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

Year: 2014

Authors: Silvestri L., Allegra Mascaro A.L., Costantini I., Sacconi L., Pavone SF.

Autors Affiliation: European Laboratory for Non-linear Spectroscopy, University of Florence, Via Nello Carrara, 1, 50019 Sesto Fiorentino (FI), Italy; National Institute of Optics, National Research Council, Via Nello Carrara, 1, 50019 Sesto Fiorentino (FI), Italy; Department of Physics, University of Florence, Via G. Sansone, 1, 50019 Sesto Fiorentino (FI), Italy; International Center for Computational Neurophotonics (ICON Foundation), Via Nello Carrara, 1, 50019 Sesto Fiorentino (FI), Italy

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.

Conference title: Conference on Multiphoton Microscopy in the Biomedical Sciences XIV
Place: San Francisco, CA

KeyWords: Blood vessels; Brain mapping; Imaging techniques; Neuroimaging; Neurons; Photons, Brain imaging; Brain reconstruction; Correlative approaches; Correlative microscopies; Light-sheet microscopies; Spatio-temporal scale; Two-photon fluorescence imaging; Two-photon fluorescence microscopy, Brain
DOI: 10.1117/12.2037867

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