We hope G5M pushes molecular mapping forward. Excited to see the discoveries it enables! Thanks to @rafalkowalew.bsky.social, @susannereinhardt.bsky.social, @ipachmayr.bsky.social, Shuhan Xu, @lumasullo.bsky.social for developing G5M and the @jungmannlab.bsky.social for testing and improving it. 7/7
G5M is implemented in Picasso since version 0.9.5: github.com/jungmannlab/..., although we always recommend using the most recent version! A short documentation: picassosr.readthedocs.io/en/latest/re.... We advise reading the whole paper for best results! 6/7
Bonus: we found a new formula for axial loc. precision for astigmatic imaging for any z position. It only depends on the 3D calibration and the properties of localizations. Interestingly, the results are not necessarily symmetric around z = 0. 5/7
We validated G5M on DNA origami, Nuclear Pore Complexes (NPCs) and Obinutuzumab-treated CD20 RESI data. In NPCs, G5M found twice as many Nup96 dimers as GA, demonstrating increased accuracy in molecular mapping. 4/7
G5M uses modified Gaussian mixture modeling to accurately capture the shape of 2D and 3D astigmatic localization clouds. In realistic simulations, it resolves molecules spaced 2.9 localization precisions apart with a 27× higher recovery rate than other tools. 3/7
Knowing where biomolecules are is key to uncovering molecular interactions and patterns. But standard clustering methods (e.g., DBSCAN, GA) often fail when proteins are spaced <5× the localization precision, leading to false negatives and misassignments. 2/7
In DNA-PAINT, nearby proteins create overlapping localizations. You may see two by eye, but analysis often misses them, wasting precious precision. G5M solves this. Now in @natcomms.nature.com. doi.org/10.1038/s414.... 1/7
Big congrats to Clemens Steinek, @ipachmayr.bsky.social, and Sebastian Strauss who led the project as well as other co-authors that contributed to this work!! 6/6
Importantly, this workflow is broadly applicable and compatible with virtually any high off-rate binder. Many “bad” binders currently sitting in lab freezers may now become powerful imaging tools. 5/6
Using DyBE, we resolved the organization of receptor tyrosine kinases at single-protein resolution and detected ligand-dependent homodimerization of HER2, as well as EGF-induced formation of EGFR homodimers and EGFR–HER2 heterodimers. 4/6
DyBE increases protein labeling up to 15-fold for high off-rate nanobodies, enabling visualization of most molecules of a given target protein within the cell. 3/6
In classical DNA-PAINT, small binders such as nanobodies can localize proteins with nanometer precision, but rapid unbinding often limits efficient protein labeling. DyBE adapts DNA-PAINT to harness small, high off-rate binders for nanometer-precise sampling. 2/6
Here we introduce Dynamic Binder Exchange (DyBE), a new strategy that uses the high off-rate kinetics of small binders to map proteins with nanometer-scale precision! 1/6
Up to 15-fold improvement in protein labeling for DNA-PAINT 🧬🔬. We are excited to present our latest work published in @angewandtechemie.bsky.social!
onlinelibrary.wiley.com/doi/10.1002/...
How do you scale super-resolution microscopy to dozens of proteins without linearly scaling imaging time?
We introduce Combi-PAINT: a combinatorial DNA-PAINT strategy that breaks the 1-target-per-round bottleneck of Exchange-PAINT: www.biorxiv.org/content/10.6...
1/5
Ralf Jungmann, of the @mpibiochem.bsky.social, is the first speaker to step to the BiOS Hot Topics stage at #PhotonicsWest!💡
He is delivering his presentation titled: “From DNA nanotechnology to biomedical insight: towards single-molecule spatial omics”
@jungmannlab.bsky.social
The image on the cover shows two sugars from the same cell-surface glycan separated by 9 Å, visualized with RESI (resolution enhancement by sequential imaging) enabled by metabolic labelling with DNA barcodes. IMAGE: Luciano A. Masullo, Max Planck Institute of Biochemistry, Germany. COVER DESIGN: Vanitha Selvarajan Original paper: Masullo, L.A., et al. Ångström-resolution imaging of cell-surface glycans. Nat. Nanotechnol. 20, 1457–1463 (2025). https://doi.org/10.1038/s41565-025-01966-5 Abstract: Glycobiology is rooted in the study of monosaccharides, ångström-sized molecules that are the building blocks of glycosylation. Glycosylated biomolecules form the glycocalyx, a dense coat encasing every human cell with central relevance—among others—in immunology, oncology and virology. To understand glycosylation function, visualizing its molecular structure is fundamental. However, the ability to visualize the molecular architecture of the glycocalyx has remained challenging. Techniques such as mass spectrometry, electron microscopy and fluorescence microscopy lack the necessary cellular context, specificity and resolution. Here we combine resolution enhancement by sequential imaging with metabolic labelling, enabling the visualization of individual sugars within glycans on the cell surface, thus obtaining images of the glycocalyx with a spatial resolution down to 9 Å in an optical microscope.
