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Coherent Raman Imaging of Live Muscle Sarcomeres Assisted by SFG Microscopy

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Title
Coherent Raman Imaging of Live Muscle Sarcomeres Assisted by SFG Microscopy
Author(s)
Hyunmin Kim; Do-Young Kim; Kyung-Il Joo; Jung-Hye Kim; Soon Moon Jeong; Eun Seong Lee; Jeong-Hoon Hahm; Kyuhyung Kim; Dae Woon Moon
Publication Date
2017-12
Journal
SCIENTIFIC REPORTS, v.7, no.1, pp.9211 -
Publisher
NATURE PUBLISHING GROUP
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 alpha-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 similar to 90 degrees 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-Kronig 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. © The Author(s) 2017
URI
http://pr.ibs.re.kr/handle/8788114/3838
DOI
10.1038/s41598-017-09571-w
ISSN
2045-2322
Appears in Collections:
Center for Plant Aging Research (식물 노화·수명 연구단) > Journal Papers (저널논문)
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