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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.

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|November 28, 2025
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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.

Keywords:
NeuroscienceOrganoidsStem Cells

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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.