Collagen and myosin characterization by orientation field second harmonic microscopy

Opt Express. 2008 Sep 29;16(20):16151-65. doi: 10.1364/oe.16.016151.

Abstract

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Chickens
  • Collagen / chemistry*
  • Extracellular Matrix / metabolism
  • Image Interpretation, Computer-Assisted / methods
  • Microscopy / methods*
  • Microscopy, Confocal / instrumentation*
  • Microscopy, Confocal / methods
  • Microscopy, Polarization / instrumentation*
  • Microscopy, Polarization / methods
  • Myosins / chemistry*
  • Principal Component Analysis
  • Sarcomeres / metabolism
  • Sheep
  • Tendons / pathology

Substances

  • Collagen
  • Myosins