1 January 2009 In vivo, pixel-resolution mapping of thick filaments' orientation in nonfibrilar muscle using polarization-sensitive second harmonic generation microscopy
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J. of Biomedical Optics, 14(1), 014001 (2009). doi:10.1117/1.3059627
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.
Sotiris Psilodimitrakopoulos, Susana I. C. O. Santos, Ivan Amat-Roldán, Anisha Thayil Karunakaran Nair, David Artigas-García, Pablo Loza-Alvarez, "In vivo, pixel-resolution mapping of thick filaments' orientation in nonfibrilar muscle using polarization-sensitive second harmonic generation microscopy," Journal of Biomedical Optics 14(1), 014001 (1 January 2009). http://dx.doi.org/10.1117/1.3059627
Submission: Received ; Accepted
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KEYWORDS
Second-harmonic generation

Polarization

Signal detection

In vivo imaging

Molecules

Microscopy

Harmonic generation

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