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Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN OPTICS

  • Vol. 42, Iss. 25 — Sep. 1, 2003
  • pp: 5209–5219

Quantitative second-harmonic generation microscopy in collagen

Patrick Stoller, Peter M. Celliers, Karen M. Reiser, and Alexander M. Rubenchik  »View Author Affiliations


Applied Optics, Vol. 42, Issue 25, pp. 5209-5219 (2003)
http://dx.doi.org/10.1364/AO.42.005209


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Abstract

The second-harmonic signal in collagen, even in highly organized samples such as rat tail tendon fascicles, varies significantly with position. Previous studies suggest that this variability may be due to the parallel and antiparallel orientation of neighboring collagen fibrils. We applied high-resolution second-harmonic generation microscopy to confirm this hypothesis. Studies in which the focal spot diameter was varied from ∼1 to ∼6 μm strongly suggest that regions in which collagen fibrils have the same orientation in rat tail tendon are likely to be less than ∼1 μm in diameter. These measurements required accurate determination of the focal spot size achieved by use of different microscope objectives; we developed a technique that uses second-harmonic generation in a quartz reference to measure the focal spot diameter directly. We also used the quartz reference to determine a lower limit (d XXX > 0.4 pm/V) for the magnitude of the second-order nonlinear susceptibility in collagen.

© 2003 Optical Society of America

OCIS Codes
(170.0180) Medical optics and biotechnology : Microscopy
(170.5810) Medical optics and biotechnology : Scanning microscopy
(170.7160) Medical optics and biotechnology : Ultrafast technology
(180.5810) Microscopy : Scanning microscopy
(190.1900) Nonlinear optics : Diagnostic applications of nonlinear optics
(190.4710) Nonlinear optics : Optical nonlinearities in organic materials

History
Original Manuscript: September 10, 2002
Revised Manuscript: January 6, 2003
Published: September 1, 2003

Citation
Patrick Stoller, Peter M. Celliers, Karen M. Reiser, and Alexander M. Rubenchik, "Quantitative second-harmonic generation microscopy in collagen," Appl. Opt. 42, 5209-5219 (2003)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-42-25-5209

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