All-optical interrogation of brain-wide activity in freely swimming larval zebrafish.

ABSTRACT: We introduce an all-optical technique that enables volumetric imaging of brain-wide calcium activity and targeted optogenetic stimulation of specific brain regions in unrestrained larval zebrafish. The system consists of three main components: a 3D tracking module, a dual-color fluorescence imaging module, and a real-time activity manipulation module. Our approach uses a sensitive genetically encoded calcium indicator in combination with a long Stokes shift red fluorescence protein as a reference channel, allowing the extraction of Ca²⁺ activity from signals contaminated by motion artifacts. The method also incorporates rapid 3D image reconstruction and registration, facilitating real-time selective optogenetic stimulation of different regions of the brain. By demonstrati

SUBMITTER: Chai Y 

PROVIDER: S-EPMC10776927 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature