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Pàgina inicial > Articles > Articles publicats > FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons |
Data: | 2015 |
Resum: | 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. |
Ajuts: | Ministerio de Economía y Competitividad SAF2013-42445R Ministerio de Economía y Competitividad BFU2008-03980 Ministerio de Economía y Competitividad SAF2013-2010-19930 Ministerio de Economía y Competitividad PIE13-00027 Ministerio de Economía y Competitividad BFU2012-34963 European Commission 604102 |
Drets: | Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. |
Llengua: | Anglès |
Document: | Article ; recerca ; Versió publicada |
Matèria: | Dendritic spines ; Synapses ; 3D-reconstruction ; Electron microscopy ; FIB/SEM ; Adult neurogenesis |
Publicat a: | Frontiers in Neuroanatomy, Vol. 9 (may 2015) , ISSN 1662-5129 |
17 p, 6.2 MB |