vivo low-coherence interferometry,” Opt Lett 27, 1794-1796 (2002).
[CrossRef]

Richards-Kortum, R.

R. Richards-Kortum and E. Sevick-Muraca, “Quantitative optical spectroscopy for tissue diagnosis,” Annu. Rev. Phys. Chem. 47, 555-606 (1996).
[CrossRef] [PubMed]

Riemann, I.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

K. König and I. Riemann, “High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution,” J. Biomed. Opt. 8, 432-439(2003).
[CrossRef] [PubMed]

Sauer, D.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

Sawinski, J.

J. Sawinski and W. Denk, “Miniature random-access fiber scanner for in vivo multiphoton imaging,” J. Appl. Phys. 102, 034701 (2007).
[CrossRef]

Schenkl, S.

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

Schnitzer, M. J.

Seibel, E. J.

C. J. Engelbrecht, R. S. Johnston, E. J. Seibel, and F. Helmchen, “Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo,” Opt. Express 16, 5556-5564 (2008).
[CrossRef] [PubMed]

E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
[CrossRef]

E. J. Seibel and Q. Y. L. Smithwick, “Unique features of optical scanning, single fiber endoscopy,” Lasers Surg. Med. 30, 177-183 (2002).
[CrossRef] [PubMed]

Seifert, W.

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

Sevick-Muraca, E.

R. Richards-Kortum and E. Sevick-Muraca, “Quantitative optical spectroscopy for tissue diagnosis,” Annu. Rev. Phys. Chem. 47, 555-606 (1996).
[CrossRef] [PubMed]

Smithwick, Q. Y. L.

E. J. Seibel and Q. Y. L. Smithwick, “Unique features of optical scanning, single fiber endoscopy,” Lasers Surg. Med. 30, 177-183 (2002).
[CrossRef] [PubMed]

Solgaard, O.

Stark, M.

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

Steinhausen, R.

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

Strickler, J. H.

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

Tank, D. W.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope: high-resolution brain imaging in freely moving animals,” Neuron 31, 903-912(2001).
[CrossRef] [PubMed]

Tchernook, A.

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

Watkins, S. C.

C. S. Croix, W. R. Zipfel, and S. C. Watkins, “Potential solutions for confocal imaging of living animals,” BioTechniques 43, S14-S19 (2007).
[CrossRef]

Webb, W. W.

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

Xie, H.

Yan, M. F.

W. A. Reed, M. F. Yan, and M. J. Schnitzer, “Gradient-index fiber-optic microprobes for minimally invasive in vivo low-coherence interferometry,” Opt Lett 27, 1794-1796 (2002).
[CrossRef]

Zipfel, W. R.

C. S. Croix, W. R. Zipfel, and S. C. Watkins, “Potential solutions for confocal imaging of living animals,” BioTechniques 43, S14-S19 (2007).
[CrossRef]

Annu. Rev. Phys. Chem. (1)

R. Richards-Kortum and E. Sevick-Muraca, “Quantitative optical spectroscopy for tissue diagnosis,” Annu. Rev. Phys. Chem. 47, 555-606 (1996).
[CrossRef] [PubMed]

BioTechniques (1)

C. S. Croix, W. R. Zipfel, and S. C. Watkins, “Potential solutions for confocal imaging of living animals,” BioTechniques 43, S14-S19 (2007).
[CrossRef]

Gastrointest. Endosc. (1)

E. J. Seibel, R. S. Johnston, C. M. Brown, J. A. Dominitz, and M. B. Kimmey, “Novel ultrathin scanning fiber endoscope for cholangioscopy and pancreatoscopy,” Gastrointest. Endosc. 65, Ab125-Ab125 (2007).
[CrossRef]

J. Appl. Phys. (1)

J. Sawinski and W. Denk, “Miniature random-access fiber scanner for in vivo multiphoton imaging,” J. Appl. Phys. 102, 034701 (2007).
[CrossRef]

J. Biomed. Opt. (1)

K. König and I. Riemann, “High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution,” J. Biomed. Opt. 8, 432-439(2003).
[CrossRef] [PubMed]

