Information processing inside the central nervous system takes place on multiple scales in both 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.

Correlative two-photon and light sheet microscopy / Silvestri, L; Allegra Mascaro, A.L.; Costantini, I; Sacconi, L; Pavone, F.S.. - In: METHODS. - ISSN 1046-2023. - ELETTRONICO. - 66:(2014), pp. 268-272. [10.1016/j.ymeth.2013.06.013]

Correlative two-photon and light sheet microscopy

SILVESTRI, LUDOVICO;ALLEGRA MASCARO, ANNA LETIZIA;COSTANTINI, IRENE;SACCONI, LEONARDO;PAVONE, FRANCESCO SAVERIO
2014

Abstract

Information processing inside the central nervous system takes place on multiple scales in both 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.
2014
66
268
272
Silvestri, L; Allegra Mascaro, A.L.; Costantini, I; Sacconi, L; Pavone, F.S.
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/872318
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