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. 2024 Mar 28;187(7):1785-1800.e16.
doi: 10.1016/j.cell.2024.02.045.

Spatial proteomics in neurons at single-protein resolution

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Free article

Spatial proteomics in neurons at single-protein resolution

Eduard M Unterauer et al. Cell. .
Free article

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.

Keywords: DNA-PAINT; excitatory synapses; inhibitory synapses; multiplexing; neuron atlas; neuron imaging; proteomics; spatial-omics; super-resolution microscopy; synapse; synapse diversity; synaptic proteins.

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Conflict of interest statement

Declaration of interests E.M.U., M.G. and R.J. have filed a patent on the method published in this paper. F.O. is a shareholder of NanoTag Biotechnologies GmbH. The other authors declare no competing financial interests.

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