J. Eur. Ceram. Soc. (1)

A. Kouvatov, R. Steinhausen, W. Seifert, T. Hauke, H. T. Langhammer, H. Beige, and H. Abicht, “Comparison between bimorphic and polymorphic bending devices,” J. Eur. Ceram. Soc. 19, 1153-1156 (1999).
[CrossRef]

J. Microsc. (1)

K. König, “Review: multiphoton microscopy in life sciences,” J. Microsc. 200, 83-104 (2000).
[CrossRef] [PubMed]

J. Microsc. (Oxford) (1)

L. Fu and M. Gu, “Fibre-optic nonlinear optical microscopy and endoscopy,” J. Microsc. (Oxford) 226, 195-206 (2007).
[CrossRef]

Lasers Surg. Med. (1)

E. J. Seibel and Q. Y. L. Smithwick, “Unique features of optical scanning, single fiber endoscopy,” Lasers Surg. Med. 30, 177-183 (2002).
[CrossRef] [PubMed]

Microsc. Res. Tech. (1)

K. König, A. Ehlers, I. Riemann, S. Schenkl, R. Bückle, and M. Kaatz, “Clinical two-photon microendoscopy,” Microsc. Res. Tech. 70, 398-402 (2007).
[CrossRef] [PubMed]

Neuron (1)

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope: high-resolution brain imaging in freely moving animals,” Neuron 31, 903-912(2001).
[CrossRef] [PubMed]

Opt Lett (1)

W. A. Reed, M. F. Yan, and M. J. Schnitzer, “Gradient-index fiber-optic microprobes for minimally invasive in vivo low-coherence interferometry,” Opt Lett 27, 1794-1796 (2002).
[CrossRef]

Opt. Express (4)

Opt. Lett. (4)

Proc. SPIE (2)

I. Riemann, S. Schenkl, R. Le Harzic, D. Sauer, A. Ehlers, B. Messerschmidt, M. Kaatz, R. Bückle, and K. König, “Two-photon imaging using a flexible endoscope,” Proc. SPIE 6851, 68510B (2008).
[CrossRef]

S. Schenkl, A. Ehlers, R. Le Harzic, M. Stark, I. Riemann, B. Messerscmidt, M. Kaatz, and K. König, “Rigid and high NA multiphoton fluorescence GRIN-endoscopes,” Proc. SPIE 6631, 66310Q (2007).
[CrossRef]

Proc.SPIE (1)

B. Messerschmidt, A. Kraeplin, S. Schenkl, I. Riemann, M. Stark, A. Ehlers, A. Tchernook, R. Le Harzic, and K. König, “Novel concept of GRIN optical systems for high resolution microendoscopy: Part 1. Physical aspects,” Proc.SPIE 6432, 643202 (2007).
[CrossRef]

Science (1)

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

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 (7)

Fig. 1
Fig. 1

Schematic representation of the compact piezo-scanner two-photon endoscope. A photograph of the endoscope head is given in the inset. PMT, photomultiplier tube; APD, avalanche photodiode.

Fig. 2
Fig. 2

Details of the mechanical and optical body of the endoscope.

Fig. 3
Fig. 3

Photographs of the piezo scanner based on trimorph actuators.

Fig. 4
Fig. 4

Two-photon image of mixed preparation of 0.5 and 2 μm microspheres at (a) 0%, (b) 50%, and (c) 80% zoom factor.

Fig. 5
Fig. 5

Two-photon images of a grid target with line widths of 1.5 μm and spaced every 13 μm . The grid has been filled with Rhodamine 123 for fluorescence detection. (a) focus shift forward (b) focus at the center, (c) focus shift rear.

Fig. 6
Fig. 6

Fluorescence intensity profiles of a 0.5 μm example microsphere along the lateral and axial dimension are shown. The solid curves are Gaussian-shaped curves fitted to the data points.

Fig. 7
Fig. 7

Two-photon image of labeled mitochondria in a fixed CHO cell. Laser parameters: λ = 800 nm , P = 60 mW . Image parameters: frame rate, 80 s ; number of pixels, 512 × 512 .

Metrics