Share Email Print
cover

Proceedings Paper

Two-photon high-speed light-sheet volumetric imaging of brain activity during sleep in zebrafish larvae
Author(s): Giuseppe de Vito; Chiara Fornetto; Pietro Ricci; Caroline Müllenbroich; Giuseppe Sancataldo; Lapo Turrini; Giacomo Mazzamuto; Natascia Tiso; Leonardo Sacconi; Duccio Fanelli; Ludovico Silvestri; Francesco Vanzi; Francesco Saverio Pavone
Format Member Price Non-Member Price
PDF $17.00 $21.00

Paper Abstract

Although it is well known that zebrafish display the behavioural signature of sleep, the neuronal correlates of this state are not yet completely understood, due to the complexity of the measurements required. For example, when performed with visible excitation light, functional imaging can disrupt the day/night cycle due to the induced visual stimulation. To address this issue, we developed a custom-made two-photon light-sheet microscope optimized for high-speed volumetric imaging. By employing infra-red light (not visible to the larva) for excitation, we are able to record wholebrain neuronal activity with high temporal- and spatial-resolution without affecting the sleep state. In two-photon light-sheet microscopy the maximum achievable frame rate is limited by the signal-to-noise ratio. To maximize this parameter, we optimized our setup for high peak power of excitation light, while finely controlling its polarisation, and we implemented remote scanning of the focal plane to record without disturbing the sample. Using this setup, as a preliminary result, we characterized the intensity spectra of neuronal calcium traces of 4 days post fertilisation larvae during the day/night phases. We aim to extend these results to multiple brain regions and frequency bands.

Paper Details

Date Published: 21 February 2020
PDF: 4 pages
Proc. SPIE 11226, Neural Imaging and Sensing 2020, 1122604 (21 February 2020); doi: 10.1117/12.2542285
Show Author Affiliations
Giuseppe de Vito, Univ. degli Studi di Firenze (Italy)
LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Chiara Fornetto, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Pietro Ricci, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Caroline Müllenbroich, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Univ. of Glasgow (United Kingdom)
Istituto Nazionale di Ottica, Consiglio Nazionale delle Ricerche (Italy)
Giuseppe Sancataldo, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Univ. degli Studi di Firenze (Italy)
Lapo Turrini, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Univ. degli Studi di Firenze (Italy)
Giacomo Mazzamuto, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Istituto Nazionale di Ottica, Consiglio Nazionale delle Ricerche (Italy)
Natascia Tiso, Univ. degli Studi di Padova (Italy)
Leonardo Sacconi, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Istituto Nazionale di Ottica, Consiglio Nazionale delle Ricerche (Italy)
Duccio Fanelli, Univ. degli Studi di Firenze (Italy)
Ludovico Silvestri, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Istituto Nazionale di Ottica, Consiglio Nazionale delle Ricerche (Italy)
Univ. degli Studi di Firenze (Italy)
Francesco Vanzi, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Univ. degli Studi di Firenze (Italy)
Francesco Saverio Pavone, LENS - Lab. Europeo di Spettroscopie Non-Lineari (Italy)
Istituto Nazionale di Ottica, Consiglio Nazionale delle Ricerche (Italy)
Univ. degli Studi di Firenze (Italy)


Published in SPIE Proceedings Vol. 11226:
Neural Imaging and Sensing 2020
Qingming Luo; Jun Ding; Ling Fu, Editor(s)

© SPIE. Terms of Use
Back to Top
PREMIUM CONTENT
Sign in to read the full article
Create a free SPIE account to get access to
premium articles and original research
Forgot your username?
close_icon_gray