Klimas Aleksandra, Gallagher Brendan, Zhao Yongxin
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania.
Curr Protoc Cytom. 2019 Dec;91(1):e67. doi: 10.1002/cpcy.67.
Optical imaging techniques are often used in neuroscience to understand brain function and discern disease pathogenesis. However, the optical diffraction limit precludes conventional optical imaging approaches from resolving nanoscopic structures with feature sizes smaller than 300 nm. Expansion microscopy (ExM) circumvents this limit by physically expanding preserved tissues embedded in a swellable hydrogel. Biomolecules of interest are covalently linked to a polymer matrix, which is then isotropically expanded at least 100-fold in size in pure water after mechanical homogenization of the tissue-gel. The sample can then be investigated with nanoscale precision using a conventional diffraction-limited microscope. The protocol described here is a variant of ExM that uses regents and equipment found in a typical biology laboratory and has been optimized for imaging proteins in expanded brain tissues. © 2019 by John Wiley & Sons, Inc. Basic Protocol: Expansion microscopy for intact brain tissue.
光学成像技术常用于神经科学领域，以了解脑功能并识别疾病发病机制。然而，光学衍射极限使得传统光学成像方法无法分辨特征尺寸小于300纳米的纳米级结构。扩展显微镜技术（ExM）通过物理方式扩展嵌入可膨胀水凝胶中的保存组织来克服这一限制。将感兴趣的生物分子共价连接到聚合物基质上，然后在对组织 - 凝胶进行机械匀浆后，使其在纯水中各向同性地膨胀至少100倍。然后可以使用传统的衍射极限显微镜以纳米级精度对样品进行研究。这里描述的方案是ExM的一种变体，它使用典型生物学实验室中常见的试剂和设备，并针对扩展脑组织中的蛋白质成像进行了优化。© 2019 John Wiley & Sons, Inc. 基本方案：完整脑组织的扩展显微镜技术。
