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

Area of Science:

• Neuroscience
• Biotechnology
• Microscopy

Background:

• Brain organoids are revolutionizing neuroscience research by mimicking brain regions in vitro.
• Characterizing the electrical activity of brain organoids at single-neuron resolution in 3D remains a significant challenge.
• Existing methods are limited by the complexity of optical systems and biological samples.

Purpose of the Study:

• To introduce an accessible, minimalistic light-sheet microscope for 3D brain organoid imaging.
• To enable high-temporal-resolution, volumetric calcium imaging in brain organoids.
• To facilitate the study of neuronal network formation and connectivity in vitro.

Main Methods:

• Assembly of a minimalistic light-sheet microscope as an add-on to a standard inverted microscope.
• Utilizing a lightweight piezo stage for 5 Hz volumetric scanning.
• Development of a processing pipeline for true 3D neuronal trace segmentation.
• Imaging calcium activity in stem cell-derived neuron spheroids.

Main Results:

• Successful extraction of 4D calcium traces at high temporal resolution.
• Demonstration of volumetric scanning capabilities for comprehensive 3D imaging.
• Creation of a 3D connectivity map of a neuron spheroid.
• Achieved true 3D neuronal trace segmentation.

Conclusions:

• The developed light-sheet microscope provides an accessible platform for 3D brain organoid imaging.
• This technology empowers researchers to study neuronal network formation in vitro.
• Applications include fundamental neuroscience research and studies of neurodegeneration.