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Title: FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
Author: Bosch, Carles
Martínez García, Albert
Masachs Janoher, Núria
Teixeira, Catia Marlene
Fernaud, Isabel
Ulloa Darquea, Fausto Alexander
Pérez Martínez, Esther
Lois, Carlos
Comella i Carnicé, Joan Xavier, 1963-
DeFelipe, Javier
Merchán Pérez, Angel
Soriano García, Eduardo
Keywords: Neurones
Issue Date: 21-May-2015
Publisher: Frontiers Media
Abstract: The fine analysis of synaptic contacts is usually performed using transmission electron microscopy (TEM) and its combination with neuronal labeling techniques. However, the complex 3D architecture of neuronal samples calls for their reconstruction from serial sections. Here we show that focused ion beam/scanning electron microscopy (FIB/SEM) allows efficient, complete, and automatic 3D reconstruction of identified dendrites, including their spines and synapses, from GFP/DAB-labeled neurons, with a resolution comparable to that of TEM. We applied this technology to analyze the synaptogenesis of labeled adult-generated granule cells (GCs) in mice. 3D reconstruction of dendritic spines in GCs aged 3-4 and 8-9 weeks revealed two different stages of dendritic spine development and unexpected features of synapse formation, including vacant and branched dendritic spines and presynaptic terminals establishing synapses with up to 10 dendritic spines. Given the reliability, efficiency, and high resolution of FIB/SEM technology and the wide use of DAB in conventional EM, we consider FIB/SEM fundamental for the detailed characterization of identified synaptic contacts in neurons in a high-throughput manner.
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It is part of: Frontiers In Neuroanatomy, 2015, vol. 9, p. 60
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ISSN: 1662-5129
Appears in Collections:Articles publicats en revistes (Biologia Cel·lular, Fisiologia i Immunologia)

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