Organelle specific simultaneous Raman/green fluorescence protein microspectroscopy for living cell physicochemical studies

Nungnit Wattanavichean, Ikuhisa Nishida, Masahiro Ando, Makoto Kawamukai, Tatsuyuki Yamamoto, Hiro o. Hamaguchi*

*この研究の対応する著者

研究成果: Letter査読

5 被引用数 (Scopus)

抄録

We demonstrate a novel bio-spectroscopic technique, “simultaneous Raman/GFP microspectroscopy”. It enables organelle specific Raman microspectroscopy of living cells. Fission yeast, Schizosaccharomyces pombe, whose mitochondria are green fluorescence protein (GFP) labeled, is used as a test model system. Raman excitation laser and GFP excitation light irradiate the sample yeast cells simultaneously. GFP signal is monitored in the anti-Stokes region where interference from Raman scattering is negligibly small. Of note, 13 568 Raman spectra measured from different points of 19 living yeast cells are categorized according to their GFP fluorescence intensities, with the use of a two-component multivariate curve resolution with alternate least squares (MCR-ALS) analysis in the anti-Stokes region. This categorization allows us to know whether or not Raman spectra are taken from mitochondria. Raman spectra specific to mitochondria are obtained by an MCR-ALS analysis in the Stokes region of 1389 strongly GFP positive spectra. Two mitochondria specific Raman spectra have been obtained. The first one is dominated by protein Raman bands and the second by lipid Raman bands, being consistent with the known molecular composition of mitochondria. In addition, the second spectrum shows a strong band of ergosterol at 1602 cm−1, previously reported as “Raman spectroscopic signature of life of yeast.”.

本文言語English
論文番号e201960163
ジャーナルJournal of Biophotonics
13
4
DOI
出版ステータスPublished - 2020 4月 1

ASJC Scopus subject areas

  • 化学 (全般)
  • 材料科学(全般)
  • 生化学、遺伝学、分子生物学(全般)
  • 工学(全般)
  • 物理学および天文学(全般)

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