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Wade, Orsolya K.; Woehrstein, Johannes B.; Nickels, Philipp C.; Strauss, Sebastian; Stehr, Florian; Stein, Johannes; Schueder, Florian; Strauss, Maximilian T.; Ganji, Mahipal; Schnitzbauer, Jörg; Grabmayr, Heinrich; Yin, Peng; Schwille, Petra; Jungmann, Ralf (2019): 124-Color Super-resolution Imaging by Engineering DNA-PAINT Blinking Kinetics. In: Nano Letters, Vol. 19, No. 4: pp. 2641-2646
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Abstract

Optical super-resolution techniques reach unprecedented spatial resolution down to a few nanometers. However, efficient multiplexing strategies for the simultaneous detection of hundreds of molecular species are still elusive. Here, we introduce an entirely new approach to multiplexed super-resolution microscopy by designing the blinking behavior of targets with engineered binding frequency and duration in DNA-PAINT. We assay this kinetic barcoding approach in silico and in vitro using DNA origami structures, show the applicability for multiplexed RNA and protein detection in cells, and finally experimentally demonstrate 124-plex super-resolution imaging within minutes.