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Proceedings Paper

Neural plasticity explored by correlative two-photon and electron/SPIM microscopy
Author(s): A. L. Allegra Mascaro; L. Silvestri; I. Costantini; L. Sacconi; B. Maco; G. W. Knott; F. S. Pavone
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Paper Abstract

Plasticity of the central nervous system is a complex process which involves the remodeling of neuronal processes and synaptic contacts. However, a single imaging technique can reveal only a small part of this complex machinery. To obtain a more complete view, complementary approaches should be combined. Two-photon fluorescence microscopy, combined with multi-photon laser nanosurgery, allow following the real-time dynamics of single neuronal processes in the cerebral cortex of living mice. The structural rearrangement elicited by this highly confined paradigm of injury can be imaged in vivo first, and then the same neuron could be retrieved ex-vivo and characterized in terms of ultrastructural features of the damaged neuronal branch by means of electron microscopy. Afterwards, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, based on the use of major blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from its apical portion, the whole pyramidal neuron can then be segmented and located in the correct cortical layer. With the correlative approach presented here, researchers will be able to place in a three-dimensional anatomic context the neurons whose dynamics have been observed with high detail in vivo.

Paper Details

Date Published: 17 June 2013
PDF: 6 pages
Proc. SPIE 8797, Advanced Microscopy Techniques III, 87970G (17 June 2013); doi: 10.1117/12.2032505
Show Author Affiliations
A. L. Allegra Mascaro, LENS, European Lab. for Non-linear Spectroscopy (Italy)
L. Silvestri, LENS, European Lab. for Non-linear Spectroscopy (Italy)
I. Costantini, LENS, European Lab. for Non-linear Spectroscopy (Italy)
L. Sacconi, LENS, European Lab. for Non-linear Spectroscopy (Italy)
International Ctr. Of Computational Neurophotonics (Italy)
B. Maco, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
G. W. Knott, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
F. S. Pavone, LENS, European Lab. for Non-linear Spectroscopy (Italy)
International Ctr. Of Computational Neurophotonics (Italy)
Univ. of Florence (Italy)


Published in SPIE Proceedings Vol. 8797:
Advanced Microscopy Techniques III
Emmanuel Beaurepaire; Peter T. C. So, Editor(s)

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