SUMMARY Tools capable of imaging and perturbing mechanical signaling pathways with fine spatiotemporal resolution have been elusive despite their importance in diverse cellular processes. The challenge in developing a mechanogenetic toolkit (i.e. selective and quantitative activation of genetically encoded mechanoreceptors) stems from the fact that many mechanically-activated processes are localized in space and time, yet additionally require mechanical loading to become activated. To address this challenge, we synthesized magnetoplasmonic nanoparticles that can image, localize, and mechanically load targeted proteins with high spatiotemporal resolution. We demonstrate their utility by investigating the cell surface activation of two mechanoreceptors: Notch and E-cadherin. By measuring cellular responses to a spectrum of spatial, chemical, temporal, and mechanical inputs at the single molecule and single cell level, we reveal how spatial segregation and mechanical force cooperate to direct receptor activation dynamics. This generalizable technique can be used to control and understand diverse mechanosensitive processes in cell signaling.

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