用于研究人类粒细胞的拉曼显微光谱方法。
Raman microspectroscopic approach to the study of human granulocytes.
作者信息
Puppels G J, Garritsen H S, Segers-Nolten G M, de Mul F F, Greve J
机构信息
Biophysical Technology Group, Faculty of Applied Physics, University of Twente, Enschede, The Netherlands.
出版信息
Biophys J. 1991 Nov;60(5):1046-56. doi: 10.1016/S0006-3495(91)82142-7.
A sensitive confocal Raman microspectrometer was employed to record spectra of nuclei and cytoplasmic regions of single living human granulocytes. Conditions were used that ensured cell viability and reproducibility of the spectra. Identical spectra were obtained from the nuclei of neutrophilic, eosinophilic, and basophilic granulocytes, which yield information about DNA and protein secondary structure and DNA-protein ratio. The cytoplasmic Raman spectra of the three cell types are very different. This was found to be mainly due to the abundant presence of peroxidases in the cytoplasmic granules of neutrophilic granulocytes (myeloperoxidase) and eosinophilic granulocytes (eosinophil peroxidase). Strong signal contributions of the active site heme group(s) of these enzymes were found. This paper illustrates the potentials and limitations for Raman spectroscopic analysis of cellular constituents and processes.
使用一台灵敏的共聚焦拉曼光谱仪记录单个活的人类粒细胞的细胞核和细胞质区域的光谱。采用了确保细胞活力和光谱可重复性的条件。从中性粒细胞、嗜酸性粒细胞和嗜碱性粒细胞的细胞核获得了相同的光谱，这些光谱提供了有关DNA和蛋白质二级结构以及DNA-蛋白质比率的信息。三种细胞类型的细胞质拉曼光谱非常不同。发现这主要是由于嗜中性粒细胞（髓过氧化物酶）和嗜酸性粒细胞（嗜酸性粒细胞过氧化物酶）的细胞质颗粒中大量存在过氧化物酶。发现了这些酶的活性位点血红素基团的强烈信号贡献。本文阐述了拉曼光谱分析细胞成分和过程的潜力和局限性。
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