Aberration-free FTIR spectroscopic imaging of live cells in microfluidic devices
Aberration-free FTIR spectroscopic imaging of live cells in microfluidic devices
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DOI：
10.1039/c3an00327b
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发表时间：
2013-01-01
期刊：
影响因子：
4.2
通讯作者：
Kazarian, Sergei G.
中科院分区：
文献类型：
--
作者：
Chan, K. L. Andrew;Kazarian, Sergei G.
FTIR光谱成像所提供的无标记、非破坏性的化学分析是研究活生物细胞的一个非常有吸引力和潜在的强大工具。活细胞的FTIR成像是一项具有挑战性的任务，因为细胞是在水性环境中培养的。虽然同步加速器设施已被证明是一个有价值的工具，单活细胞的FTIR显微光谱研究，我们已经证明，高品质的单活细胞的红外光谱，使用普通的Globar源也可以通过添加一对透镜，以一个共同的传输液体细胞。当放置在透射室窗口上时，透镜形成伪半球，其去除光的折射，从而改善所获得的数据的成像和光谱质量。本研究表明，可以获得单个活细胞的红外光谱，而不会产生色差引起的不同波数的焦移效应。单细胞的光谱已经证实，测量的光谱区域在整个范围内保持聚焦，而在没有透镜的情况下测量的单细胞的光谱由于焦点的偏移而显示出一些错误的特征。还已经证明，透镜的添加可以应用于微制造装置中的细胞的成像。我们已经表明，除非应用透镜，否则不可能获得油中DPBS液滴中孤立细胞的聚焦图像。使用本文所述的方法允许获得DPBS液滴中的单细胞的良好聚焦图像。
The label-free, non-destructive chemical analysis offered by FTIR spectroscopic imaging is a very attractive and potentially powerful tool for studies of live biological cells. FTIR imaging of live cells is a challenging task, due to the fact that cells are cultured in an aqueous environment. While the synchrotron facility has proven to be a valuable tool for FTIR microspectroscopic studies of single live cells, we have demonstrated that high quality infrared spectra of single live cells using an ordinary Globar source can also be obtained by adding a pair of lenses to a common transmission liquid cell. The lenses, when placed on the transmission cell window, form pseudo hemispheres which removes the refraction of light and hence improve the imaging and spectral quality of the obtained data. This study demonstrates that infrared spectra of single live cells can be obtained without the focus shifting effect at different wavenumbers, caused by the chromatic aberration. Spectra of the single cells have confirmed that the measured spectral region remains in focus across the whole range, while spectra of the single cells measured without the lenses have shown some erroneous features as a result of the shift of focus. It has also been demonstrated that the addition of lenses can be applied to the imaging of cells in microfabricated devices. We have shown that it was not possible to obtain a focused image of an isolated cell in a droplet of DPBS in oil unless the lenses are applied. The use of the approach described herein allows for well focused images of single cells in DPBS droplets to be obtained.