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J Neurosci. 2015 Apr 8;35(14):5792-807. doi: 10.1523/JNEUROSCI.4274-14.2015.

Mapping synapses by conjugate light-electron array tomography.

Author information

1
Department of Molecular and Cellular Physiology, Stanford School of Medicine, Stanford University, Stanford California 94305, Allen Institute for Brain Science, Seattle, Washington 98103 forrest.collman@gmail.com.
2
Department of Molecular and Cellular Physiology, Stanford School of Medicine, Stanford University, Stanford California 94305.
3
Department of Cell Biology and Physiology, University of North Carolina School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599.
4
Department of Cell Biology and Physiology, University of North Carolina School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599, Neuroscience Center, University of North Carolina, Chapel Hill, North Carolina 27599, and.
5
Department of Molecular and Cellular Physiology, Stanford School of Medicine, Stanford University, Stanford California 94305, Allen Institute for Brain Science, Seattle, Washington 98103.

Abstract

Synapses of the mammalian CNS are diverse in size, structure, molecular composition, and function. Synapses in their myriad variations are fundamental to neural circuit development, homeostasis, plasticity, and memory storage. Unfortunately, quantitative analysis and mapping of the brain's heterogeneous synapse populations has been limited by the lack of adequate single-synapse measurement methods. Electron microscopy (EM) is the definitive means to recognize and measure individual synaptic contacts, but EM has only limited abilities to measure the molecular composition of synapses. This report describes conjugate array tomography (AT), a volumetric imaging method that integrates immunofluorescence and EM imaging modalities in voxel-conjugate fashion. We illustrate the use of conjugate AT to advance the proteometric measurement of EM-validated single-synapse analysis in a study of mouse cortex.

KEYWORDS:

correlative microscopy; electron microscopy; immunofluorescence; synapses; synaptic diversity

Comment in

PMID:
25855189
PMCID:
PMC4388933
DOI:
10.1523/JNEUROSCI.4274-14.2015
[Indexed for MEDLINE]
Free PMC Article

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