Abstract

We report the investigation and implementation of a compact second harmonic generation microscope that uses a single-mode fiber coupler and a double-clad photonic crystal fiber. Second harmonic polarization anisotropy through the fiber-optic microscope systems is quantitatively measured with KTP microcrystals, fish scale and rat tail tendon. It is demonstrated that the polarized second harmonic signals can be excited and collected through the single-mode fiber coupler to analyze the molecular orientations of structural proteins. It has been discovered that a double-clad photonic crystal fiber can preserve the linear polarization in the core, although a depolarization effect is observed in the inner cladding region. The feasibility of polarization anisotropy measurements in fiber-optic second harmonic generation microscopy will benefit the in vivo study of collagen-related diseases with a compact imaging probe.

© 2008 Optical Society of America

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  1. P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
    [CrossRef]
  2. D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
    [CrossRef] [PubMed]
  3. P. J. Campagnola and L. M. Loew, "Second harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat. Biotechnol. 21, 1356-1360 (2003).
    [CrossRef] [PubMed]
  4. L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
    [CrossRef] [PubMed]
  5. P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, "Polarization-modulated second harmonic generation in collagen," Biophys. J. 82, 3330-3342 (2002).
    [CrossRef] [PubMed]
  6. T. Yasui, Y. Tohno, and T. Araki, "Determination of collagen fiber orientation in human tissue by use of polarization measurement of molecular second-harmonic-generation light," Appl. Opt. 43, 2861-2867 (2004).
    [CrossRef] [PubMed]
  7. S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
    [CrossRef] [PubMed]
  8. A. Zoumi, A. Yeh, and B. J. Tromberg, "Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence," Proc. Natl. Acad. Sci. USA 99, 11014-11019 (2002).
    [CrossRef] [PubMed]
  9. W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
    [CrossRef] [PubMed]
  10. S. Yazdanfar, L. H. Laiho, and P. T. C. So, "Interferometric second harmonic generation microscopy," Opt. Express 12, 2739-2745 (2004).
    [CrossRef] [PubMed]
  11. B. E. Applegate, C. Yang, A. M. Rollins, and J. A. Izatt, "Polarization-resolved second-harmonic-generation optical coherence tomography in collagen," Opt. Lett. 29, 2252-2254 (2004).
    [CrossRef] [PubMed]
  12. J. Su, I. V. Tomov, Y. Jiang, and Z. Chen, "High-resolution frequency-domain second-harmonic optical coherence tomography," Appl. Opt. 46, 1770-1775 (2007).
    [CrossRef] [PubMed]
  13. 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]
  14. B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
    [CrossRef] [PubMed]
  15. L. Fu and M. Gu, "Fibre-optic nonlinear optical microscopy and endoscopy," J. Microsc. 226, 195-206 (2007).
    [CrossRef] [PubMed]
  16. D. Bird and M. Gu, "Two-photon fluorescence endoscopy with a micro-optic scanning head," Opt. Lett. 28, 1552-1554 (2003).
    [CrossRef] [PubMed]
  17. 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]
  18. W. GÖbel, J. N. D. Kerr, A. Nimmerjahn, and F. Helmchen, "Miniaturized two-photon microscope based on a flexible coherent fiber bundle and a gradient-index lens objective," Opt. Lett. 29, 2521-2523 (2004).
    [CrossRef] [PubMed]
  19. L. Fu, X. Gan, and M. Gu, "Use of a single-mode fiber coupler for second-harmonic-generation microscopy," Opt. Lett. 30, 385-387 (2005).
    [CrossRef] [PubMed]
  20. 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]
  21. L. Fu, X. Gan, D. Bird, and M. Gu, "Polarisation characteristics of a 1×2 fiber coupler under femtosecond pulsed and continuous wave illumination," Opt. Laser Technol. 37, 494-497 (2005).
    [CrossRef]
  22. Z. Zhu and T. G. Brown, "Polarization properties of supercontinuum spectra generated in birefringent photonic crystal fibers," J. Opt. Soc. Am. B 21, 249-257 (2004).
    [CrossRef]
  23. T. Ritari et al., "Experimental study of polarization properties of highly birefringent photonic crystal fibers," Opt. Express 12, 5931-5939 (2004).
    [CrossRef] [PubMed]
  24. Z. Zhu and T. G. Brown, "Experimental studies of polarization properties of supercontinuum generated in a birefringent photonic crystal fiber," Opt. Express 12, 791-796 (2004).
    [CrossRef] [PubMed]
  25. 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]
  26. L. Fu, A. Jian, C. Cranfield, H. Xie, and M. Gu, "Three-dimensional nonlinear optical endoscopy," J. Biomed. Opt. 12, 040501 (2007).
    [CrossRef] [PubMed]

