Department of Genetics and Genomics, Baylor College of Medicine;
Department of Genetics and Genomics, Baylor College of Medicine; Medical Scientist Training Program, Baylor College of Medicine.
J Vis Exp. 2021 Apr 20(170). doi: 10.3791/62532.
Brain activity, the electrochemical signals passed between neurons, is determined by the connectivity patterns of neuronal networks, and from the morphology of processes and substructures within these neurons. As such, much of what is known about brain function has arisen alongside developments in imaging technologies that allow further insight into how neurons are organized and connected in the brain. Improvements in tissue clearing have allowed for high-resolution imaging of thick brain slices, facilitating morphological reconstruction and analyses of neuronal substructures, such as dendritic arbors and spines. In tandem, advances in image processing software provide methods of quickly analyzing large imaging datasets. This work presents a relatively rapid method of processing, visualizing, and analyzing thick slices of labeled neural tissue at high-resolution using CLARITY tissue clearing, confocal microscopy, and image analysis. This protocol will facilitate efforts toward understanding the connectivity patterns and neuronal morphologies that characterize healthy brains, and the changes in these characteristics that arise in diseased brain states.
脑活动是神经元之间传递的电化学信号，由神经元网络的连接模式决定，并取决于这些神经元内的过程和亚结构的形态。因此，人们对大脑功能的了解在很大程度上是随着成像技术的发展而产生的，这些技术使我们能够进一步深入了解神经元在大脑中的组织和连接方式。组织透明化的改进使得对厚脑切片进行高分辨率成像成为可能，这有助于对神经元亚结构（如树突棘和树突）进行形态重建和分析。与此同时，图像处理软件的进步提供了快速分析大型成像数据集的方法。本工作提出了一种相对快速的方法，使用 CLARITY 组织透明化、共聚焦显微镜和图像分析来处理、可视化和分析高分辨率的标记神经组织的厚切片。该方案将有助于人们了解健康大脑的连接模式和神经元形态，以及在疾病状态下这些特征的变化。
