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Spatial proteomics in neurons at single-protein resolution.
Cell 187, 1785-1800.e16 (2024)
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.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Dna-paint ; Excitatory Synapses ; Inhibitory Synapses ; Multiplexing ; Neuron Atlas ; Neuron Imaging ; Proteomics ; Spatial-omics ; Super-resolution Microscopy ; Synapse ; Synapse Diversity ; Synaptic Proteins; Superresolution Microscopy; Dna-paint; Release; Binding
ISSN (print) / ISBN
0092-8674
e-ISSN
1097-4172
Journal
Cell
Quellenangaben
Volume: 187,
Issue: 7,
Pages: 1785-1800.e16
Publisher
Cell Press
Publishing Place
Cambridge, Mass.
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of AI for Health (AIH)
Grants
ERC
BMBF
Max Planck Foundation
Max Planck Society
QBM graduate school
IMPRS-ML graduate school
European Union
Deutsche Forschungsgemeinschaft (DFG)
Schram Stiftung
Hightech Agenda Bayern
Collaborative Research Center 1286 on Quantitative Synaptologie Gottingen, Germany
F. Hoffmann-La Roche
Helmholtz Association
European Research Council
BMBF
Max Planck Foundation
Max Planck Society
QBM graduate school
IMPRS-ML graduate school
European Union
Deutsche Forschungsgemeinschaft (DFG)
Schram Stiftung
Hightech Agenda Bayern
Collaborative Research Center 1286 on Quantitative Synaptologie Gottingen, Germany
F. Hoffmann-La Roche
Helmholtz Association
European Research Council