2007

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

L. Fu, A. Jian, C. Cranfield, H. Xie, and M. Gu, "Three-dimensional nonlinear optical endoscopy," J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

J. Su, I. V. Tomov, Y. Jiang, and Z. Chen, "High-resolution frequency-domain second-harmonic optical coherence tomography," Appl. Opt. 46, 1770-1775 (2007).
[CrossRef] [PubMed]

2006

2005

2004

Z. Zhu and T. G. Brown, "Polarization properties of supercontinuum spectra generated in birefringent photonic crystal fibers," J. Opt. Soc. Am. B 21, 249-257 (2004).
[CrossRef]

Z. Zhu and T. G. Brown, "Experimental studies of polarization properties of supercontinuum generated in a birefringent photonic crystal fiber," Opt. Express 12, 791-796 (2004).
[CrossRef] [PubMed]

T. Yasui, Y. Tohno, and T. Araki, "Determination of collagen fiber orientation in human tissue by use of polarization measurement of molecular second-harmonic-generation light," Appl. Opt. 43, 2861-2867 (2004).
[CrossRef] [PubMed]

S. Yazdanfar, L. H. Laiho, and P. T. C. So, "Interferometric second harmonic generation microscopy," Opt. Express 12, 2739-2745 (2004).
[CrossRef] [PubMed]

B. E. Applegate, C. Yang, A. M. Rollins, and J. A. Izatt, "Polarization-resolved second-harmonic-generation optical coherence tomography in collagen," Opt. Lett. 29, 2252-2254 (2004).
[CrossRef] [PubMed]

W. GÖbel, J. N. D. Kerr, A. Nimmerjahn, and F. Helmchen, "Miniaturized two-photon microscope based on a flexible coherent fiber bundle and a gradient-index lens objective," Opt. Lett. 29, 2521-2523 (2004).
[CrossRef] [PubMed]

T. Ritari et al., "Experimental study of polarization properties of highly birefringent photonic crystal fibers," Opt. Express 12, 5931-5939 (2004).
[CrossRef] [PubMed]

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

2003

W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
[CrossRef] [PubMed]

D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
[CrossRef] [PubMed]

P. J. Campagnola and L. M. Loew, "Second harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat. Biotechnol. 21, 1356-1360 (2003).
[CrossRef] [PubMed]

D. Bird and M. Gu, "Two-photon fluorescence endoscopy with a micro-optic scanning head," Opt. Lett. 28, 1552-1554 (2003).
[CrossRef] [PubMed]

2002

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, "Polarization-modulated second harmonic generation in collagen," Biophys. J. 82, 3330-3342 (2002).
[CrossRef] [PubMed]

A. Zoumi, A. Yeh, and B. J. Tromberg, "Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence," Proc. Natl. Acad. Sci. USA 99, 11014-11019 (2002).
[CrossRef] [PubMed]

2001

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]

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
[CrossRef] [PubMed]

Anderson, E. P.

Applegate, B. E.

Araki, T.

Bird, D.

L. Fu, X. Gan, D. Bird, and M. Gu, "Polarisation characteristics of a 1×2 fiber coupler under femtosecond pulsed and continuous wave illumination," Opt. Laser Technol. 37, 494-497 (2005).
[CrossRef]

D. Bird and M. Gu, "Two-photon fluorescence endoscopy with a micro-optic scanning head," Opt. Lett. 28, 1552-1554 (2003).
[CrossRef] [PubMed]

Blanchard-Desce, M.

