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

Résumé fourni par la source

Cerebral cortex biopsies enable the investigation of native developing and mature human brain tissue. Here, we present a protocol to process human cortical biopsies from the surgical theater to the laboratory. We describe steps for the preparation of viable acute slices for patch-clamp recording using dedicated chemical solutions for transport and sectioning. We then explain procedures for tissue fixation and post hoc immunostaining to correlate physiological properties to morphological features and protein detection. For complete details on the use and execution of this protocol, please refer to Libé-Philippot et al. 1 • Preparation of the biopsy transport from the surgical theater to the laboratory • Sectioning of the cortical biopsy to perform electrophysiological recordings • Post hoc immunostaining of the patched neurons to detect proteins herein • TSA fluorescence amplification to immunodetect proteins produced at low levels Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Cerebral cortex biopsies enable the investigation of native developing and mature human brain tissue. Here, we present a protocol to process human cortical biopsies from the surgical theater to the laboratory. We describe steps for the preparation of viable acute slices for patch-clamp recording using dedicated chemical solutions for transport and sectioning. We then explain procedures for tissue fixation and post hoc immunostaining to correlate physiological properties to morphological features and protein detection.

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