Analysis of Fixed and Live Single Cells Using Optical Photothermal Infrared with Concomitant Raman Spectroscopy.

Analysis of Fixed and Live Single Cells Using Optical Photothermal Infrared with Concomitant Raman Spectroscopy.
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DOI:
10.1021/acs.analchem.0c04846
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发表时间:
2021-03-02
影响因子:
7.4
通讯作者:
Gardner P
Gardner P
中科院分区:
化学1区
文献类型:
--
作者:
Spadea A;Denbigh J;Lawrence MJ;Kansiz M;Gardner P

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本文报道了首次使用一种全新的完全基于光学的光热方法(O-PTIR),利用量子级联激光器(QCL)和光学参量振荡器(OPO)激光器作为激发源,获得固定细胞和活细胞的红外光谱,从而使所有与生物相关的振动能够在亚微米空间分辨率下进行分析。此外,红外数据采集与来自完全相同激发位置的伴随拉曼光谱相结合,这意味着可以获得细胞的完整振动剖面。胰腺癌细胞系MIA PaCa-2和乳腺癌细胞系MDA-MB-231被用作模型细胞,以展示新仪器的功能。这些组合的模式可以用于分析固定的亚细胞结构,更重要的是,在水条件下的活细胞。我们发现蛋白质二级结构和富脂体可以在亚微米尺度上识别。
This paper reports the first use of a novel completely optically based photothermal method (O-PTIR) for obtaining infrared spectra of both fixed and living cells using a quantum cascade laser (QCL) and optical parametric oscillator (OPO) laser as excitation sources, thus enabling all biologically relevant vibrations to be analyzed at submicron spatial resolution. In addition, infrared data acquisition is combined with concomitant Raman spectra from exactly the same excitation location, meaning the full vibrational profile of the cell can be obtained. The pancreatic cancer cell line MIA PaCa-2 and the breast cancer cell line MDA-MB-231 are used as model cells to demonstrate the capabilities of the new instrumentation. These combined modalities can be used to analyze subcellular structures in both fixed and, more importantly, live cells under aqueous conditions. We show that the protein secondary structure and lipid-rich bodies can be identified on the submicron scale.
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