Inverted Spectroscopic Infrared Microscope (ISIM) for High Throughput Multi-Dimensional Cell Assays

摘要 这项研究的目标是开发一种新的光谱细胞测定法，使我们能够测量细胞内的 细胞对各种细胞外刺激的实时细胞内反应。我们方法的独特性在于 几项创新。我们将建立一个倒置光谱红外显微镜（ISIM）的基础上傅立叶- 变换红外（FTIR）光谱，并将其与基于等离子体激元的新型生物传感器相结合， 纳米结构（超颖表面）。生物传感器将与多孔板集成，以实现高- 吞吐量所提出的测定将检测细胞的生物化学和形态学变化， 强调细胞膜及其细胞骨架的重组。的多层面性质 光谱数据将使机器学习技术的应用成为可能，并提高其灵敏度， 与传统的一维实时分析比较。据我们所知，这将是第一个实时的 满足以下所有四个要求的细胞测定：（i）高通量，（ii）多维度的 收集的时间相关数据，（iii）具体关注细胞的生化变化，以及（iv）关注 发生在细胞膜附近的变化。将使用非常常见的 细胞的外部刺激，例如作用于G蛋白偶联受体的小分子化合物。

Abstract The goal of this research is to develop a novel spectroscopic cellular assay that will enable us to measure the real-time intra-cellular response of a cell to various extra-cellular stimuli. The uniqueness of our approach relies on several innovations. We will construct an Inverted Spectral Infrared Microscope (ISIM) based on Fourier- transform infrared (FTIR) spectroscopy and combining it with a novel biosensor based on plasmonic nanostructures (metasurfaces). The biosensor will be integrated with multi-well plates to enable high- throughput. The proposed assay will detect biochemical and morphological changes of the cell, with the emphasis on the reorganization of the cellular membrane and its cytoskeleton. The multi-dimensional nature of the spectroscopic data will enable the application of machine learning techniques and improve its sensitivity in comparison with traditional one-dimensional real-time assays. To our knowledge, this will be the first real-time cellular assay that satisfies all of the four requirements below: (i) high throughput, (ii) multi-dimensionality of the collected time-dependent data, (iii) specific focus on biochemical changes of the cell, and (iv) focus on the changes occurring in close proximity of the cellular membrane. The assay will be validated using very common external stimuli of the cell, such as small-molecule compounds acting on G-protein coupled receptors.