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Opt Express. 2008 Sep 29;16(20):16151-65.

Collagen and myosin characterization by orientation field second harmonic microscopy.

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  • 1Institut of Physics of Rennes IPR/UMR CNRS 6251, University of Rennes I, Campus deBeaulieu, Bat 11A, 35042 Rennes Cedex, France. christophe.odin@univ-rennes1.fr


Collagen and myosin fibrils are endogenous harmonophores that both give rise to Second Harmonic Generation (SHG). By combining four polarization SHG images provided by a scanning microscope, we show that the orientation of the principal axis of the nonlinear susceptibility tensor chi(2) can be determined for each pixel of the image. The ratio rho = chi33/chi15 of the principal components of chi(2) of collagen and myosin was obtained with the same method, and found within the range 1.6-1.8 and 0.5-0.6 respectively. The orientation of the principal axis of chi(2) is shown to be correlated to the orientation of the fibrils themselves. This provides a straightforward method, which we call Orientation Field-Second Harmonic Microscopy (OF-SHM), to reconstruct orientation fields of fibrils at various scales and resolutions in different biological systems (from muscle sarcomere to the whole embryo).

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