Huang Steven H, Shen Po-Ting, Mahalanabish Aditya, Sartorello Giovanni, Shvets Gennady
bioRxiv. 2024 Sep 18:2024.09.17.613596. doi: 10.1101/2024.09.17.613596.
Mid-Infrared (MIR) chemical imaging provides rich chemical information of biological samples in a label-free and non-destructive manner. Yet, its adoption to live-cell analysis is limited by the strong attenuation of MIR light in water, often necessitating cell culture geometries that are incompatible with the prolonged viability of cells and with standard high-throughput workflow. Here, we introduce a new approach to MIR microscopy, where cells are imaged through their localized near-field interaction with a plasmonic metasurface. Chemical contrast of distinct molecular groups provided sub-cellular resolution images of the proteins, lipids, and nucleic acids in the cells that were collected using an inverted MIR microscope. Time-lapse imaging of living cells demonstrated that their behaviors, including motility, viability, and substrate adhesion, can be monitored over extended periods of time using low-power MIR light. The presented approach provides a method for the non-perturbative MIR imaging of living cells, which is well-suited for integration with modern high-throughput screening technologies for the label-free, high-content chemical imaging of living cells.
中红外（MIR）化学成像能够以无标记且非破坏性的方式提供生物样品丰富的化学信息。然而，其在活细胞分析中的应用受到MIR光在水中强烈衰减的限制，这通常需要采用与细胞长期存活以及标准高通量工作流程不兼容的细胞培养几何结构。在此，我们介绍一种MIR显微镜的新方法，即通过细胞与等离子体超表面的局部近场相互作用对细胞进行成像。利用倒置MIR显微镜采集的不同分子基团的化学对比度提供了细胞中蛋白质、脂质和核酸的亚细胞分辨率图像。活细胞的延时成像表明，使用低功率MIR光可以长时间监测它们的行为，包括运动性、活力和底物粘附。所提出的方法提供了一种对活细胞进行非侵入性MIR成像的方法，非常适合与现代高通量筛选技术集成，用于活细胞的无标记、高内涵化学成像。
