Abstract

In this study, we propose a novel, minimally-invasive multimodal optical imaging method which combines multiphoton and reflected confocal microscopy for characterizing three-dimensional phase-separated microstructure of polymeric nylon/chitosan blends. The multimodal image acquisition was performed on a Zeiss LSM 510 inverted microscope system using a ti-sapphire laser source. Differences in nonlinear optical signals between individual homopolymers were used to characterize the phase-separation phenomenon within the polymeric blends. We also used the reflected confocal signals for defining the interfacial boundaries of different refractive indices. Our work demonstrates that the proposed multimodal imaging modality can be used to provide the necessary microstructural information for characterizing the degrees of phase separation within polymeric blends.

© 2008 Optical Society of America

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  1. R. A. Kudva, H. Keskkula, and D. R. Paul, "Compatibilization of nylon 6/ABS blends using glycidyl methacrylate/methyl methacrylate copolymers," Polymer 39, 2447-2460 (1998).
    [CrossRef]
  2. J. Kressler, N. Higashida, T. Inoue, W. Heckmann, and F. Seitz, "Study of Polymer-Polymer Interfaces: A Comparison of Ellipsometric and TEM Data of PMMA/PS and PMMA/SAN Systems," Macromolecules 26, 2090-2094 (1993).
    [CrossRef]
  3. J. H. Wang, C. W. Wei, H. C. Liu, and T. H. Young, "Behavior of MG-63 cells on nylon/chitosan-blended membranes," J. Biomed. Mater. Res. A. 64, 606-615 (2003).
    [CrossRef] [PubMed]
  4. S. J. Lin, W. C. Hsiao, S. H. Jee, H. S. Yu, T. F. Tsai, J. Y. Lai, and T. H. Young, "Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes," Biomaterials 27, 5079-5088 (2006).
    [CrossRef] [PubMed]
  5. P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).
  6. B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
    [CrossRef]
  7. A. Gupper, P. Wilhelm, M. Schmied, and E. Ingolic. "Morphology of a PA/PTFE blend studied by Raman imaging," Macromol. Symp. 184, 275-286 (2002).
    [CrossRef]
  8. A. Chris, X. Gu, D. B. Chase, and S. J. Stranick. "Near-Field Infrared Imaging and Spectroscopy of a Thin Film Polystyrene/Poly(ethyl acrylate) Blend," Appl. Spectrosc. 58, 257-263 (2004).
    [CrossRef]
  9. H. Jinnai, Y. Nishikawa, T. Koga, and T. Hashimoto, "Direct Observation of Three-dimensional Bicontinuous Structure Developed via Spinodal Decomposition," Macromolecules 28, 4782-4784 (1995).
    [CrossRef]
  10. A. E. Ribbe, "Laser scanning confocal microscopy in polymer science," Trends Polym. Sci. 5, 333-337 (1997).
  11. H. Jinnai, H. Kitagishi, K. Hamano, Y. Nishikawa, and Takahashi, "Effect of confinement on phase-separation processes in a polymer blend observed by laser scanning confocal microscopy," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 67, 021801 (2003).
    [CrossRef] [PubMed]
  12. A. E. Ribbe, M. Hayashi, M. Weber, and T. Hashimoto, "Identification of the morphology of a thin film by fluorescence laser-scanning confocal microscopy," Polymer. 39, 7149-7151(1998).
    [CrossRef]
  13. K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
    [CrossRef] [PubMed]
  14. J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
    [CrossRef] [PubMed]
  15. 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," P. Natl. Acad. Sci. U.S.A. 99, 11014-11019 (2002).
    [CrossRef]
  16. H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
    [CrossRef]
  17. I. Constantinidis, C. L. Stabler, R. Long, and A. Sambanis, "Noninvasive monitoring of a retrievable bioartificial pancreas in vivo," Ann. N. Y. Acad. Sci. 961, 298-301 (2002).
    [CrossRef] [PubMed]
  18. Y. W. Wang, Q. Wu, J. C. Chen, and G. Q. Chen, "Evaluation of three-dimensional scaffolds made of blends of hydroxyapatite and poly(3-hydroxybutyrate-co- 3-hydroxyhexanoate) for bone reconstruction," Biomaterials 26, 899-904 (2005).
    [CrossRef]
  19. D. M. Burland, R. D. Miller, and C. A. Walsh. "Second-order nonlinearity in poled-polymer systems," Chem. Rev. 94, 31-75 (1994).
    [CrossRef]
  20. J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
    [CrossRef]
  21. S. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys J. 90, 693-703 (2006).
    [CrossRef]
  22. N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
    [CrossRef] [PubMed]
  23. W. Tan, A. Sendermir-Urkmez, L. J. Fahrmer, R. Jamison, and D. Leckband, "Structual and functional optical imaging of three-dimensional engineered tissue development," Tissue. Eng. 10, 1747-1757 (2004).
    [CrossRef]
  24. F. L. Mi, "Synthesis and characterization of a novel chitosan-gelatin bioconjugate with fluorescence emission," Biomacromolecules 6, 975-987 (2005).
    [CrossRef] [PubMed]
  25. A. Dufresne, J. Y. Cavaille, D. Dupeyre, M. Garcia-Ramirez, and J. Romero, "Morphology, phase continuity and mechanical behavior of polyamide 6/chitosan blends," Polymer. 40, 1657-1666 (1999).
    [CrossRef]
  26. A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
    [CrossRef] [PubMed]
  27. A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
    [CrossRef]
  28. S. Tang, T. B. Krasieva, Z. Chen, B. J. Tromberg, "Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source," J. Biomed. Opt. 11, 020502 (2006).
    [CrossRef] [PubMed]

