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Multiplexed and high-throughput neuronal fluorescence imaging with diffusible probes

Author(s)
Guo, Syuan-Ming; Veneziano, Remi; Gordonov, Simon; Li, Li; Danielson, Eric W; Perez de Arce, Karen; Park, Demian; Kulesa, Anthony Benjamin; Wamhoff, Eike-Christian; Blainey, Paul C.; Blainey, Paul C; Boyden, Edward; Cottrell, Jeffrey R.; Bathe, Mark; ... Show more Show less
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Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/
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Abstract
Synapses contain hundreds of distinct proteins whose heterogeneous expression levels are determinants of synaptic plasticity and signal transmission relevant to a range of diseases. Here, we use diffusible nucleic acid imaging probes to profile neuronal synapses using multiplexed confocal and super-resolution microscopy. Confocal imaging is performed using high-affinity locked nucleic acid imaging probes that stably yet reversibly bind to oligonucleotides conjugated to antibodies and peptides. Super-resolution PAINT imaging of the same targets is performed using low-affinity DNA imaging probes to resolve nanometer-scale synaptic protein organization across nine distinct protein targets. Our approach enables the quantitative analysis of thousands of synapses in neuronal culture to identify putative synaptic sub-types and co-localization patterns from one dozen proteins. Application to characterize synaptic reorganization following neuronal activity blockade reveals coordinated upregulation of the post-synaptic proteins PSD-95, SHANK3 and Homer-1b/c, as well as increased correlation between synaptic markers in the active and synaptic vesicle zones.
Date issued
2019-09
URI
https://hdl.handle.net/1721.1/126016
Department
Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Media Laboratory; McGovern Institute for Brain Research at MIT; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Publisher
Springer Science and Business Media LLC
Citation
Guo, Syuang-Ming et al. "Multiplexed and high-throughput neuronal fluorescence imaging with diffusible probes." Nature Communications 10, 4377 (September 2019): 4377 © 2019 The Author(s)
Version: Final published version
ISSN
2041-1723

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