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Left-handed DNA for efficient highly multiplexed imaging at single-protein resolution
Eduard M Unterauer1,2, Eva-Maria Schentarra1,2, Isabelle Pachmayr2,3
1Faculty of Physics and Center for Nanoscience, Ludwig Maximilian University, Munich, Germany.
Nature Communications
|October 2, 2025
Summary
Researchers developed a faster 12-plex imaging method using DNA-PAINT for spatial proteomics. This technique achieves high-resolution mapping of neuronal protein interactions in 3D, overcoming limitations of previous methods.
Area of Science:
- Molecular Biology
- Microscopy
- Proteomics
Background:
- Multiplexed super-resolution microscopy allows spatial proteomics at the single-protein level.
- Existing methods often require secondary labels, which complicates workflows and reduces throughput.
- There is a need for streamlined, high-throughput spatial proteomics techniques.
Purpose of the Study:
- To introduce a streamlined approach for rapid and efficient multiplexed super-resolution imaging.
- To enable 12-plex imaging with high spatial resolution and throughput.
- To map dense neuronal interactomes in 3D.
Main Methods:
- Development of a streamlined method combining speed-optimized DNA-PAINT sequences.
- Utilization of mirror-image DNA analogs (left-handed DNA) for enhanced imaging.
- Validation on synthetic and cellular benchmarks.
Main Results:
- Achieved rapid and efficient 12-plex imaging.
- Demonstrated mapping of dense neuronal interactomes in three dimensions.
- Obtained 15 nm spatial resolution over a 200 × 200 µm² field of view.
Conclusions:
- The developed method significantly improves the efficiency and throughput of spatial proteomics.
- This technique provides high-resolution 3D mapping of protein interactions in complex biological systems.
- The streamlined approach simplifies implementation compared to existing multiplexed microscopy methods.
