Disentangling Dynamic Changes of Multiple Cellular Components during the Yeast Cell Cycle by <i>in Vivo</i> Multivariate Raman Imaging

Huang, Chuan Keng; Ando, Masahiro; Hamaguchi, Hiro O.; Shigeto, Shinsuke

doi:10.1021/ac300834f

Cited by 81 publications

(110 citation statements)

References 47 publications

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“…Raman spectroscopy can detect the constituents of objects and has been used to detect abnormalities in biological tissue and cells. [23][24][25][26][27] In the current study, Raman mapping data of peripheral nerve tissue in longitudinal sections of intact sciatic nerve tissue in the range 2800-3000 cm −1 showed three strong peaks at 2853, 2885 and 2940 cm −1 [ Fig. 1(b)].…”

Section: Discussionmentioning

confidence: 56%

Application of Raman spectroscopy for visualizing biochemical changes during peripheral nerve injuryin vitroandin vivo

et al. 2013

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Abstract. Raman spectroscopy can be used for analysis of objects by detecting the vibrational spectrum using labelfree methods. This imaging method was applied to analysis of peripheral nerve regeneration by examining the sciatic nerve in vitro and in vivo. Raman spectra of intact nerve tissue had three particularly important peaks in the range 2800 − 3000 cm −1 . Spectra of injured sciatic nerves showed significant changes in the ratio of these peaks. Analysis of cellular spectra suggested that the spectrum for sciatic nerve tissue reflects the axon and myelin components of this tissue. Immunohistochemical analysis showed that the number of axons and the myelinated area were reduced at 7 days after injury and then increased by 28 days. The relative change in the axon to myelin ratio showed a similar initial increase, followed by a decrease at 28 days after injury. These changes correlated with the band intensity ratio and the changes in distribution of axon and myelin in Raman spectral analysis. Thus, our results suggest that label-free biochemical imaging with Raman spectroscopy can be used to detect turnover of axon and myelin in peripheral nerve regeneration. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Section: Discussionmentioning

confidence: 56%

Application of Raman spectroscopy for visualizing biochemical changes during peripheral nerve injuryin vitroandin vivo

et al. 2013

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“…Figure 4 shows the time-lapse MCDI of a single dividing Schizosaccharomyces pombe, fission yeast cell. 61 Raman mapping measurements were performed at 600 to 800 points (depending on the image size) at an interval of 0.5 μm and at nine different times (1, 2, 4, 6, 6.5, 10, 14, 18, and 22 h after inoculation of yeast cells into medium) in the cell cycle. The resultant 6885 Raman spectra were assembled to construct one A matrix; two spatial and one temporal dimensions were combined to a single dimension.…”

Section: Analysis Of Living Cellsmentioning

confidence: 99%

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

Ando

Hamaguchi

2013

J. Biomed. Opt

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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, timeand 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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“…Moreover, in combination with optical microscopy, Raman microspectroscopy provides high space-resolved information of human cells [19], fungi [20], or bacteria, including Streptomyces species [21]. Our research group has carried out time- and space-resolved Raman imaging of living yeast cells using confocal Raman microspectroscopy [22,23,24]. The Raman images of cells show that the distribution of lipids and proteins vary during cell division cycles.…”

Section: Introductionmentioning

confidence: 99%

In Situ Detection of Antibiotic Amphotericin B Produced in Streptomyces nodosus Using Raman Microspectroscopy

et al. 2014

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The study of spatial distribution of secondary metabolites within microbial cells facilitates the screening of candidate strains from marine environments for functional metabolites and allows for the subsequent assessment of the production of metabolites, such as antibiotics. This paper demonstrates the first application of Raman microspectroscopy for in situ detection of the antifungal antibiotic amphotericin B (AmB) produced by actinomycetes—Streptomyces nodosus. Raman spectra measured from hyphae of S. nodosus show the specific Raman bands, caused by resonance enhancement, corresponding to the polyene chain of AmB. In addition, Raman microspectroscopy enabled us to monitor the time-dependent change of AmB production corresponding to the growth of mycelia. The Raman images of S. nodosus reveal the heterogeneous distribution of AmB within the mycelia and individual hyphae. Moreover, the molecular association state of AmB in the mycelia was directly identified by observed Raman spectral shifts. These findings suggest that Raman microspectroscopy could be used for in situ monitoring of antibiotic production directly in marine microorganisms with a method that is non-destructive and does not require labeling.

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Disentangling Dynamic Changes of Multiple Cellular Components during the Yeast Cell Cycle by in Vivo Multivariate Raman Imaging

Cited by 81 publications

References 47 publications

Application of Raman spectroscopy for visualizing biochemical changes during peripheral nerve injuryin vitroandin vivo

Application of Raman spectroscopy for visualizing biochemical changes during peripheral nerve injuryin vitroandin vivo

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

In Situ Detection of Antibiotic Amphotericin B Produced in Streptomyces nodosus Using Raman Microspectroscopy

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