[15a-C31-1~9] 4.2 Bio-and Medical Photonics

▲ [15a-C31-4] Hybrid Fluorescence-Raman Microscopy for the Visualisation of Protein-Metabolome Interaction in Living Cells

〇(PC)Liangda Chiu^1,2, Taro Ichimura³, Hideaki Fujita^3,4, Takeaki Ozawa¹, Katsumasa Fujita² (1.The Univ. of Tokyo, 2.Osaka Univ., 3.RIKEN, 4.Osaka Univ. IFReC)

Keywords:Raman spectroscopy, Anti-Stokes fluorescence

In recent years, metabolomics has risen as a hot topic of study. Deciphering how the metabolome is regulated by the central dogma is of huge interest to the community now. For the better visualisation of how the metabolome in a living system interacts with the central dogma, we developed a hybrid fluorescence-Raman imaging system for the real-time metabolome analysis according to protein expression patterns. This is the first imaging system that was able to combine the imaging of genetically encoded fluophores and the comprehensive chemical analysis by spontaneous Raman imaging. Anti-Stokes fluorescence excitation was used to spectrally separate the two signals while keeping them at similar intensities for further analysis. We applied the hybrid microscope to study the chemical state of organelles in intact living cells and how the metabolic state of embryo stem cells change according to Oct4 expression. This system is expected to overcome the technical limit that has prevented real-time metabolome analysis in living cells and pave new ways for the research of such kind.