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用于膜片钳记录和免疫染色的新鲜人类大脑皮质活检组织处理方案。

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

作者信息

Wierda Keimpe, Nyitrai Hajnalka, Lejeune Amélie, Vlaeminck Ine, Leysen Elke, Theys Tom, de Wit Joris, Vanderhaeghen Pierre, Libé-Philippot Baptiste

机构信息

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

VIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium; KUL, Department of Neurosciences, Leuven Brain Institute, 3000 Leuven, Belgium.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103313. doi: 10.1016/j.xpro.2024.103313. Epub 2024 Sep 17.

Abstract

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..

摘要

大脑皮质活检能够对天然的发育中和成熟的人类脑组织进行研究。在此,我们展示了一种从手术室到实验室处理人类皮质活检样本的方案。我们描述了使用专门的化学溶液进行运输和切片以制备用于膜片钳记录的活急性切片的步骤。然后,我们解释了组织固定和事后免疫染色的程序,以便将生理特性与形态特征及蛋白质检测相关联。有关此方案使用和执行的完整详细信息,请参考利贝 - 菲利普ot等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f9c/11424940/2fee3dab3dae/fx1.jpg

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