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 () and Ralf Jungmann ()
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Eduard M. Unterauer: Ludwig Maximilian University
Eva-Maria Schentarra: Ludwig Maximilian University
Isabelle Pachmayr: Max Planck Institute of Biochemistry
Taisha Tashrin: Max Planck Institute of Biochemistry
Jisoo Kwon: Ludwig Maximilian University
Sebastian Strauss: Max Planck Institute of Biochemistry
Kristina Jevdokimenko: University Medical Center Göttingen
Rafal Kowalewski: Ludwig Maximilian University
Felipe Opazo: University Medical Center Göttingen
Eugenio F. Fornasiero: University Medical Center Göttingen
Luciano A. Masullo: Max Planck Institute of Biochemistry
Ralf Jungmann: Ludwig Maximilian University

Nature Communications, 2025, vol. 16, issue 1, 1-8

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 µm2 field of view.

Date: 2025
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DOI: 10.1038/s41467-025-64228-x

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