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Biophys J, June 2002, p. 3330-3342, Vol. 82, No. 6

*Medical Technology Program, Lawrence Livermore National
Laboratory, Livermore, California 94551, and
Department of Neurological Surgery, School of
Medicine, University of California, Davis, California 95817 USA
Collagen possesses a strong second-order nonlinear
susceptibility, a nonlinear optical property characterized by second
harmonic generation in the presence of intense laser beams. We present a new technique involving polarization modulation of an ultra-short pulse laser beam that can simultaneously determine collagen fiber orientation and a parameter related to the second-order nonlinear susceptibility. We demonstrate the ability to discriminate among different patterns of fibrillar orientation, as exemplified by tendon,
fascia, cornea, and successive lamellar rings in an intervertebral disc. Fiber orientation can be measured as a function of depth with an
axial resolution of ~10 µm. The parameter related to the second-order nonlinear susceptibility is sensitive to fiber
disorganization, oblique incidence of the beam on the sample, and
birefringence of the tissue. This parameter represents an aggregate
measure of tissue optical properties that could potentially be used for optical imaging in vivo.
Biophys J, June 2002, p. 3330-3342, Vol. 82, No. 6
© 2002 by the Biophysical Society 0006-3495/02/06/3330/13 $2.00
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