Spatial proteomics in neurons at single-protein resolution

2024/03/01 by Eduard M. Unterauer, Sayedali Shetab Boushehri, Kristina Jevdokimenko +11 · 1 voice · 150 citations
Biochemistry, Genetics and Molecular Biology · #Advanced Fluorescence Microscopy Techniques #Artificial intelligence #Biology #Cell Image Analysis Techniques #Computational biology #Computer science #Gene #Genetics #Multiplexing #Proteomics #Resolution (logic) #Single-cell and spatial transcriptomics #Workflow

paper · pdf · doi:10.1016/j.cell.2024.02.045

published in Cell 187(7), 1785-1800.e16 (Cell Press)

openalex publication_date 2024/03/01 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/06

Abstract

To understand biological processes, it is necessary to reveal the molecular heterogeneity of cells by gaining access to the location and interaction of all biomolecules. Significant advances were achieved by super-resolution microscopy, but such methods are still far from reaching the multiplexing capacity of proteomics. Here, we introduce secondary label-based unlimited multiplexed DNA-PAINT (SUM-PAINT), a high-throughput imaging method that is capable of achieving virtually unlimited multiplexing at better than 15 nm resolution. Using SUM-PAINT, we generated 30-plex single-molecule resolved datasets in neurons and adapted omics-inspired analysis for data exploration. This allowed us to reveal the complexity of synaptic heterogeneity, leading to the discovery of a distinct synapse type. We not only provide a resource for researchers, but also an integrated acquisition and analysis workflow for comprehensive spatial proteomics at single-protein resolution.

Citations

• On a Test of Whether one of Two Random Variables is Stochastically Larger than the Other

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Discussions

• It's advanced talks day at #FOMISUR! Starting with @lumasullo.bsky.social demonstrating 30-color SUM-PAINT in neurons (doi.org/10.1016/j.ce...) as well as angstrom-resolution RESI (doi.org/10.1038/s41 [bsky, 24 points, 1 comments]

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