Now online: October 2025 Issue.
- Focus Issue on #biosensing,
- DNA moiré superlattices,
- Sugars at Ångström-resolution,
- Solid-state #nanopores,
- Non-aqueous Li #batteries, -
- Neuromorphic vision,
- Peptide #hydrogels,
- Deep learning for #LNPs and more...
www.nature.com/nnano/volume...
Thanks to all who made this possible! @eduardunterauer.bsky.social @evaschentarra.bsky.social @ipachmayr.bsky.social @taishatashrin.bsky.social Jisoo Kwon Sebastian Strauss, @jekristina.bsky.social @rafalkowalew.bsky.social @opazo.bsky.social @forna.bsky.social @lumasullo.bsky.social (6/6)
Within this neuronal atlas we can reveal the three synapse classes, excitatory, inhibitory and the recently discovered mixed synapse. Organelle imaging of Peroxisomes (Pmp70) and the Golgi Apparatus (Golga5) reveals rare contact sides and even fused particles. (5/6)
To show the power of the technique, we acquired a 13-plex 200 x 200 µm2 neuronal atlas in 3D. With this atlas we map the interaction architecture of three neurons, resolving organelles, cytoskeleton, vesicles and synapses at single-protein resolution. (4/6)
We demonstrate speed-optimized left-handed DNA-PAINT by characterizing the sequence binding kinetics and resolving three main microscopy benchmarking targets, mitochondria, microtubules and nuclear pore complexes with <5 nm localization precision. (3/6)
The mirrored design of left-handed oligonucleotides allows the extension of the common 6 speed-sequences R1-R6 with their analogs L1-L6, enabling 12 target multiplexing with a standard secondary label-free DNA-PAINT workflow. (2/6)
Highly efficient 12-color multiplexing with speed-optimized DNA-PAINT. We are excited to share our latest paper in @natcomms.nature.com, using left-handed DNA to extend speed-optimized DNA-PAINT to 12 targets in a simple and straightforward way! 🧬👈🚀https://www.nature.com/articles/s41467-025-64228-x
Next on stage is Eduard Unterauer @eduardunterauer.bsky.social from @jungmannlab.bsky.social reporting spatial proteomics with DNA PAINT #SMLMS2025
We're excited that the study is now out in Nature Nanotechnology @natnano.nature.com www.nature.com/articles/s41...
New paper online:
Ångström-resolution imaging of cell-surface glycans.
The molecular organization of sugars in the native #glycocalyx has been resolved at 9 ångström using bioorthogonal metabolic labeling and #superresolution imaging of DNA barcodes.
#Glycotime
www.nature.com/articles/s41...
Congratulations to everyone involved: @ipachmayr.bsky.social, @lumasullo.bsky.social, @susannereinhardt.bsky.social, Jisoo Kwon, Ondřej Skořepa, Maite Llop, Sylvia Herter, Marina Bacac and Christian Klein. (6/6)
Read the full story here: www.nature.com/articles/s41...
The shift from Type II to Type I function reveals a structure–function continuum for anti-CD20 antibodies, showing that receptor arrangements dictate mechanism of action. RESI provides a platform for structure-guided antibody development, applicable far beyond CD20. (5/6)
We showed a direct link between CD20 oligomerization and function by investigating OBZ-based T-cell engagers (TCEs). An increased IgG flexibility in the 2+1 TCE format lead to increased CD20 tetramerization, without higher-order clustering, resulting in a reduction of direct cytotoxicity. (4/6)
