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Journal of Biomedical Optics • Open Access

Simultaneous determination of the second-harmonic generation emission directionality and reduced scattering coefficient from three-dimensional imaging of thick tissues

Paper Abstract

Second-harmonic generation (SHG) microscopy has intrinsic contrast for imaging fibrillar collagen and has shown great promise for disease characterization and diagnostics. In addition to morphology, additional information is achievable as the initially emitted SHG radiation directionality is related to subresolution fibril size and distribution. We show that by two parameter fittings, both the emission pattern (F SHG /B SHG ) creation and the reduced scattering coefficient μ′s , can be obtained from the best fits between three-dimensional experimental data and Monte Carlo simulations. The improved simulation framework accounts for collection apertures for the detected forward and backward components. We apply the new simulation framework to mouse tail tendon for validation and show that the spectral slope of μ′s obtained is similar to that from bulk optical measurements and that the (F SHG /B SHG ) creation values are also similar to previous results. Additionally, we find that the SHG emission becomes increasingly forward directed at longer wavelengths, which is consistent with decreased dispersion in refractive index between the laser and SHG wavelengths. As both the spectral slope of μ ′ s and (F SHG /B SHG ) creation have been linked to the underlying tissue structure, simultaneously obtaining these parameters on a microscope platform from the same tissue provides a powerful method for tissue characterization.

Paper Details

Date Published: 12 November 2013
PDF: 10 pages
J. Biomed. Opt. 18(11) 116008 doi: 10.1117/1.JBO.18.11.116008
Published in: Journal of Biomedical Optics Volume 18, Issue 11
Show Author Affiliations
Gunnsteinn Hall, Johns Hopkins Univ. (United States)
Kevin W. Eliceiri, Univ. of Wisconsin-Madison (United States)
Paul J. Campagnola, Univ. of Wisconsin-Madison (United States)

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