Swiss Light Source, Paul Scherrer Institut , CH-5232 Villigen, Switzerland.
Anal Chem. 2014 Jul 15;86(14):6887-95. doi: 10.1021/ac501561x. Epub 2014 Jun 26.
In recent years, major efforts have been devoted to the application of microscopy with mid-infrared light to the study of living cells and tissue. Despite this interest, infrared (IR) microscopy has not realized its full potential in the molecular characterization of living systems. This is partly due to the fact that current approaches for data mining and analysis of IR absorption spectra have not evolved comparably to measurement technology and are not up to the interpretation of the complex spectra of living systems such as cells and tissue. In this work we show that the use of two-dimensional correlation spectroscopy coupled to IR absorption spectro-microscopy allows us to extract the spectral components of individual metabolites from time-resolved IR spectra of living cells. We call this method correlated cellular spectro-microscopy, and we implement it in the study of the glycolytic metabolism of cancer cells. We show that the method can detect intermediates of the glycolytic pathway, quantify their rate of formation, and correlate this with variations in pH, all in a single measurement. We propose the method as a useful tool for the quantitative description of metabolic processes in living cells and for the validation of drug candidates aimed at suppressing glycolysis in cancer cells.
近年来，人们致力于将中红外光显微镜应用于活细胞和组织的研究。尽管人们对此很感兴趣，但在活细胞体系的分子特征分析中，红外（IR）显微镜并未充分发挥其潜力。这部分是因为目前用于挖掘和分析 IR 吸收光谱的数据的方法并没有与测量技术同步发展，无法对细胞和组织等复杂的活细胞体系的光谱进行解读。在这项工作中，我们表明，将二维相关光谱学与 IR 吸收光谱显微镜结合使用，可以从活细胞的时间分辨 IR 光谱中提取单个代谢物的光谱分量。我们将这种方法称为相关细胞光谱显微镜，并将其应用于癌细胞糖酵解代谢的研究。我们表明，该方法可以检测糖酵解途径的中间产物，定量它们的形成速率，并将其与 pH 值的变化相关联，所有这些都可以在一次测量中完成。我们提出该方法作为一种定量描述活细胞代谢过程和验证旨在抑制癌细胞糖酵解的候选药物的有用工具。
