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Determination of the source of SHG verniers in zebrafish skeletal muscle

SHG microscopy is an emerging microscopic technique for medically relevant imaging because certain endogenous proteins, such as muscle myosin lattices within muscle cells, are sufficiently spatially ordered to generate detectable SHG without the use of any fluorescent dye. Given that SHG signal is s...

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Detalles Bibliográficos
Autores principales: Dempsey, William P., Hodas, Nathan O., Ponti, Aaron, Pantazis, Periklis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4676038/
https://www.ncbi.nlm.nih.gov/pubmed/26657568
http://dx.doi.org/10.1038/srep18119
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author Dempsey, William P.
Hodas, Nathan O.
Ponti, Aaron
Pantazis, Periklis
author_facet Dempsey, William P.
Hodas, Nathan O.
Ponti, Aaron
Pantazis, Periklis
author_sort Dempsey, William P.
collection PubMed
description SHG microscopy is an emerging microscopic technique for medically relevant imaging because certain endogenous proteins, such as muscle myosin lattices within muscle cells, are sufficiently spatially ordered to generate detectable SHG without the use of any fluorescent dye. Given that SHG signal is sensitive to the structural state of muscle sarcomeres, SHG functional imaging can give insight into the integrity of muscle cells in vivo. Here, we report a thorough theoretical and experimental characterization of myosin-derived SHG intensity profiles within intact zebrafish skeletal muscle. We determined that “SHG vernier” patterns, regions of bifurcated SHG intensity, are illusory when sarcomeres are staggered with respect to one another. These optical artifacts arise due to the phase coherence of SHG signal generation and the Guoy phase shift of the laser at the focus. In contrast, two-photon excited fluorescence images obtained from fluorescently labeled sarcomeric components do not contain such illusory structures, regardless of the orientation of adjacent myofibers. Based on our results, we assert that complex optical artifacts such as SHG verniers should be taken into account when applying functional SHG imaging as a diagnostic readout for pathological muscle conditions.
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spelling pubmed-46760382015-12-16 Determination of the source of SHG verniers in zebrafish skeletal muscle Dempsey, William P. Hodas, Nathan O. Ponti, Aaron Pantazis, Periklis Sci Rep Article SHG microscopy is an emerging microscopic technique for medically relevant imaging because certain endogenous proteins, such as muscle myosin lattices within muscle cells, are sufficiently spatially ordered to generate detectable SHG without the use of any fluorescent dye. Given that SHG signal is sensitive to the structural state of muscle sarcomeres, SHG functional imaging can give insight into the integrity of muscle cells in vivo. Here, we report a thorough theoretical and experimental characterization of myosin-derived SHG intensity profiles within intact zebrafish skeletal muscle. We determined that “SHG vernier” patterns, regions of bifurcated SHG intensity, are illusory when sarcomeres are staggered with respect to one another. These optical artifacts arise due to the phase coherence of SHG signal generation and the Guoy phase shift of the laser at the focus. In contrast, two-photon excited fluorescence images obtained from fluorescently labeled sarcomeric components do not contain such illusory structures, regardless of the orientation of adjacent myofibers. Based on our results, we assert that complex optical artifacts such as SHG verniers should be taken into account when applying functional SHG imaging as a diagnostic readout for pathological muscle conditions. Nature Publishing Group 2015-12-11 /pmc/articles/PMC4676038/ /pubmed/26657568 http://dx.doi.org/10.1038/srep18119 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Dempsey, William P.
Hodas, Nathan O.
Ponti, Aaron
Pantazis, Periklis
Determination of the source of SHG verniers in zebrafish skeletal muscle
title Determination of the source of SHG verniers in zebrafish skeletal muscle
title_full Determination of the source of SHG verniers in zebrafish skeletal muscle
title_fullStr Determination of the source of SHG verniers in zebrafish skeletal muscle
title_full_unstemmed Determination of the source of SHG verniers in zebrafish skeletal muscle
title_short Determination of the source of SHG verniers in zebrafish skeletal muscle
title_sort determination of the source of shg verniers in zebrafish skeletal muscle
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4676038/
https://www.ncbi.nlm.nih.gov/pubmed/26657568
http://dx.doi.org/10.1038/srep18119
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