2007 (1)

J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
[CrossRef]

2006 (5)

S. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys J. 90, 693-703 (2006).
[CrossRef]

S. Tang, T. B. Krasieva, Z. Chen, B. J. Tromberg, "Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source," J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

S. J. Lin, W. C. Hsiao, S. H. Jee, H. S. Yu, T. F. Tsai, J. Y. Lai, and T. H. Young, "Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes," Biomaterials 27, 5079-5088 (2006).
[CrossRef] [PubMed]

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

2005 (3)

N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
[CrossRef] [PubMed]

Y. W. Wang, Q. Wu, J. C. Chen, and G. Q. Chen, "Evaluation of three-dimensional scaffolds made of blends of hydroxyapatite and poly(3-hydroxybutyrate-co- 3-hydroxyhexanoate) for bone reconstruction," Biomaterials 26, 899-904 (2005).
[CrossRef]

F. L. Mi, "Synthesis and characterization of a novel chitosan-gelatin bioconjugate with fluorescence emission," Biomacromolecules 6, 975-987 (2005).
[CrossRef] [PubMed]

2004 (3)

W. Tan, A. Sendermir-Urkmez, L. J. Fahrmer, R. Jamison, and D. Leckband, "Structual and functional optical imaging of three-dimensional engineered tissue development," Tissue. Eng. 10, 1747-1757 (2004).
[CrossRef]

A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
[CrossRef] [PubMed]

A. Chris, X. Gu, D. B. Chase, and S. J. Stranick. "Near-Field Infrared Imaging and Spectroscopy of a Thin Film Polystyrene/Poly(ethyl acrylate) Blend," Appl. Spectrosc. 58, 257-263 (2004).
[CrossRef]

2003 (2)

J. H. Wang, C. W. Wei, H. C. Liu, and T. H. Young, "Behavior of MG-63 cells on nylon/chitosan-blended membranes," J. Biomed. Mater. Res. A. 64, 606-615 (2003).
[CrossRef] [PubMed]

H. Jinnai, H. Kitagishi, K. Hamano, Y. Nishikawa, and Takahashi, "Effect of confinement on phase-separation processes in a polymer blend observed by laser scanning confocal microscopy," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 67, 021801 (2003).
[CrossRef] [PubMed]

2002 (4)

K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
[CrossRef] [PubMed]

I. Constantinidis, C. L. Stabler, R. Long, and A. Sambanis, "Noninvasive monitoring of a retrievable bioartificial pancreas in vivo," Ann. N. Y. Acad. Sci. 961, 298-301 (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," P. Natl. Acad. Sci. U.S.A. 99, 11014-11019 (2002).
[CrossRef]

