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Left-handed DNA for efficient highly multiplexed imaging at single-protein resolution

Eduard M. Unterauer, Eva-Maria Schentarra, Isabelle Pachmayr, Taisha Tashrin, Jisoo Kwon, Sebastian Strauss, Kristina Jevdokimenko, Rafal Kowalewski, Felipe Opazo, Eugenio F. Fornasiero, Luciano A. Masullo, Ralf Jungmann

2025Nature Communications5 citationsDOIOpen Access PDF

Abstract

Abstract Multiplexed super-resolution microscopy enables spatial proteomics at single-protein resolution, but current methods often depend on secondary labels, complicating implementation and limiting throughput. We introduce a streamlined approach that combines speed-optimized DNA-PAINT sequences with their mirror-image analogs (left-handed DNA), enabling rapid and efficient 12-plex imaging. Validated on synthetic and cellular benchmarks, our method maps dense neuronal interactomes in 3D with 15 nm spatial resolution across a 200 × 200 µm 2 field of view.

Topics & Concepts

LimitingMultiplexingComputer scienceResolution (logic)Field (mathematics)MicroscopyDNAImage resolutionProteomicsComputational biologyNanotechnologyHigh resolutionSuperresolutionEscherichia coli ProteinsDNA sequencingAtomic force microscopyMaterials scienceCurrent (fluid)Biological systemAdvanced Fluorescence Microscopy TechniquesCell Image Analysis TechniquesAdvanced Electron Microscopy Techniques and Applications
Left-handed DNA for efficient highly multiplexed imaging at single-protein resolution | Litcius