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3D Multicolor Nanoscopy at 10,000 Cells a Day

By Andrew E. S. Barentine, Yu Lin, Miao Liu, Phylicia Kidd, Leonhard Balduf, Michael R. Grace, Siyuan Wang, Joerg Bewersdorf, David Baddeley

Posted 12 Apr 2019
bioRxiv DOI: 10.1101/606954

Diffraction-unlimited single-molecule switching (SMS) nanoscopy techniques like STORM /(F)PALM enable three-dimensional (3D) fluorescence imaging at 20-80 nm resolution and are invaluable to investigate sub-cellular organization. They suffer, however, from low throughput, limiting the output of a day's worth of imaging to typically a few tens of mammalian cells. Here we develop an SMS imaging platform that combines high-speed 3D single-molecule data acquisition with an automated, fully integrated, high-volume data processing pipeline. We demonstrate 2-color 3D super-resolution imaging of over 10,000 mammalian cell nuclei in about 26 hours, connecting the traditionally low-throughput super-resolution community to the world of omics approaches.

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