A. Gupper, P. Wilhelm, M. Schmied, and E. Ingolic. "Morphology of a PA/PTFE blend studied by Raman imaging," Macromol. Symp. 184, 275-286 (2002).
[CrossRef]

2001 (1)

A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
[CrossRef]

2000 (1)

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

1999 (1)

A. Dufresne, J. Y. Cavaille, D. Dupeyre, M. Garcia-Ramirez, and J. Romero, "Morphology, phase continuity and mechanical behavior of polyamide 6/chitosan blends," Polymer. 40, 1657-1666 (1999).
[CrossRef]

1998 (2)

R. A. Kudva, H. Keskkula, and D. R. Paul, "Compatibilization of nylon 6/ABS blends using glycidyl methacrylate/methyl methacrylate copolymers," Polymer 39, 2447-2460 (1998).
[CrossRef]

A. E. Ribbe, M. Hayashi, M. Weber, and T. Hashimoto, "Identification of the morphology of a thin film by fluorescence laser-scanning confocal microscopy," Polymer. 39, 7149-7151(1998).
[CrossRef]

1997 (1)

A. E. Ribbe, "Laser scanning confocal microscopy in polymer science," Trends Polym. Sci. 5, 333-337 (1997).

1996 (1)

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

1995 (1)

H. Jinnai, Y. Nishikawa, T. Koga, and T. Hashimoto, "Direct Observation of Three-dimensional Bicontinuous Structure Developed via Spinodal Decomposition," Macromolecules 28, 4782-4784 (1995).
[CrossRef]

1994 (1)

D. M. Burland, R. D. Miller, and C. A. Walsh. "Second-order nonlinearity in poled-polymer systems," Chem. Rev. 94, 31-75 (1994).
[CrossRef]

1993 (1)

J. Kressler, N. Higashida, T. Inoue, W. Heckmann, and F. Seitz, "Study of Polymer-Polymer Interfaces: A Comparison of Ellipsometric and TEM Data of PMMA/PS and PMMA/SAN Systems," Macromolecules 26, 2090-2094 (1993).
[CrossRef]

Achalla, P.

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

Bao, J.

J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
[CrossRef]

Baselga, J.

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

Berezney, R.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Bhawalkar, J. D.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Bravo, J.

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

Burland, D. M.

D. M. Burland, R. D. Miller, and C. A. Walsh. "Second-order nonlinearity in poled-polymer systems," Chem. Rev. 94, 31-75 (1994).
[CrossRef]

Campagnola, P. J.

S. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys J. 90, 693-703 (2006).
[CrossRef]

Cavaille, J. Y.

A. Dufresne, J. Y. Cavaille, D. Dupeyre, M. Garcia-Ramirez, and J. Romero, "Morphology, phase continuity and mechanical behavior of polyamide 6/chitosan blends," Polymer. 40, 1657-1666 (1999).
[CrossRef]

Chase, D. B.

Chen, G. Q.

J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
[CrossRef]

Y. W. Wang, Q. Wu, J. C. Chen, and G. Q. Chen, "Evaluation of three-dimensional scaffolds made of blends of hydroxyapatite and poly(3-hydroxybutyrate-co- 3-hydroxyhexanoate) for bone reconstruction," Biomaterials 26, 899-904 (2005).
[CrossRef]

Chen, H. C.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Chen, J. C.

Y. W. Wang, Q. Wu, J. C. Chen, and G. Q. Chen, "Evaluation of three-dimensional scaffolds made of blends of hydroxyapatite and poly(3-hydroxybutyrate-co- 3-hydroxyhexanoate) for bone reconstruction," Biomaterials 26, 899-904 (2005).
[CrossRef]

Chen, Z.

S. Tang, T. B. Krasieva, Z. Chen, B. J. Tromberg, "Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source," J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
[CrossRef] [PubMed]

Cheng, P. C.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Chiou, L. L.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Chris, A.

Collier, T.

K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
[CrossRef] [PubMed]

Constantinidis, I.

