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Spatial orientation mapping of fibers using polarization-sensitive second harmonic generation microscopy

  1. Vladimir A. Hovhannisyan1,
  2. Po-Shen Hu1,
  3. Hsing-Yuan Tan2,
  4. Shean-Jen Chen3,*,
  5. Chen-Yuan Dong1,4,*

Article first published online: 13 FEB 2012

DOI: 10.1002/jbio.201100123

Additional Information

How to Cite

Hovhannisyan, V. A., Hu, P.-S., Tan, H.-Y., Chen, S.-J. and Dong, C.-Y. (2012), Spatial orientation mapping of fibers using polarization-sensitive second harmonic generation microscopy. J. Biophoton.. doi: 10.1002/jbio.201100123

Author Information

  1. 1

    Department of Physics, National Taiwan University, Taipei 106, Taiwan

  2. 2

    Department of Ophthalmology, Chang-Gung Memorial Hospital, Taoyuan 333, Taiwan

  3. 3

    Department of Engineering Science, National Cheng-Kung University, Tainan 701, Taiwan

  4. 4

    Center for Quantum Science and Engineering, National Taiwan University, Taipei 106, Taiwan

*Department of Engineering Science, National Cheng-Kung University, Tainan 701, Taiwan

Publication History

  1. Article first published online: 13 FEB 2012
  2. Manuscript Accepted: 17 JAN 2012
  3. Manuscript Revised: 5 JAN 2012
  4. Manuscript Received: 10 NOV 2011

Funded by

  • National Science Council, Taiwan (NSC–98–2112-M-002-008-MY3, NSC 99-2221-E-002–096-MY3)
  • National Taiwan University (NTU-99R70409)
  • Center for Quantum Science and Engineering (CQSE-99R80870)
  • National Health Research Institutes (NHRI-EX100-10041EI)

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Keywords:

  • polarization-resolved microscopy;
  • second harmonic generation;
  • three-dimensional image processing;
  • collagen;
  • fiber orientation

Abstract

In this work, we present a non-invasive approach to determine azimuth and elevation angles of collagen fibers capable of generating second harmonic signal. The azimuth angle was determined using the minimum of second harmonic generation (SHG) signal while rotating the plane of polarization of excitation light. The elevation angle was estimated from the ratio of the minimal SHG intensity to the intensity when laser polarization and fiber directions were parallel to each other using experimentally determined calibration curve. Pixel-resolution images of collagen fiber spatial orientation in tendon from bovine leg, chicken leg, and chicken skin were acquired using our approach of SHG polarization-resolved microscopy. (© 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

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