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分析方法，使我们能够测量 细胞对各种细胞外刺激的实时细胞内反应。我们方法的独特性依赖于 在几项创新上。我们将构建一个基于傅里叶变换的倒谱红外显微镜(ISIM)。 傅里叶变换红外光谱及其与新型等离子体生物传感器的结合 纳米结构(超表面)。该生物传感器将与多孔板集成在一起，以实现高分辨率 吞吐量。建议的化验将检测细胞的生化和形态变化， 强调细胞膜及其细胞骨架的重组。的多维性 光谱数据将使机器学习技术的应用成为可能，并提高其在 与传统的一维实时检测方法进行比较。据我们所知，这将是第一次实时 满足以下所有四个要求的细胞分析：(I)高通量，(Ii)多维性 收集的与时间相关的数据，(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。该化验将使用非常常见的 细胞的外部刺激，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.