I. Constantinidis, C. L. Stabler, R. Long, and A. Sambanis, "Noninvasive monitoring of a retrievable bioartificial pancreas in vivo," Ann. N. Y. Acad. Sci. 961, 298-301 (2002).
[CrossRef] [PubMed]

Descour, M.

K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
[CrossRef] [PubMed]

Diaspro, A.

A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
[CrossRef]

Dong, C. Y.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Dufresne, A.

A. Dufresne, J. Y. Cavaille, D. Dupeyre, M. Garcia-Ramirez, and J. Romero, "Morphology, phase continuity and mechanical behavior of polyamide 6/chitosan blends," Polymer. 40, 1657-1666 (1999).
[CrossRef]

Dupeyre, D.

A. Dufresne, J. Y. Cavaille, D. Dupeyre, M. Garcia-Ramirez, and J. Romero, "Morphology, phase continuity and mechanical behavior of polyamide 6/chitosan blends," Polymer. 40, 1657-1666 (1999).
[CrossRef]

Esnault, P.

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

Esteban, I.

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

Fahrmer, L. J.

W. Tan, A. Sendermir-Urkmez, L. J. Fahrmer, R. Jamison, and D. Leckband, "Structual and functional optical imaging of three-dimensional engineered tissue development," Tissue. Eng. 10, 1747-1757 (2004).
[CrossRef]

Federici, F.

A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
[CrossRef]

Follen, M.

K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
[CrossRef] [PubMed]

Gao, L.

J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
[CrossRef]

Garcia-Ramirez, M.

A. Dufresne, J. Y. Cavaille, D. Dupeyre, M. Garcia-Ramirez, and J. Romero, "Morphology, phase continuity and mechanical behavior of polyamide 6/chitosan blends," Polymer. 40, 1657-1666 (1999).
[CrossRef]

Gerbi, A.

A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
[CrossRef]

Gu, X.

Guo, B. H.

J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
[CrossRef]

Gupper, A.

A. Gupper, P. Wilhelm, M. Schmied, and E. Ingolic. "Morphology of a PA/PTFE blend studied by Raman imaging," Macromol. Symp. 184, 275-286 (2002).
[CrossRef]

Ha, N. Y.

N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
[CrossRef] [PubMed]

Hamano, K.

H. Jinnai, H. Kitagishi, K. Hamano, Y. Nishikawa, and Takahashi, "Effect of confinement on phase-separation processes in a polymer blend observed by laser scanning confocal microscopy," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 67, 021801 (2003).
[CrossRef] [PubMed]

Han, S. H.

N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
[CrossRef] [PubMed]

Hashimoto, T.

A. E. Ribbe, M. Hayashi, M. Weber, and T. Hashimoto, "Identification of the morphology of a thin film by fluorescence laser-scanning confocal microscopy," Polymer. 39, 7149-7151(1998).
[CrossRef]

H. Jinnai, Y. Nishikawa, T. Koga, and T. Hashimoto, "Direct Observation of Three-dimensional Bicontinuous Structure Developed via Spinodal Decomposition," Macromolecules 28, 4782-4784 (1995).
[CrossRef]

Hayashi, M.

A. E. Ribbe, M. Hayashi, M. Weber, and T. Hashimoto, "Identification of the morphology of a thin film by fluorescence laser-scanning confocal microscopy," Polymer. 39, 7149-7151(1998).
[CrossRef]

He, G. S.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Heckmann, W.

J. Kressler, N. Higashida, T. Inoue, W. Heckmann, and F. Seitz, "Study of Polymer-Polymer Interfaces: A Comparison of Ellipsometric and TEM Data of PMMA/PS and PMMA/SAN Systems," Macromolecules 26, 2090-2094 (1993).
[CrossRef]

Higashida, N.

J. Kressler, N. Higashida, T. Inoue, W. Heckmann, and F. Seitz, "Study of Polymer-Polymer Interfaces: A Comparison of Ellipsometric and TEM Data of PMMA/PS and PMMA/SAN Systems," Macromolecules 26, 2090-2094 (1993).
[CrossRef]

Hodge, T.

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

Hsiao, W. C.

S. J. Lin, W. C. Hsiao, S. H. Jee, H. S. Yu, T. F. Tsai, J. Y. Lai, and T. H. Young, "Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes," Biomaterials 27, 5079-5088 (2006).
[CrossRef] [PubMed]

Ingolic, E.

