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      Molecular component distribution imaging of living cells by multivariate curve resolution analysis of space-resolved Raman spectra.

      1 ,
      Journal of biomedical optics
      SPIE-Intl Soc Optical Eng

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          Abstract

          Label-free Raman microspectroscopy combined with a multivariate curve resolution (MCR) analysis can be a powerful tool for studying a wide range of biomedical molecular systems. The MCR with the alternating least squares (MCR-ALS) technique, which retrieves the pure component spectra from complicatedly overlapped spectra, has been successfully applied to in vivo and molecular-level analysis of living cells. The principles of the MCR-ALS analysis are reviewed with a model system of titanium oxide crystal polymorphs, followed by two examples of in vivo Raman imaging studies of living yeast cells, fission yeast, and budding yeast. Due to the non-negative matrix factorization algorithm used in the MCR-ALS analysis, the spectral information derived from this technique is just ready for physical and/or chemical interpretations. The corresponding concentration profiles provide the molecular component distribution images (MCDIs) that are vitally important for elucidating life at the molecular level, as stated by Schroedinger in his famous book, "What is life?" Without any a priori knowledge about spectral profiles, time- and space-resolved Raman measurements of a dividing fission yeast cell with the MCR-ALS elucidate the dynamic changes of major cellular components (lipids, proteins, and polysaccharides) during the cell cycle. The MCR-ALS technique also resolves broadly overlapped OH stretch Raman bands of water, clearly indicating the existence of organelle-specific water structures in a living budding yeast cell.

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          Author and article information

          Journal
          J Biomed Opt
          Journal of biomedical optics
          SPIE-Intl Soc Optical Eng
          1560-2281
          1083-3668
          Jan 2014
          : 19
          : 1
          Affiliations
          [1 ] Waseda University, Consolidated Research Institute for Advanced Science and Medical Care, 513 Wasedatsurumaki-cho, Shinjuku-ku, Tokyo 162-0041, Japan.
          Article
          1752549
          10.1117/1.JBO.19.1.011016
          24108582
          052db174-7ba7-486b-9bc8-a0beecc67e34

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