Two-photon high-speed light-sheet volumetric imaging of brain activity during sleep in zebrafish larvae

Giuseppe Sancataldo, Francesco Saverio Pavone, Giacomo Mazzamuto, Giuseppe De Vito, Ludovico Silvestri, Giuseppe Sancataldo, Lapo Turrini, Caroline Müllenbroich, Chiara Fornetto, Pietro Ricci, Leonardo Sacconi, Natascia Tiso, Duccio Fanelli, Francesco Vanzi

Risultato della ricerca: Conference articlepeer review

4 Citazioni (Scopus)

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.
Lingua originaleEnglish
pagine (da-a)3-
Numero di pagine4
RivistaPROGRESS IN BIOMEDICAL OPTICS AND IMAGING
Volume11226
Stato di pubblicazionePublished - 2020

All Science Journal Classification (ASJC) codes

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  • ???subjectarea.asjc.2500.2502???
  • ???subjectarea.asjc.3100.3107???
  • ???subjectarea.asjc.2700.2741???

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