A. Gupper, P. Wilhelm, M. Schmied, and E. Ingolic. "Morphology of a PA/PTFE blend studied by Raman imaging," Macromol. Symp. 184, 275-286 (2002).
[CrossRef]

Inoue, T.

J. Kressler, N. Higashida, T. Inoue, W. Heckmann, and F. Seitz, "Study of Polymer-Polymer Interfaces: A Comparison of Ellipsometric and TEM Data of PMMA/PS and PMMA/SAN Systems," Macromolecules 26, 2090-2094 (1993).
[CrossRef]

Iwata, T.

J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
[CrossRef]

Jamison, R.

W. Tan, A. Sendermir-Urkmez, L. J. Fahrmer, R. Jamison, and D. Leckband, "Structual and functional optical imaging of three-dimensional engineered tissue development," Tissue. Eng. 10, 1747-1757 (2004).
[CrossRef]

Jee, S. H.

S. J. Lin, W. C. Hsiao, S. H. Jee, H. S. Yu, T. F. Tsai, J. Y. Lai, and T. H. Young, "Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes," Biomaterials 27, 5079-5088 (2006).
[CrossRef] [PubMed]

Jeon, D. W.

N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
[CrossRef] [PubMed]

Jiang, C. C.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Jinnai, H.

H. Jinnai, H. Kitagishi, K. Hamano, Y. Nishikawa, and Takahashi, "Effect of confinement on phase-separation processes in a polymer blend observed by laser scanning confocal microscopy," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 67, 021801 (2003).
[CrossRef] [PubMed]

H. Jinnai, Y. Nishikawa, T. Koga, and T. Hashimoto, "Direct Observation of Three-dimensional Bicontinuous Structure Developed via Spinodal Decomposition," Macromolecules 28, 4782-4784 (1995).
[CrossRef]

Jung, C. S.

N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
[CrossRef] [PubMed]

Jung, W. G.

A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
[CrossRef] [PubMed]

Kao, B.

A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
[CrossRef] [PubMed]

Karim, A.

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

Keskkula, H.

R. A. Kudva, H. Keskkula, and D. R. Paul, "Compatibilization of nylon 6/ABS blends using glycidyl methacrylate/methyl methacrylate copolymers," Polymer 39, 2447-2460 (1998).
[CrossRef]

Kitagishi, H.

H. Jinnai, H. Kitagishi, K. Hamano, Y. Nishikawa, and Takahashi, "Effect of confinement on phase-separation processes in a polymer blend observed by laser scanning confocal microscopy," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 67, 021801 (2003).
[CrossRef] [PubMed]

Koga, T.

H. Jinnai, Y. Nishikawa, T. Koga, and T. Hashimoto, "Direct Observation of Three-dimensional Bicontinuous Structure Developed via Spinodal Decomposition," Macromolecules 28, 4782-4784 (1995).
[CrossRef]

Krasieva, T. B.

S. Tang, T. B. Krasieva, Z. Chen, B. J. Tromberg, "Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source," J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

Kressler, J.

J. Kressler, N. Higashida, T. Inoue, W. Heckmann, and F. Seitz, "Study of Polymer-Polymer Interfaces: A Comparison of Ellipsometric and TEM Data of PMMA/PS and PMMA/SAN Systems," Macromolecules 26, 2090-2094 (1993).
[CrossRef]

Kudva, R. A.

R. A. Kudva, H. Keskkula, and D. R. Paul, "Compatibilization of nylon 6/ABS blends using glycidyl methacrylate/methyl methacrylate copolymers," Polymer 39, 2447-2460 (1998).
[CrossRef]

Lai, J. Y.

S. J. Lin, W. C. Hsiao, S. H. Jee, H. S. Yu, T. F. Tsai, J. Y. Lai, and T. H. Young, "Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes," Biomaterials 27, 5079-5088 (2006).
[CrossRef] [PubMed]

Leckband, D.

