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

Summary: 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 reco...

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Main Authors: Keimpe Wierda, Hajnalka Nyitrai, Amélie Lejeune, Ine Vlaeminck, Elke Leysen, Tom Theys, Joris de Wit, Pierre Vanderhaeghen, Baptiste Libé-Philippot
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724004787
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author Keimpe Wierda
Hajnalka Nyitrai
Amélie Lejeune
Ine Vlaeminck
Elke Leysen
Tom Theys
Joris de Wit
Pierre Vanderhaeghen
Baptiste Libé-Philippot
author_facet Keimpe Wierda
Hajnalka Nyitrai
Amélie Lejeune
Ine Vlaeminck
Elke Leysen
Tom Theys
Joris de Wit
Pierre Vanderhaeghen
Baptiste Libé-Philippot
author_sort Keimpe Wierda
collection DOAJ
description Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
format Article
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institution Kabale University
issn 2666-1667
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publishDate 2024-12-01
publisher Elsevier
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spelling doaj-art-37763152edc449bb9bab7fe94e751da02024-11-20T05:07:46ZengElsevierSTAR Protocols2666-16672024-12-0154103313Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostainingKeimpe Wierda0Hajnalka Nyitrai1Amélie Lejeune2Ine Vlaeminck3Elke Leysen4Tom Theys5Joris de Wit6Pierre Vanderhaeghen7Baptiste Libé-Philippot8VIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; Electrophysiology Unit, VIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; Corresponding authorVIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; KUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, BelgiumVIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; KUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, BelgiumVIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; Electrophysiology Unit, VIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, BelgiumVIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; KUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, BelgiumKUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, Belgium; Research Group Experimental Neurosurgery and Neuroanatomy, KUL, 3000 Leuven, BelgiumVIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; KUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, Belgium; Corresponding authorVIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; KUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, Belgium; Corresponding authorVIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; KUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, Belgium; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724004787cell biologyneuroscienceprotein expression and purification
spellingShingle Keimpe Wierda
Hajnalka Nyitrai
Amélie Lejeune
Ine Vlaeminck
Elke Leysen
Tom Theys
Joris de Wit
Pierre Vanderhaeghen
Baptiste Libé-Philippot
Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostaining
STAR Protocols
cell biology
neuroscience
protein expression and purification
title Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostaining
title_full Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostaining
title_fullStr Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostaining
title_full_unstemmed Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostaining
title_short Protocol to process fresh human cerebral cortex biopsies for patch-clamp recording and immunostaining
title_sort protocol to process fresh human cerebral cortex biopsies for patch clamp recording and immunostaining
topic cell biology
neuroscience
protein expression and purification
url http://www.sciencedirect.com/science/article/pii/S2666166724004787
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