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Nano Lett. 2019 Apr 10;19(4):2641-2646. doi: 10.1021/acs.nanolett.9b00508. Epub 2019 Mar 13.

124-Color Super-resolution Imaging by Engineering DNA-PAINT Blinking Kinetics.

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Department of Physics and Center for Nanoscience , Ludwig Maximilian University , 80539 Munich , Germany.
Max Planck Institute of Biochemistry , 82152 Martinsried , Germany.
Wyss Institute for Biologically Inspired Engineering , Harvard University , Boston , Massachusetts 02138 , United States.
Department of Systems Biology , Harvard Medical School , Boston , Massachusetts 02115 , United States.


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.


DNA nanotechnology; DNA-PAINT; Super-resolution microscopy; barcoding; multiplexing

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