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
[CrossRef] [PubMed]

Brown, T. G.

Campagnola, P. J.

P. J. Campagnola and L. M. Loew, "Second harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat. Biotechnol. 21, 1356-1360 (2003).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Celliers, P. M.

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, "Polarization-modulated second harmonic generation in collagen," Biophys. J. 82, 3330-3342 (2002).
[CrossRef] [PubMed]

Charpak, S.

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
[CrossRef] [PubMed]

Chen, S. Y.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

Chen, Y. C.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

Chen, Z.

Chern, G. W.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

Cheung, E. L. M.

B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
[CrossRef] [PubMed]

Christie, R.

W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
[CrossRef] [PubMed]

Chu, S. W.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

Cocker, E. D.

B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
[CrossRef] [PubMed]

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]

Cranfield, C.

Denk, 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]

Dombeck, D. A.

D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
[CrossRef] [PubMed]

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.

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]

B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
[CrossRef] [PubMed]

Fu, L.

L. Fu, A. Jian, C. Cranfield, H. Xie, and M. Gu, "Three-dimensional nonlinear optical endoscopy," J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

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

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]

L. Fu, X. Gan, and M. Gu, "Use of a single-mode fiber coupler for second-harmonic-generation microscopy," Opt. Lett. 30, 385-387 (2005).
[CrossRef] [PubMed]

L. Fu, X. Gan, D. Bird, and M. Gu, "Polarisation characteristics of a 1×2 fiber coupler under femtosecond pulsed and continuous wave illumination," Opt. Laser Technol. 37, 494-497 (2005).
[CrossRef]

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]

Gan, X.

Gu, M.

Helmchen, F.

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]

Hoppe, P. E.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Hyman, B. T.

W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
[CrossRef] [PubMed]

D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
[CrossRef] [PubMed]

Ingelsson, M.

D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
[CrossRef] [PubMed]

Izatt, J. A.

Jain, A.

Jian, A.

L. Fu, A. Jian, C. Cranfield, H. Xie, and M. Gu, "Three-dimensional nonlinear optical endoscopy," J. Biomed. Opt. 12, 040501 (2007).
[CrossRef] [PubMed]

Jiang, Y.

Jung, J. C.

B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
[CrossRef] [PubMed]

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]

Kasischke, K. A.

D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
[CrossRef] [PubMed]

Laiho, L. H.

Lin, B. L.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

Loew, L. M.

P. J. Campagnola and L. M. Loew, "Second harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat. Biotechnol. 21, 1356-1360 (2003).
[CrossRef] [PubMed]

Malone, C. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Mertz, J.

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
[CrossRef] [PubMed]

Millard, A. C.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Mohler, W. A.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Moreaux, L.

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
[CrossRef] [PubMed]

Nikitin, A. Y.

W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
[CrossRef] [PubMed]

Piyawattanametha, W.

B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
[CrossRef] [PubMed]

Reiser, K. M.

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, "Polarization-modulated second harmonic generation in collagen," Biophys. J. 82, 3330-3342 (2002).
[CrossRef] [PubMed]

Ritari, T.

Rollins, A. M.

Rubenchik, A. M.

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, "Polarization-modulated second harmonic generation in collagen," Biophys. J. 82, 3330-3342 (2002).
[CrossRef] [PubMed]

Sandre, O.

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
[CrossRef] [PubMed]

Schnitzer, M. J.

B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
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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).
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P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, "Polarization-modulated second harmonic generation in collagen," Biophys. J. 82, 3330-3342 (2002).
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S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[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]

Terasaki, M.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

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A. Zoumi, A. Yeh, and B. J. Tromberg, "Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence," Proc. Natl. Acad. Sci. USA 99, 11014-11019 (2002).
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S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
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D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
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D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
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W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
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A. Zoumi, A. Yeh, and B. J. Tromberg, "Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence," Proc. Natl. Acad. Sci. USA 99, 11014-11019 (2002).
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W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
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A. Zoumi, A. Yeh, and B. J. Tromberg, "Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence," Proc. Natl. Acad. Sci. USA 99, 11014-11019 (2002).
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Appl. Opt.

