Protocol for recording of neural activity in 3D midbrain organoids using a multiple-electrode array.
Asiye Malkoç1, Diana Riekschnitz1, Marcelo Rosato Siri1
1Eurac Research, Institute for Biomedicine, A.-Volta-Straße 21/Via A.-Volta 21, 39100 Bozen/Bolzano, Italy.
STAR Protocols
|November 28, 2025
Summary
This study details a new protocol for electrophysiological recording from human brain organoids. The method uses multi-electrode array plates for reliable neural activity analysis in neurodevelopment research.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Biotechnology
Background:
- Human organoids offer a model for studying neurodevelopment and diseases.
- Electrophysiological recording is crucial for understanding neuronal function.
- Existing methods may not be optimal for 3D organoid cultures.
Purpose of the Study:
- To present a protocol for electrophysiological recording from human 3D organoids.
- To adapt 2D culture substrates for 3D organoid electrophysiology.
- To enable reliable and reproducible neural activity measurements.
Main Methods:
- Generation of midbrain 3D organoids from human induced pluripotent stem cells (hiPSCs).
- Utilization of 6-well multi-electrode array (MEA) plates for organoid culture and recording.
- Application of the neural module for Axion Maestro system for data acquisition.
Main Results:
- Successful electrophysiological recording from human 3D organoids.
- Demonstration of high reliability and reproducibility in neural recordings.
- Validation of the protocol for capturing neuronal activity.
Conclusions:
- The presented protocol enables robust electrophysiological analysis of human 3D organoids.
- This method advances research in human neurodevelopment and disease modeling.
- Opens new avenues for studying complex neural circuits in vitro.