W. Tan, A. Sendermir-Urkmez, L. J. Fahrmer, R. Jamison, and D. Leckband, "Structual and functional optical imaging of three-dimensional engineered tissue development," Tissue. Eng. 10, 1747-1757 (2004).
[CrossRef]

Lee, H. S.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Liang, C.

K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
[CrossRef] [PubMed]

Lin, S. J.

S. J. Lin, W. C. Hsiao, S. H. Jee, H. S. Yu, T. F. Tsai, J. Y. Lai, and T. H. Young, "Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes," Biomaterials 27, 5079-5088 (2006).
[CrossRef] [PubMed]

Lin, T. Y.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Liou, W. S.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Liu, H. C.

J. H. Wang, C. W. Wei, H. C. Liu, and T. H. Young, "Behavior of MG-63 cells on nylon/chitosan-blended membranes," J. Biomed. Mater. Res. A. 64, 606-615 (2003).
[CrossRef] [PubMed]

Lo, W.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Long, R.

I. Constantinidis, C. L. Stabler, R. Long, and A. Sambanis, "Noninvasive monitoring of a retrievable bioartificial pancreas in vivo," Ann. N. Y. Acad. Sci. 961, 298-301 (2002).
[CrossRef] [PubMed]

Ma, H.

J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
[CrossRef]

McCormick, J.

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

Mi, F. L.

F. L. Mi, "Synthesis and characterization of a novel chitosan-gelatin bioconjugate with fluorescence emission," Biomacromolecules 6, 975-987 (2005).
[CrossRef] [PubMed]

Mikes, F.

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

Millard, A. C.

S. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys J. 90, 693-703 (2006).
[CrossRef]

Miller, R. D.

D. M. Burland, R. D. Miller, and C. A. Walsh. "Second-order nonlinearity in poled-polymer systems," Chem. Rev. 94, 31-75 (1994).
[CrossRef]

Mohler, W. A.

S. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys J. 90, 693-703 (2006).
[CrossRef]

Moreland, C.

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

Nelson, J. S.

A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
[CrossRef] [PubMed]

Nishikawa, Y.

H. Jinnai, H. Kitagishi, K. Hamano, Y. Nishikawa, and Takahashi, "Effect of confinement on phase-separation processes in a polymer blend observed by laser scanning confocal microscopy," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 67, 021801 (2003).
[CrossRef] [PubMed]

H. Jinnai, Y. Nishikawa, T. Koga, and T. Hashimoto, "Direct Observation of Three-dimensional Bicontinuous Structure Developed via Spinodal Decomposition," Macromolecules 28, 4782-4784 (1995).
[CrossRef]

Pan, S. J.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Park, B.

N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
[CrossRef] [PubMed]

Paul, D. R.

R. A. Kudva, H. Keskkula, and D. R. Paul, "Compatibilization of nylon 6/ABS blends using glycidyl methacrylate/methyl methacrylate copolymers," Polymer 39, 2447-2460 (1998).
[CrossRef]

Pellistri, F.

A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
[CrossRef]

Pierola, I. F.

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

Plotnikov, S.

S. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys J. 90, 693-703 (2006).
[CrossRef]

Prasad, P. N.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Raghavan, D.

P. Achalla, J. McCormick, T. Hodge, C. Moreland, P. Esnault, A. Karim, and D. Raghavan, "Characterization of Elastomeric Blends by Atomic Force Microscopy," J. Polym. Sci. B 44, 492-503 (2006).

Ramoino, P.

A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
[CrossRef]

Ribbe, A. E.

A. E. Ribbe, M. Hayashi, M. Weber, and T. Hashimoto, "Identification of the morphology of a thin film by fluorescence laser-scanning confocal microscopy," Polymer. 39, 7149-7151(1998).
[CrossRef]

A. E. Ribbe, "Laser scanning confocal microscopy in polymer science," Trends Polym. Sci. 5, 333-337 (1997).

Richards-Kortum, R.

K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
[CrossRef] [PubMed]

Robello, M.

A. Diaspro, F. Pellistri, F. Federici, A. Gerbi, P. Ramoino, and M. Robello, "Evaluation of optical properties and micropatterning capabilities of a TPE microscope based on a compact confocal scanning head," Proc. SPIE 4262, 383-388(2001).
[CrossRef]

Romero, J.

