Scientific Results

Correlative two-photon and light sheet microscopy

Year: 2014

Authors: Silvestri L., Allegra Mascaro A. M., Costantini I., Sacconi L., Pavone F. S.

Autors Affiliation: European Laboratory for Non-Linear Spectroscopy, University of Florence, Italy; National Institute of Optics, National Research Council, Sesto Fiorentino, Italy; Department of Physics and Astronomy, University of Florence, Italy; International Center for Computational Neurophotonics (ICON Foundation), Sesto Fiorentino, Italy

Abstract: Information processing inside the central nervous system takes place on multiple scales in both space and time. A single imaging technique can reveal only a small part of this complex machinery. To obtain a more comprehensive view of brain functionality, complementary approaches should be combined into a correlative framework. Here, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, taking advantage of blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living thy1-GFP-M mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from the apical portion, the whole pyramidal neuron can then be segmented. The correlative approach presented here allows contextualizing within a three-dimensional anatomic framework the neurons whose dynamics have been observed with high detail in vivo. (C) 2013 Elsevier Inc. All rights reserved.

Journal/Review: METHODS

Volume: 66 (2)      Pages from: 268  to: 272

More Information: This work has received funding from LASERLAB-EUROPE (grant agreements nos. 228334 and 284464, EC\’s Seventh Framework Programme) and has been supported by Human Frontier Science Program research grant (RGP0027/2009), by the Italian Ministry for Education, University and Research in the framework of the Flagship Project NANOMAX, by Italian Ministry of Health in the framework of the \’Stem Cells Call for proposals\’. This work has been carried out in the framework of the research activities of ICON foundation supported by \”Ente Cassa di Risparmio di Firenze\”. This work is part of the research activities of the European Flasghip Human Brain Project.
KeyWords: Animal experiment; Animal tissue; Article; Brain blood vessel; Brain cortex; Confocal light sheet microscopy; Confocal microscopy; Dendritic spine; In vivo study; Mouse; Neurite; Neuroimaging; Nonhuman; Priority journal; Pyramidal nerve cell; Three dimensional imaging; Tissue fixation; Animal; Biosynthesis; Confocal microscopy; cytology; Dendrite; Fluorescence microscopy; Procedures; Somatosensory cortex; Transgenic mouse; Ultrastructure; Vascularization, Mus, green fluorescent protein, Animals; Dendrites; Green Fluorescent Proteins; Mice, Transgenic; Microscopy, Confocal; Microscopy, Fluorescence; Somatosensory Cortex
DOI: 10.1016/j.ymeth.2013.06.013

Citations: 19
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