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Proceedings Paper

Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy
Author(s): Ludovico Silvestri; Anna Letizia Allegra Mascaro; Irene Costantini; Leonardo Sacconi; Francesco S. Pavone
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Paper Abstract

One of the unique features of the brain is that its activity cannot be framed in a single spatio-temporal scale, but rather spans many orders of magnitude both in 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.

Paper Details

Date Published: 28 February 2014
PDF: 6 pages
Proc. SPIE 8948, Multiphoton Microscopy in the Biomedical Sciences XIV, 89480I (28 February 2014); doi: 10.1117/12.2037867
Show Author Affiliations
Ludovico Silvestri, European Lab. for Non-linear Spectroscopy (Italy)
Anna Letizia Allegra Mascaro, European Lab. for Non-linear Spectroscopy (Italy)
Irene Costantini, European Lab. for Non-linear Spectroscopy (Italy)
Leonardo Sacconi, National Institute of Optics, CNR (Italy)
European Lab. for Non-linear Spectroscopy (Italy)
Francesco S. Pavone, European Lab. for Non-linear Spectroscopy (Italy)
National Institute of Optics, CNR (Italy)
Univ. of Florence (Italy), and International Ctr. for Computational Neurophotonics (Italy)


Published in SPIE Proceedings Vol. 8948:
Multiphoton Microscopy in the Biomedical Sciences XIV
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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