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We present a parallel stimulated emission depletion (STED) nanoscope with no mechanical moving parts and sub-millisecond pixel dwell times, relying on electro-optical (EO) phase modulators. The nanoscope offers 1225-fold parallelization over single-doughnut-scanning STED and achieves a spatial resolution of 35 nm. We imaged immunostained nuclear pore complexes of zebrafish within their natural biological environment, demonstrating spatial and temporal resolutions of 56 nm and 0.2 s, respectively. Furthermore, we show parallel EO-STED sub-second imaging of microtubules inside living cells. Finally, we reveal the nanodomain organization of a eukaryotic initiation factor within the processing bodies of fixed cells. The potential of parallel EO-STED to offer microsecond pixel dwell times over large fields of view promises millisecond STED imaging.

Sub-second, super-resolved imaging of biological systems using parallel EO-STED