An Integrated Platform for High-Throughput Nanoscopy

• Published in: bioRxiv
• Main Authors: Barentine, Andrew E S, Lin, Yu, Courvan, Edward M, Kidd, Phylicia, Liu, Miao, Balduf, Leonhard, Phan, Timy, Rivera-Molina, Felix, Grace, Michael R, Marin, Zach, Lessard, Mark, Rios-Chen, Juliana, Wang, Siyuan, Neugebauer, Karla M, Bewersdorf, Joerg, Baddeley, David
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 20.04.2022; Cold Spring Harbor Laboratory
• Edition: 1.3
• Subjects: Biophysics; Data acquisition; Data processing; Mammalian cells
• ISSN: 2692-8205; 2692-8205
• 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. Competing Interest Statement ificant financial interest in Bruker Corp. and Hamamatsu Photonics Footnotes * Added new biological application of method.