Non-contact mechanical and chemical analysis of single living cells by microspectroscopic techniques
Non-contact mechanical and chemical analysis of single living cells by microspectroscopic techniques
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DOI：
10.1038/lsa.2017.139
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发表时间：
2018-02-09
影响因子：
19.4
通讯作者：
Caponi, Silvia
中科院分区：
文献类型：
--
作者：
Mattana, Sara;Mattarelli, Maurizio;Caponi, Silvia
创新的无标签微光谱学可以同时收集布里布鲁和拉曼信号，用于以亚微米分辨率表征活细胞的粘弹性特性和化学成分。前所未有的数据统计精度，结合高频分辨率和最近建立的实验装置的高对比度，允许通过非接触测量研究浸泡在缓冲溶液中的单个活细胞。布里渊信号在缓冲液和细胞成分中反卷积，从而揭示细胞内部的机械非均质性。特别是，通过与拉曼光谱标记比较，观察到从质膜到核的弹性模量增加了20%。布里渊线形状分析更适用于生理和病理条件下细胞的比较。癌基因表达后，细胞整体弹性模量（15%）和表观黏度（50%）降低。在一个原理验证实验中，这种光谱技术表征亚细胞区室和区分细胞状态的能力已成功测试。该结果有力地支持了该技术在生物医学领域基础问题上的未来应用。
Innovative label-free microspectroscopy, which can simultaneously collect Brillouin and Raman signals, is used to characterize the viscoelastic properties and chemical composition of living cells with sub-micrometric resolution. The unprecedented statistical accuracy of the data combined with the high-frequency resolution and the high contrast of the recently built experimental setup permits the study of single living cells immersed in their buffer solution by contactless measurements. The Brillouin signal is deconvoluted in the buffer and the cell components, thereby revealing the mechanical heterogeneity inside the cell. In particular, a 20% increase is observed in the elastic modulus passing from the plasmatic membrane to the nucleus as distinguished by comparison with the Raman spectroscopic marker. Brillouin line shape analysis is even more relevant for the comparison of cells under physiological and pathological conditions. Following oncogene expression, cells show an overall reduction in the elastic modulus (15%) and apparent viscosity (50%). In a proof-of-principle experiment, the ability of this spectroscopic technique to characterize subcellular compartments and distinguish cell status was successfully tested. The results strongly support the future application of this technique for fundamental issues in the biomedical field.