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

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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.