A. Dufresne, J. Y. Cavaille, D. Dupeyre, M. Garcia-Ramirez, and J. Romero, "Morphology, phase continuity and mechanical behavior of polyamide 6/chitosan blends," Polymer. 40, 1657-1666 (1999).
[CrossRef]

Samarabandu, J. K.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Sambanis, A.

I. Constantinidis, C. L. Stabler, R. Long, and A. Sambanis, "Noninvasive monitoring of a retrievable bioartificial pancreas in vivo," Ann. N. Y. Acad. Sci. 961, 298-301 (2002).
[CrossRef] [PubMed]

Schmied, M.

A. Gupper, P. Wilhelm, M. Schmied, and E. Ingolic. "Morphology of a PA/PTFE blend studied by Raman imaging," Macromol. Symp. 184, 275-286 (2002).
[CrossRef]

Seitz, F.

J. Kressler, N. Higashida, T. Inoue, W. Heckmann, and F. Seitz, "Study of Polymer-Polymer Interfaces: A Comparison of Ellipsometric and TEM Data of PMMA/PS and PMMA/SAN Systems," Macromolecules 26, 2090-2094 (1993).
[CrossRef]

Sendermir-Urkmez, A.

W. Tan, A. Sendermir-Urkmez, L. J. Fahrmer, R. Jamison, and D. Leckband, "Structual and functional optical imaging of three-dimensional engineered tissue development," Tissue. Eng. 10, 1747-1757 (2004).
[CrossRef]

Serrano, B.

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

Sese, L.

B. Serrano, J. Baselga, J. Bravo, F. Mikes, L. Sese, I. Esteban, and I. F. Pierola, "Chemical Imaging of Phase-Separated Polymer Blends by Fluorescence Microscopy," J. Fluoresc. 10, 135 (2000).
[CrossRef]

Stabler, C. L.

I. Constantinidis, C. L. Stabler, R. Long, and A. Sambanis, "Noninvasive monitoring of a retrievable bioartificial pancreas in vivo," Ann. N. Y. Acad. Sci. 961, 298-301 (2002).
[CrossRef] [PubMed]

Stranick, S. J.

Sung, K. B.

K. B. Sung, C. Liang, M. Descour, T. Collier, M. Follen, and R. Richards-Kortum, "Fiber-optic confocal reflectance microscope with miniature objective for in vivo imaging of human tissues," IEEE Trans. Biomed. Eng. 49, 1168-1172 (2002).
[CrossRef] [PubMed]

Swiatkiewicz, J.

J. D. Bhawalkar, J. Swiatkiewicz, S. J. Pan, J. K. Samarabandu, W. S. Liou, G. S. He, R. Berezney, P. C. Cheng, and P. N. Prasad, "Three-Dimensional Laser Scanning Two-Photon Fluorescence Confocal Microscopy of Polymer Materials using a new, efficient upconverting Fluorophore," Scanning 18, 562-566 (1996).
[CrossRef] [PubMed]

Takezoe, H.

N. Y. Ha, S. H. Han, D. W. Jeon, C. S. Jung, B. Park, H. Takezoe, and J. W. Wu, "Analysis of surface second-harmonic generation by orientational distribution function in a chiral polymer film," Phys. Rev. E. Stat. Nonlin. Soft. Matter. Phys. 72, 036601 (2005).
[CrossRef] [PubMed]

Tan, W.

W. Tan, A. Sendermir-Urkmez, L. J. Fahrmer, R. Jamison, and D. Leckband, "Structual and functional optical imaging of three-dimensional engineered tissue development," Tissue. Eng. 10, 1747-1757 (2004).
[CrossRef]

Tang, S.

S. Tang, T. B. Krasieva, Z. Chen, B. J. Tromberg, "Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source," J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

Teng, S. W.

H. S. Lee, S. W. Teng, H. C. Chen, W. Lo, T. Y. Lin, L. L. Chiou, C. C. Jiang, and C. Y. Dong. "Imaging human bone marrow stem cell morphogenesis in PGA scaffold by multiphoton microscopy," Tissue. Eng. 12, 2835-2841 (2006).
[CrossRef]

Tromberg, B. J.

