High-resolution imaging mass cytometry to map subcellular structures

Imaging mass cytometry (IMC) is a powerful multiplexed imaging technology used to investigate cell phenotypes and spatial organization of tissue in health and disease. The spatial resolution of IMC is presently at 1 µm, enabling the resolution of single cells and large subcellular compartments but not submicrometer intracellular structures. Here we report a method to improve the resolution of IMC so that it approaches that of light microscopy. High-resolution IMC (HR-IMC) uses an oversampling approach coupled with point-spread function-based deconvolution to achieve a resolution below 350 nm. We demonstrate the performance of HR-IMC in resolving subcellular structures, such as nuclear foci and mitochondrial networks previously undetectable with IMC, and applied it to visualize chemotherapy-induced perturbation of patient-derived ovarian cancer cells. HR-IMC extends highly multiplex IMC analyses into the subcellular regime, enabling analysis of cell biological features and characteristics of disease.