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Journal of Biomedical Optics • Open Access

In vivo, pixel-resolution mapping of thick filaments' orientation in nonfibrilar muscle using polarization-sensitive second harmonic generation microscopy
Author(s): Sotiris Psilodimitrakopoulos; Susana I. C. O. Santos; Ivan Amat-Roldán; Anisha Thayil Karunakaran Nair; David Artigas-García; Pablo Loza-Alvarez

Paper Abstract

The polarization dependence of second harmonic generation (SHG) microscopy is used to uncover structural information in different muscle cells in a living Caenorhabditis elegans (C. elegans) nematode. This is done by using a generalized biophysical model in which element ratios for the associated second-order nonlinear tensor and angular orientations for thick filaments are retrieved using a pixel-by-pixel fitting algorithm. As a result, multiple arbitrary orientations of thick filaments, at the pixel-resolution level, are revealed in the same image. The validity of our method is first corroborated in well-organized thick filaments such as the nonfibrilar body wall muscles. Next, a region of the nonstriated muscular cells of the pharynx is analyzed by showing different regions with homogenous orientations of thick filament as well as their radial distribution. As a result, different sets of the nonstriated muscle cell groups in the pharynx of this nematode were exposed. This methodology is presented as a filtering mechanism to uncover biological information unreachable by common intensity SHG microscopy. Finally, a method to experimentally retrieve the distribution of the effective orientation of active SHG molecules is proposed and tested.

Paper Details

Date Published: 1 January 2009
PDF: 11 pages
J. Biomed. Opt. 14(1) 014001 doi: 10.1117/1.3059627
Published in: Journal of Biomedical Optics Volume 14, Issue 1
Show Author Affiliations
Sotiris Psilodimitrakopoulos, ICFO - Instituto de Ciencias Fotónicas (Spain)
Susana I. C. O. Santos, ICFO - Instituto de Ciencias Fotónicas (Spain)
Ivan Amat-Roldán, ICFO - Instituto de Ciencias Fotónicas (Spain)
Anisha Thayil Karunakaran Nair, ICFO - Instituto de Ciencias Fotónicas (Spain)
David Artigas-García, Univ. Politècnica de Catalunya (Spain)
Pablo Loza-Alvarez, ICFO - Instituto de Ciencias Fotónicas (Spain)


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