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Coherent raman imaging of live muscle sarcomeres assisted by SFG microscopy

Title
Coherent raman imaging of live muscle sarcomeres assisted by SFG microscopy
Authors
Kim, HyunminKim, Do-YoungJoo, Kyung-IlKim, Jung-HyeJeong, Soon MoonLee, Eun SeongHahm, Jeong-HoonKim, KyuhyungMoon, Dae Woon
DGIST Authors
Kim, Hyunmin; Kim, Do-Young; Kim, Jung-Hye; Jeong, Soon Moon; Kim, Kyuhyung; Moon, Dae Woon
Issue Date
2017
Citation
Scientific Reports, 7(1)
Type
Article
Article Type
Article
ISSN
2045-2322
Abstract
In this study, we used spectrally focused coherent anti-Stokes Raman scattering (spCARS) microscopy assisted by sum-frequency generation (SFG) to monitor the variations in the structural morphology and molecular vibrations of a live muscle of Caenorhabditis elegans. The subunits of the muscle sarcomeres, such as the M-line, myosin, dense body, and α-actinin, were alternatively observed using spCARS microscopy for different sample orientations, with the guidance of a myosin positional marker captured by SFG microscopy. Interestingly enough, the beam polarization dependence of the spCARS contrasts for two parallel subunits (dense body and myosin) showed a ~90° phase difference. The chemically sensitive spCARS spectra induced by the time-varying overlap of two pulses allowed (after a robust subtraction of the non-resonant background using a modified Kramers-Krönig transformation method) high-fidelity detection of various genetically modified muscle sarcomeres tuned to the C-H vibration (2800-3100 cm-1). Conversely, SFG image mapping assisted by phase-retrieved spCARS spectra also facilitated label-free monitoring of the changes in the muscle content of C. elegans that are associated with aging, based on the hypothesis that the C-H vibrational modes could serve as qualitative chemical markers sensitive to the amount and/or structural modulation of the muscle. © 2017 The Author(s).
URI
http://hdl.handle.net/20.500.11750/4438
DOI
10.1038/s41598-017-09571-w
Publisher
Nature Publishing Group
Files:
There are no files associated with this item.
Collection:
Division of Nano∙Energy Convergence Research1. Journal Articles
Brain and Cognitive SciencesThe K. Kim Lab of Neurobehavior and Neural Circuits1. Journal Articles
New BiologyNanoBio Imaging Laboratory1. Journal Articles


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