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Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostaining.

Keimpe Wierda1, Hajnalka Nyitrai2, Amélie Lejeune2

  • 1VIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; Electrophysiology Unit, VIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium.

STAR Protocols
|September 18, 2024
PubMed
Summary

This study details a protocol for processing human cerebral cortex biopsies. It enables acute slice preparation for electrophysiology and subsequent analysis of brain tissue properties.

Keywords:
cell biologyneuroscienceprotein expression and purification

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Area of Science:

  • Neuroscience
  • Human Brain Tissue Research
  • Biopsy Processing

Background:

  • Investigating human brain tissue is crucial for understanding development and disease.
  • Access to native, viable human brain tissue is limited.
  • Current methods may not preserve tissue integrity for detailed analysis.

Purpose of the Study:

  • To present a comprehensive protocol for processing human cerebral cortex biopsies.
  • To enable the preparation of viable acute slices for electrophysiological recordings.
  • To facilitate correlation of physiological data with morphological and molecular features.

Main Methods:

  • Describing a step-by-step protocol from surgical theater to laboratory.
  • Utilizing specialized chemical solutions for tissue transport and sectioning.
  • Implementing tissue fixation and post hoc immunostaining techniques.

Main Results:

  • Successful preparation of viable acute human cerebral cortex slices.
  • Demonstration of patch-clamp recording feasibility on processed tissue.
  • Establishment of methods to link electrophysiological properties with cellular morphology and protein expression.

Conclusions:

  • The presented protocol provides a reliable method for studying human brain tissue.
  • This approach supports integrated physiological, morphological, and molecular investigations.
  • The protocol enhances research into developing and mature human brain function and pathology.