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Schueder, Florian; Lara-Gutierrez, Juanita; Haas, Daniel; Beckwith, Kai Sandvold; Yin, Peng; Ellenberg, Jan; Jungmann, Ralf (2020): Super-Resolution Spatial Proximity Detection with Proximity-PAINT. In: Angewandte Chemie-International Edition, Vol. 60, No. 2: pp. 716-720
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Visualizing the functional interactions of biomolecules such as proteins and nucleic acids is key to understanding cellular life on the molecular scale. Spatial proximity is often used as a proxy for the direct interaction of biomolecules. However, current techniques to visualize spatial proximity are either limited by spatial resolution, dynamic range, or lack of single-molecule sensitivity. Here, we introduce Proximity-PAINT (pPAINT), a variation of the super-resolution microscopy technique DNA-PAINT. pPAINT uses a split-docking-site configuration to detect spatial proximity with high sensitivity, low false-positive rates, and tunable detection distances. We benchmark and optimize pPAINT using designer DNA nanostructures and demonstrate its cellular applicability by visualizing the spatial proximity of alpha- and beta-tubulin in microtubules using super-resolution detection.