Biophys. J.

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, "Coherent scattering in multi-harmonic light microscopy," Biophys. J. 80, 1568-1574 (2001).
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[CrossRef] [PubMed]

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x(2)/x(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
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L. Fu, A. Jian, C. Cranfield, H. Xie, and M. Gu, "Three-dimensional nonlinear optical endoscopy," J. Biomed. Opt. 12, 040501 (2007).
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L. Fu and M. Gu, "Fibre-optic nonlinear optical microscopy and endoscopy," J. Microsc. 226, 195-206 (2007).
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Nat. Biotechnol.

P. J. Campagnola and L. M. Loew, "Second harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat. Biotechnol. 21, 1356-1360 (2003).
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Nat. Methods

B. A. Flusberg and E. D. Cocker, W. Piyawattanametha, J. C. Jung, E. L. M. Cheung, and M. J. Schnitzer, "Fiber-optic fluorescence imaging," Nat. Methods 2, 941-950 (2005).
[CrossRef] [PubMed]

Neuron.

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. Express

Opt. Laser Technol.

L. Fu, X. Gan, D. Bird, and M. Gu, "Polarisation characteristics of a 1×2 fiber coupler under femtosecond pulsed and continuous wave illumination," Opt. Laser Technol. 37, 494-497 (2005).
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Opt. Lett.

Proc. Natl. Acad. Sci. USA

D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy," Proc. Natl. Acad. Sci. USA 100, 7081-7086 (2003).
[CrossRef] [PubMed]

A. Zoumi, A. Yeh, and B. J. Tromberg, "Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence," Proc. Natl. Acad. Sci. USA 99, 11014-11019 (2002).
[CrossRef] [PubMed]

W. E. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation," Proc. Natl. Acad. Sci. USA 100, 7075-7080 (2003).
[CrossRef] [PubMed]

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Figures (5)

Fig. 1.
Fig. 1.

Schematic diagram of the SHG microscope based on (a) a single-mode fiber coupler and (b) a double-clad PCF for polarization anisotropy measurement. ND: Neutral density filter, GTP: Glan Thompson polarizer, O1 and O2: microscope objectives, O3: Olympus 40×/0.85NA imaging objective.

Fig. 2.
Fig. 2.

SHG polarization anisotropy measurement with the KTP microcrystals. SHG images are obtained with orthogonal polarization orientations of the analyzer in a standard laser scanning microscope (a,b) and a SHG microscope using a single-mode fiber coupler (c,d), respectively. (e) Dependence of the SHG intensity on the rotation angle of the analyzer in a laser scanning (nonfiber) microscope and a single-mode fiber coupler-based microscope, where the results fit a cos2 θ function. Each image has a dimension of 30 µm×3 0 µm.

Fig. 3
Fig. 3

SHG polarization anisotropy measurement with a fish scale in a SHG microscope using a single-mode fiber coupler. (a),(b) SHG images obtained with orthogonal polarization orientations of analyzer. Scale bar is 20 µm. (c) Image of the anisotropy parameter derived from (a) and (b). Color scale varies from blue for -0.5 to red for 1.0.

Fig. 4.
Fig. 4.

Degree of polarization of the laser beam delivered by (a) the core/inner cladding region and (b) the central core of the double-clad PCF as a function of the linear polarization angle of the incident beam at wavelength 800 nm.

Fig. 5.
Fig. 5.

SHG polarization anisotropy measurement with (a) a fish scale and (b) a rat tail tendon in a SHG microscope using a double-clad PCF. Each set includes SHG images that are obtained without an analyzer and with orthogonal polarization orientations of the analyzer, and the anisotropy parameter image. Scale bars are 10 µm. Color scale varies from blue for -0.5 to red for 1.0.

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