Fast whole-brain imaging of seizures in zebrafish larvae by two-photon light-sheet microscopy

Year: 2022

Authors: De Vito G., Turrini L., Müllenbroich C., Ricci P., Sancataldo G., Mazzamuto G., Tiso N., Sacconi L., Fanelli D., Silvestri L., Vanzi F., Pavone FS.

Autors Affiliation: University of Florence, Department of Neuroscience, Psychology, Drug Research and Child Health, Viale Pieraccini 6, Florence, Italy, 50139, Italy European Laboratory for Non-Linear Spectroscopy, Via Nello Carrara 1, Sesto Fiorentino 50019, Italy University of Florence, Department of Physics and Astronomy, Via Sansone 1, Sesto Fiorentino 50019, Italy School of Physics and Astronomy, Kelvin Building, University of Glasgow, G12 8QQ, Glasgow, UK National Institute of Optics, National Research Council, Via Nello Carrara 1, Sesto Fiorentino 50019, Italy University of Padova, Department of Biology, Via U. Bassi 58/B, Padova 35131, Italy University of Florence, Department of Biology, Via Madonna del Piano 6, Sesto Fiorentino 50019, Italy

Abstract: Light-sheet fluorescence microscopy (LSFM) enables real-time whole-brain functional imaging in zebrafish larvae. Conventional one-photon LSFM can however induce undesirable visual stimulation due to the use of visible excitation light. The use of two-photon (2P) excitation, employing near-infrared invisible light, provides unbiased investigation of neuronal circuit dynamics. However, due to the low efficiency of the 2P absorption process, the imaging speed of this technique is typically limited by the signal-to-noise-ratio. Here, we describe a 2P LSFM setup designed for non-invasive imaging that enables quintuplicating state-of-the-art volumetric acquisition rate of the larval zebrafish brain (5 Hz) while keeping low the laser intensity on the specimen. We applied our system to the study of pharmacologically-induced acute seizures, characterizing the spatial-temporal dynamics of pathological activity and describing for the first time the appearance of caudo-rostral ictal waves (CRIWs).


Volume: 13      Pages from: 1516  to: 1536

KeyWords: two-photon, light-sheet microscopy, zebrafish, seizure, brain imaging
DOI: 10.1364/BOE.434146

Citations: 11
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