HiExM: high-throughput expansion microscopy enables scalable super-resolution imaging.

Expansion microscopy (ExM) involves the physical, isotropic expansion of specimens enabling nanoscale imaging using a standard confocal microscope. ExM is widely employed to image proteins, nucleic acids, and lipid membranes in single cells at nanoscale; however, current methods are low throughput which limits the number of samples that can be processed simultaneously. We developed High-throughput Expansion Microscopy (HiExM), a robust platform that enables expansion microscopy of cells cultured in a standard 96-well plate. Our method enables consistent ∼4.2x expansion across individual wells and entire plates in parallel. We also demonstrate that samples expanded using HiExM can be imaged and analyzed using standard high-throughput confocal imaging platforms. Our method broadens the application of ExM as a tool for scalable super-resolution imaging of specimens for pharmaceutical and research applications including drug and cell phenotype screening.