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Correlative in vivo 2 photon and focused ion beam scanning electron microscopy of cortical neurons.

Type of publication Peer-reviewed
Publikationsform Original article (peer-reviewed)
Publication date 2013
Author Maco B, Holtmaat A, Cantoni M, Kreshuk A, Straehle CN, Hamprecht FA, Knott GW,
Project Long-term potentiation and the modification of synaptic structures in vivo
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Original article (peer-reviewed)

Journal PLoS One
Volume (Issue) 8(2)
Page(s) e57405
Title of proceedings PLoS One
DOI doi: 10.1371/journal.pone.0057405

Abstract

Correlating in vivo imaging of neurons and their synaptic connections with electron microscopy combines dynamic and ultrastructural information. Here we describe a semi-automated technique whereby volumes of brain tissue containing axons and dendrites, previously studied in vivo, are subsequently imaged in three dimensions with focused ion beam scanning electron microcopy. These neurites are then identified and reconstructed automatically from the image series using the latest segmentation algorithms. The fast and reliable imaging and reconstruction technique avoids any specific labeling to identify the features of interest in the electron microscope, and optimises their preservation and staining for 3D analysis.
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