Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy

Ludovico Silvestri; Anna Letizia Allegra Mascaro; Irene Costantini; Leonardo Sacconi; Francesco S. Pavone

doi:10.1117/12.2037867

28 February 2014 Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy

Ludovico Silvestri, Anna Letizia Allegra Mascaro, Irene Costantini, Leonardo Sacconi, Francesco S. Pavone

Author Affiliations +

Proceedings Volume 8948, Multiphoton Microscopy in the Biomedical Sciences XIV; 89480I (2014) https://doi.org/10.1117/12.2037867
Event: SPIE BiOS, 2014, San Francisco, California, United States

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.

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Ludovico Silvestri, Ludovico Silvestri, Anna Letizia Allegra Mascaro, Anna Letizia Allegra Mascaro, Irene Costantini, Irene Costantini, Leonardo Sacconi, Leonardo Sacconi, Francesco S. Pavone, Francesco S. Pavone, "Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy", Proc. SPIE 8948, Multiphoton Microscopy in the Biomedical Sciences XIV, 89480I (28 February 2014); doi: 10.1117/12.2037867; https://doi.org/10.1117/12.2037867

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