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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
Author(s)
Kim, HyunminKim, Do-YoungJoo, Kyung-IlKim, Jung-HyeJeong, Soon MoonLee, Eun SeongHahm, Jeong-HoonKim, KyuhyungMoon, Dae Woon
Issued Date
2017-08
Citation
Scientific Reports, v.7, no.1
Type
Article
Keywords
2ND-HARMONIC GENERATIONCAENORHABDITIS-ELEGANSSCATTERINGSPECTROSCOPYPROTEINSCOLLAGENBINDINGMODELBAND
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
Related Researcher
  • 김현민 Kim, Hyunmin
  • Research Interests Nonlinear optics; Femtosecond ultrafast carrier dynamics; Super resolution microscopy; Optical soliton transport; Biological imaging
Files in This Item:
10.1038_s41598_017_09571_w.pdf

10.1038_s41598_017_09571_w.pdf

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Appears in Collections:
Division of Energy & Environmental Technology Soft Optoelectronic Materials Lab. 1. Journal Articles
Department of New Biology NanoBio Imaging Laboratory 1. Journal Articles
Department of Brain Sciences The K. Kim Lab of Neurobehavior and Neural Circuits 1. Journal Articles
Division of Biomedical Technology 1. Journal Articles

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