Measuring Electrophysiological Activity in Acute Brain Slices, Spheroids, and Organoids Using 3D High-Density...

Area of Science:

• Neuroscience
• Biotechnology
• Electrophysiology

Background:

• Three-dimensional (3D) brain models offer superior recapitulation of in vivo neural network properties compared to 2D models.
• Standard planar multielectrode arrays (MEAs) exhibit limitations with 3D models, including poor adhesion, insufficient sensing elements, and compromised tissue vitality.
• 3D high-density MEAs (3D HD-MEAs) represent an advancement, improving sensing capabilities and supporting 3D model viability.

Purpose of the Study:

• To present an optimized protocol for neural network activity recordings in various 3D brain models using 3D HD-MEAs.
• To provide researchers with a comprehensive guide for acquiring high-quality electrophysiological data from acute slices, brain spheroids, and organoids.
• To detail critical steps, offer practical tips, and suggest troubleshooting for reproducible functional recordings.

Main Methods:

• Preparation of viable acute brain slices from specific mouse brain regions (cerebellum, cortico-hippocampal circuit, prefrontal cortex).
• Establishment of efficient coupling between brain slices and 3D HD-MEA chips.
• Standardized procedures for obtaining and recording from human and animal brain spheroids and neural organoids.
• Utilizing BrainWave6 software for data acquisition, signal detection, and electrophysiological analysis.

Main Results:

• Demonstrated successful implementation of 3D HD-MEAs for recording neural activity from diverse 3D brain models.
• Validated an optimized protocol ensuring efficient coupling, enhanced tissue vitality, and high-quality signal retrieval.
• Provided specific adjustments for different brain areas and sample types, highlighting crucial steps and troubleshooting strategies.

Conclusions:

• The presented protocol optimizes the use of 3D HD-MEAs for electrophysiological recordings from 3D brain models.
• This guide facilitates efficient and reproducible functional recordings, maximizing the potential of advanced 3D neural models.
• The optimized protocol serves as a valuable resource for researchers studying brain function and pathology in vitro.