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28 February 2014 Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy
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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, Anna Letizia Allegra Mascaro, Irene Costantini, Leonardo Sacconi, and 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);

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