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bioRxiv · 10.1101/2023.05.24.542114

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

Chai, Y.·Qi, K.·Wu, Y.·Li, D.·Tan, G.·Guo, Y.·Chu, J.·Mu, Y.·Shen, C.·Wen, Q.
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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 freely swimming 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 demonstrating that selective light activation of the midbrain regions in larval zebrafish could reliably trigger biased turning behavior and changes of brain-wide neural activity, we present a valuable tool for investigating the causal relationship between distributed neural circuit dynamics and naturalistic behavior. HighlightsO_LIWe develop an all-optical technique that enables simultaneous whole brain imaging and optogenetic manipulation of selective brain regions in freely behaving larval zebrafish. C_LIO_LIA combination of a genetically encoded calcium indicator and a long Stokes-shift red fluorescence protein, together with the adaptive filter algorithm, enables us to reliably distinguish calcium activity from motion-induced signal contamination. C_LIO_LIRapid 3D image reconstruction and registration enables real-time targeted optogenetic stimulation of distinct brain regions in a freely swimming larval zebrafish. C_LI

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Chai, Y., Qi, K., Wu, Y., Li, D., Tan, G., Guo, Y., Chu, J., Mu, Y., Shen, C., Wen, Q.. 2023-05-24. All-optical interrogation of brain-wide activity in freely swimming larval zebrafish. https://doi.org/10.1101/2023.05.24.542114

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