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Increased optical pathlength through aqueous media for the infrared microanalysis of live cells
Analytical and Bioanalytical Chemistry ( IF 4.3 ) Pub Date : 2018-07-02 , DOI: 10.1007/s00216-018-1188-2
James Doherty , Zhe Zhang , Katia Wehbe , Gianfelice Cinque , Peter Gardner , Joanna Denbigh

The study of live cells using Fourier transform infrared spectroscopy (FTIR) and FTIR microspectroscopy (FT-IRMS) intrinsically yields more information about cell metabolism than comparable experiments using dried or chemically fixed samples. There are, however, a number of barriers to obtaining high-quality vibrational spectra of live cells, including correction for the significant contributions of water bands to the spectra, and the physical stresses placed upon cells by compression in short pathlength sample holders. In this study, we present a water correction method that is able to result in good-quality cell spectra from water layers of 10 and 12 μm and demonstrate that sufficient biological detail is retained to separate spectra of live cells based upon their exposure to different novel anti-cancer agents. The IR brilliance of a synchrotron radiation (SR) source overcomes the problem of the strong water absorption and provides cell spectra with good signal-to-noise ratio for further analysis. Supervised multivariate analysis (MVA) and investigation of average spectra have shown significant separation between control cells and cells treated with the DNA cross-linker PL63 on the basis of phosphate and DNA-related signatures. Meanwhile, the same control cells can be significantly distinguished from cells treated with the protein kinase inhibitor YA1 based on changes in the amide II region. Each of these separations can be linked directly to the known biochemical mode of action of each agent.

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中文翻译:

通过水性介质的光程增加,用于活细胞的红外微分析

与使用干燥或化学固定样品的可比实验相比,使用傅里叶变换红外光谱(FTIR)和FTIR显微光谱(FT-IRMS)进行的活细胞研究本质上可提供有关细胞代谢的更多信息。但是,要获得活细胞的高质量振动光谱,存在许多障碍,包括校正水带对光谱的重要贡献以及在短光程样品架中通过压缩作用在细胞上的物理应力。在这项研究中,我们提出了一种水校正方法,该方法能够从10和12μm的水层中产生高质量的细胞光谱,并证明保留了足够的生物学细节,可以根据活细胞暴露于不同的新型物质而分离出活细胞的光谱。抗癌药。同步辐射(SR)源的IR亮度克服了强吸水率的问题,并为细胞光谱提供了良好的信噪比,以供进一步分析。监督多变量分析(MVA)和平均光谱研究表明,在磷酸盐和DNA相关信号的基础上,对照细胞与用DNA交联剂PL63处理的细胞之间存在明显的分离。同时,基于酰胺II区域的变化,可以将相同的对照细胞与用蛋白激酶抑制剂YA1处理的细胞区分开。这些分离中的每一个都可以直接与每种试剂的已知生化作用模式联系起来。监督多变量分析(MVA)和平均光谱研究表明,在磷酸盐和DNA相关信号的基础上,对照细胞与用DNA交联剂PL63处理的细胞之间存在明显的分离。同时,基于酰胺II区域的变化,可以将相同的对照细胞与用蛋白激酶抑制剂YA1处理的细胞区分开。这些分离中的每一个都可以直接与每种试剂的已知生化作用模式联系起来。监督多变量分析(MVA)和平均光谱研究表明,在磷酸盐和DNA相关信号的基础上,对照细胞与用DNA交联剂PL63处理的细胞之间存在明显的分离。同时,基于酰胺II区域的变化,可以将相同的对照细胞与用蛋白激酶抑制剂YA1处理的细胞区分开。这些分离中的每一个都可以直接与每种试剂的已知生化作用模式联系起来。基于酰胺II区的变化,可以将相同的对照细胞与用蛋白激酶抑制剂YA1处理的细胞区分开来。这些分离中的每一个都可以直接与每种试剂的已知生化作用模式联系起来。基于酰胺II区的变化,可以将相同的对照细胞与用蛋白激酶抑制剂YA1处理的细胞区分开来。这些分离中的每一个都可以直接与每种试剂的已知生化作用模式联系起来。

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更新日期:2018-07-02
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