Molecular-level investigation of the structure, transformation, and bioactivity of single living fission yeast cells by time- and space-resolved Raman spectroscopy
Molecular-level investigation of the structure, transformation, and bioactivity of single living fission yeast cells by time- and space-resolved Raman spectroscopy
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DOI：
10.1021/bi050179w
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发表时间：
2005-08-02
期刊：
影响因子：
2.9
通讯作者：
Hamaguchi, H
中科院分区：
文献类型：
--
作者：
Huang, YS;Karashima, T;Hamaguchi, H
利用时间和空间分辨的拉曼光谱技术，在分子水平上研究了粟酒裂殖酵母单个活细胞的结构、转化和生物活性。共焦拉曼显微光谱仪的时间分辨率为100 s，空间分辨率为250 nm。用空间分辨拉曼光谱研究了活S.记录了不同细胞周期阶段的粟酒酵母细胞，试图阐明细胞器的分子组成，包括细胞核、细胞质、线粒体和隔膜。分裂的酵母细胞的中央部分的时间和空间分辨的测量显示连续的光谱演化从细胞核的细胞质和线粒体，最后到隔膜，根据在细胞周期中的转换。只有当细胞处于良好的营养条件下时，才在1602 cm（-1）处观察到强的拉曼谱带。通过测量酵母细胞线粒体的拉曼光谱，研究了呼吸抑制剂KCN对酵母细胞的影响。观察到1602 cm（-1）带突然消失，随后磷脂带的形状和强度发生变化，表明细胞活性与该带的强度之间存在密切关系。因此，我们把这个波段称为“生命的拉曼光谱特征”。还进行了活酵母细胞的拉曼映射。不仅分子种类的分布，而且那些活跃的线粒体在细胞中被成功地可视化在体内。
The structure, transformation, and bioactivity of single living Schizosaccharomyces pombe cells at the molecular level have been studied in vivo by time- and space-resolved Raman spectroscopy. A time resolution of 100 s and a space resolution of 250 nm have been achieved with the use of a confocal Raman microspectrometer. The space-resolved Raman spectra of living S. pombe cells at different cell cycle stages were recorded in an effort to elucidate the molecular compositions of organelles, including nuclei, cytoplasm, mitochondria, and septa. The time- and space-resolved measurement of the central part of a dividing yeast cell showed continuous spectral evolution from that of the nucleus to those of the cytoplasm and mitochondria and finally to that of the septum, in accordance with the transformation during the cell cycle. A strong Raman band was observed at 1602 cm(-1) only when cells were under good nutrient conditions. The effect of a respiration inhibitor, KCN, on a living yeast cell was studied by measuring the Raman spectra of its mitochondria. A sudden disappearance of the 1602 cm(-1) band followed by the change in the shape and intensity of the phospholipid bands was observed, indicating a strong relationship between the cell activity and the intensity of this band. We therefore call this band "the Raman spectroscopic signature of life". The Raman mapping of a living yeast cell was also carried out. Not only the distributions of molecular species but also those of active mitochondria in the cell were successfully visualized in vivo.