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Journal of Biomedical Optics

Polarization gating enables sarcomere length measurements by laser diffraction in fibrotic muscle
Author(s): Kevin W. Young; Sudarshan Dayanidhi; Richard L. Lieber
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Paper Abstract

Sarcomere length is a key parameter commonly measured in muscle physiology since it dictates striated muscle active force. Laser diffraction (LD)–based measurements of sarcomere length are time-efficient and sample a greater number of sarcomeres compared with traditional microscopy–based techniques. However, a limitation to LD techniques is that signal quality is severely degraded by scattering events as photons propagate through tissue. Consequently, sarcomere length measurements are unattainable when the number of scattering events is sufficiently large in muscle tissue with a high scattering probability. This occurs in fibrotic skeletal muscle seen in muscular dystrophies and secondary to tissue trauma, thus eliminating the use of LD to study these skeletal muscle ailments. Here, we utilize polarization gating to extract diffracted signals that are buried in noise created by scattering. Importantly, we demonstrate that polarization-gated laser diffraction (PGLD) enables sarcomere length measurements in muscles from chronically immobilized mice hind limbs; these muscles have a substantial increase of intramuscular connective tissue that scatter light and disable sarcomere length measurements by traditional LD. Further, we compare PGLD sarcomere lengths to those measured by bright field (BF) and confocal microscopy as positive controls and reveal a significant bias of BF but not of confocal microscopy.

Paper Details

Date Published: 25 November 2014
PDF: 5 pages
J. Biomed. Opt. 19(11) 117009 doi: 10.1117/1.JBO.19.11.117009
Published in: Journal of Biomedical Optics Volume 19, Issue 11
Show Author Affiliations
Kevin W. Young, Univ. of California, San Diego (United States)
VA San Diego Healthcare System (United States)
Sudarshan Dayanidhi, VA San Diego Healthcare System (United States)
Univ. of California, San Diego (United States)
Richard L. Lieber, Univ. of California, San Diego (United States)
VA San Diego Healthcare System (United States)

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