Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy

Abstract Optical recordings of neuronal activity at cellular resolution represent an invaluable tool to investigate brain mechanisms. Zebrafish larvae is one of the few model organisms where, using fluorescence-based reporters of the cell activity, it is possible to optically reconstruct the neurona...

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Autores principales:	Matteo Bruzzone, Enrico Chiarello, Marco Albanesi, Maria Elena Miletto Petrazzini, Aram Megighian, Claudia Lodovichi, Marco dal Maschio
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/95c312c8227c47b0b74ed7d82899e76b
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spelling	oai:doaj.org-article:95c312c8227c47b0b74ed7d82899e76b2021-12-02T15:00:51ZWhole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy10.1038/s41598-021-90335-y2045-2322https://doaj.org/article/95c312c8227c47b0b74ed7d82899e76b2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90335-yhttps://doaj.org/toc/2045-2322Abstract Optical recordings of neuronal activity at cellular resolution represent an invaluable tool to investigate brain mechanisms. Zebrafish larvae is one of the few model organisms where, using fluorescence-based reporters of the cell activity, it is possible to optically reconstruct the neuronal dynamics across the whole brain. Typically, leveraging the reduced light scattering, methods like lightsheet, structured illumination, and light-field microscopy use spatially extended excitation profiles to detect in parallel activity signals from multiple cells. Here, we present an alternative design for whole brain imaging based on sequential 3D point-scanning excitation. Our approach relies on a multiphoton microscope integrating an electrically tunable lens. We first apply our approach, adopting the GCaMP6s activity reporter, to detect functional responses from retinal ganglion cells (RGC) arborization fields at different depths within the zebrafish larva midbrain. Then, in larvae expressing a nuclear localized GCaMP6s, we recorded whole brain activity with cellular resolution. Adopting a semi-automatic cell segmentation, this allowed reconstructing the activity from up to 52,000 individual neurons across the brain. In conclusion, this design can easily retrofit existing imaging systems and represents a compact, versatile and reliable tool to investigate neuronal activity across the larva brain at high resolution.Matteo BruzzoneEnrico ChiarelloMarco AlbanesiMaria Elena Miletto PetrazziniAram MegighianClaudia LodovichiMarco dal MaschioNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Matteo Bruzzone Enrico Chiarello Marco Albanesi Maria Elena Miletto Petrazzini Aram Megighian Claudia Lodovichi Marco dal Maschio Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy
description	Abstract Optical recordings of neuronal activity at cellular resolution represent an invaluable tool to investigate brain mechanisms. Zebrafish larvae is one of the few model organisms where, using fluorescence-based reporters of the cell activity, it is possible to optically reconstruct the neuronal dynamics across the whole brain. Typically, leveraging the reduced light scattering, methods like lightsheet, structured illumination, and light-field microscopy use spatially extended excitation profiles to detect in parallel activity signals from multiple cells. Here, we present an alternative design for whole brain imaging based on sequential 3D point-scanning excitation. Our approach relies on a multiphoton microscope integrating an electrically tunable lens. We first apply our approach, adopting the GCaMP6s activity reporter, to detect functional responses from retinal ganglion cells (RGC) arborization fields at different depths within the zebrafish larva midbrain. Then, in larvae expressing a nuclear localized GCaMP6s, we recorded whole brain activity with cellular resolution. Adopting a semi-automatic cell segmentation, this allowed reconstructing the activity from up to 52,000 individual neurons across the brain. In conclusion, this design can easily retrofit existing imaging systems and represents a compact, versatile and reliable tool to investigate neuronal activity across the larva brain at high resolution.
format	article
author	Matteo Bruzzone Enrico Chiarello Marco Albanesi Maria Elena Miletto Petrazzini Aram Megighian Claudia Lodovichi Marco dal Maschio
author_facet	Matteo Bruzzone Enrico Chiarello Marco Albanesi Maria Elena Miletto Petrazzini Aram Megighian Claudia Lodovichi Marco dal Maschio
author_sort	Matteo Bruzzone
title	Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy
title_short	Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy
title_full	Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy
title_fullStr	Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy
title_full_unstemmed	Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy
title_sort	whole brain functional recordings at cellular resolution in zebrafish larvae with 3d scanning multiphoton microscopy
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/95c312c8227c47b0b74ed7d82899e76b
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Whole brain functional recordings at cellular resolution in zebrafish larvae with 3D scanning multiphoton microscopy

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