6533b828fe1ef96bd1288e38
RESEARCH PRODUCT
Fast whole-brain imaging of seizures in zebrafish larvae by two-photon light-sheet microscopy
Leonardo SacconiLudovico SilvestriNatascia TisoGiuseppe SancataldoLapo TurriniDuccio FanelliPietro RicciFrancesco S. PavoneGiacomo MazzamutoMarie-caroline MuellenbroichGiuseppe VitoFrancesco Vanzisubject
Materials scienceepilepsy zebrafish calcium imaging light sheet imaging two photon imagingbrain01 natural sciencesQuantitative Biology - Quantitative MethodsArticle010309 optics03 medical and health scienceszebrafish brain imaging microscopy two-photon light sheetTwo-photon excitation microscopyNeuroimaging0103 physical sciencesZebrafish larvaeQuantitative Methods (q-bio.QM)030304 developmental biologytwo-photon0303 health sciencesimaginglight sheetzebrafishAtomic and Molecular Physics and OpticsSettore FIS/07 - Fisica Applicata(Beni Culturali Ambientali Biol.e Medicin)3. Good healthFOS: Biological sciencesLight sheet fluorescence microscopyQuantitative Biology - Neurons and CognitionBiophysicsmicroscopyNeurons and Cognition (q-bio.NC)Biotechnologydescription
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).
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2022-03-01 |