PT  - JOURNAL ARTICLE
AU  - Paulina M. Wysmolek
AU  - Filippo D. Kiessler
AU  - Katja A. Salbaum
AU  - Elijah R. Shelton
AU  - Selina Sonntag
AU  - Friedhelm Serwane
TI  - Light-sheet microscopy of network activity in 3D neuronal systems
AID  - 10.1101/2022.06.20.496852
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.06.20.496852
4099  - http://biorxiv.org/content/early/2022/06/21/2022.06.20.496852.short
4100  - http://biorxiv.org/content/early/2022/06/21/2022.06.20.496852.full
AB  - In vitro systems mimicking brain regions, such as brain organoids, are revolutionizing the field of neuroscience. However, characterization of their electrical activity has remained a challenge as this requires electrophysiological readout at millisecond timescale in 3D at single-neuron resolution. While custom-built microscopes used with genetically encoded sensors start to grant this access, a thorough 3D characterization of organoid neural activity has not been performed yet, limited by the combined complexity of the optical and the biological system. Here, we introduce a simple microscope for volumetric characterization of network activity with single-neuron resolution. To provide a versatile, accessible platform to the neuroscience community, we designed a minimalistic light-sheet microscope tailored to computational neuroscience tools to extract calcium traces. As a proof of principle, we created a 3D connectivity map of the neuronal network by imaging its spontaneous activity. High performance, low complexity setups such as ours will empower researchers to study the formation of neuronal networks in vitro for fundamental and neurodegeneration research.Competing Interest StatementThe authors have declared no competing interest.