A minimal-complexity light-sheet microscope maps network activity in 3D neuronal systems

Wysmolek, Paulina M.; Kiessler, Filippo D.; Salbaum, Katja A.; Shelton, Elijah R.; Sonntag, Selina M. und Serwane, Friedhelm (28. November 2022): A minimal-complexity light-sheet microscope maps network activity in 3D neuronal systems. In: Scientific Reports, Bd. 12, Nr. 1

Abstract

In vitro systems mimicking brain regions, brain organoids, are revolutionizing the neuroscience field. However, characterization of their electrical activity has remained a challenge as it requires readout at millisecond timescale in 3D at single-neuron resolution. While custom-built microscopes used with genetically encoded sensors are now opening this door, a full 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 an accessible minimalistic light-sheet microscope to the neuroscience community. Designed as an add-on to a standard inverted microscope it can be assembled within one day. In contrast to existing simplistic setups, our platform is suited to record volumetric calcium traces. We successfully extracted 4D calcium traces at high temporal resolution by using a lightweight piezo stage to allow for 5 Hz volumetric scanning combined with a processing pipeline for true 3D neuronal trace segmentation. As a proof of principle, we created a 3D connectivity map of a stem cell derived neuron spheroid by imaging its activity. Our fast, low complexity setup empowers researchers to study the formation of neuronal networks in vitro for fundamental and neurodegeneration research.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Physik ; Medizin > Munich Cluster for Systems Neurology (SyNergy)
• Themengebiete: 500 Naturwissenschaften und Mathematik > 530 Physik
• ISSN: 2045-2322
• Sprache: Englisch
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 491502892
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 390857198