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Coherent raman imaging of live muscle sarcomeres assisted by SFG microscopy

Coherent raman imaging of live muscle sarcomeres assisted by SFG microscopy
Kim, HyunminKim, Do-YoungJoo, Kyung-IlKim, Jung-HyeJeong, Soon MoonLee, Eun SeongHahm, Jeong-HoonKim, KyuhyungMoon, Dae Woon
DGIST Authors
Kim, HyunminJeong, Soon MoonKim, KyuhyungMoon, Dae Woon
Issue Date
Scientific Reports, 7(1)
Article Type
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).
Nature Publishing Group
Related Researcher
  • Author Kim, Kyuhyung The K. Kim Lab of Neurobehavior and Neural Circuits
  • Research Interests Neurobehavior; Neural Circuit; Neurodevelopment; 신경회로; 신경행동; 신경발생; 신경유전학
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