S. Tang, T. B. Krasieva, Z. Chen, B. J. Tromberg, "Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source," J. Biomed. Opt. 11, 020502 (2006).
[CrossRef] [PubMed]

A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
[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," P. Natl. Acad. Sci. U.S.A. 99, 11014-11019 (2002).
[CrossRef]

Tsai, T. F.

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A. T. Yeh, B. Kao, W. G. Jung, Z. Chen, J. S. Nelson, and B. J. Tromberg. "Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model," J. Biomed. Opt. 9, 248-253 (2004).
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S. J. Lin, W. C. Hsiao, S. H. Jee, H. S. Yu, T. F. Tsai, J. Y. Lai, and T. H. Young, "Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes," Biomaterials 27, 5079-5088 (2006).
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J. Xu, J. Bao, B. H. Guo, H. Ma, T. L. Yun, L. Gao, G. Q. Chen, T. Iwata. "Imaging of nonlinear optical response in biopolyesters via second harmonic generation microscopy and its dependence on the crystalline structures," Polymer. 48, 348-355 (2007).
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S. Tang, T. B. Krasieva, Z. Chen, B. J. Tromberg, "Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source," J. Biomed. Opt. 11, 020502 (2006).
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A. Gupper, P. Wilhelm, M. Schmied, and E. Ingolic. "Morphology of a PA/PTFE blend studied by Raman imaging," Macromol. Symp. 184, 275-286 (2002).
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Figures (7)

Fig. 1.
Fig. 1.

Instrumentation setup of the combined multiphoton (MP) and reflected confocal (RC) microscope.

Fig. 2.
Fig. 2.

Scanning electron micrographs of top surfaces of pure nylon (A), pure chitosan (E), and blended membrane composed of different nylon/chitosan weight ratios ((B): N75C25; (C): N50C50; (D): N25C75). The bottom three images were taken from blended membrane with etching treatment: (F) N75C25 membrane treated with chitinase; (G) N50C50 treated with n-cresol; (H) N25C75 membrane treated with n-cresol. N and C are used to designate nylon and chitosan respectively (Magnification X 1,000).

Fig. 3.
Fig. 3.

Combined multiphoton (MP) and reflected confocal (RC) imaging of pure nylon membrane in two-dimensions (A) and three-dimensions (B). Spherical domains can be identified in reflected confocal (RC), autofluorescence (AF), and SHG images. The weaker SHG signal prevents a sharp definition of the domain boundaries.

Fig. 4.
Fig. 4.

Combined MP and RC imaging of pure chitosan membrane in two-dimensions (A) and three-dimensions (B). Relatively uniform AF and SHG signals are observed within full-thickness of the membrane while the RC signal is most intense at the interfaces.

Fig. 5.
Fig. 5.

Two-dimensional XY-sectional (A), three-dimensional (B) and cross-sectional (C) MP and RC images of the blended N75C25 membrane. Particulate aggregates (chitosan domains) composed of intense AF and SHG signals can be identified at the surfaces (white arrows) and are coated on large spherical domain (nylon domain –yellow dashed region) composed of less AF and minimal SHG signals (white dashed circle). In both three-dimensional and cross-sectional view, chitosan domain composed of highly intense AF and SHG signals were shown to be distributed preferentially at the membrane surface.

Fig. 6.
Fig. 6.

Two-dimensional (X-Y plane) MP and RC images of blended N75C25 membrane with etching treatment for chitosan removal. The presence of hollow regions (white dashed circle) may be due to etching of the autofluorescent and second harmonic generating chitosan domains. The remaining autofluorescent substrates are likely the results of the incomplete dissolution of chitosan during the etching process.

Fig. 7.
Fig. 7.

Combined MP and RC imaging of blended N50C50 membrane in two- (A) and three- (B) dimensions. Compared with that of the N75C25 blend, the surface topography is relatively smooth. Spherical domains composed of weaker AF and SHG signals (nylon domains), are embedded within the matrix of stronger AF and SHG signals (most likely the chitosan domains).

Tables (1)

Tables Icon

Table 1. Comparison of the average intensity of AF and SHG signals of pure nylon and chitosan membranes.

Metrics