Sensory Circuits and Neurotechnology Lab., The Francis Crick Institute, London, UK.
Department of Neuroscience, Physiology and Pharmacology, University College, London, UK.
Nat Commun. 2022 May 25;13(1):2923. doi: 10.1038/s41467-022-30199-6.
Understanding the function of biological tissues requires a coordinated study of physiology and structure, exploring volumes that contain complete functional units at a detail that resolves the relevant features. Here, we introduce an approach to address this challenge: Mouse brain tissue sections containing a region where function was recorded using in vivo 2-photon calcium imaging were stained, dehydrated, resin-embedded and imaged with synchrotron X-ray computed tomography with propagation-based phase contrast (SXRT). SXRT provided context at subcellular detail, and could be followed by targeted acquisition of multiple volumes using serial block-face electron microscopy (SBEM). In the olfactory bulb, combining SXRT and SBEM enabled disambiguation of in vivo-assigned regions of interest. In the hippocampus, we found that superficial pyramidal neurons in CA1a displayed a larger density of spine apparati than deeper ones. Altogether, this approach can enable a functional and structural investigation of subcellular features in the context of cells and tissues.
理解生物组织的功能需要对生理学和结构进行协调研究，探索包含完整功能单元的体积，其细节可解析相关特征。在这里，我们介绍了一种解决这一挑战的方法：使用体内双光子钙成像记录功能的区域的小鼠脑组织切片经过染色、脱水、树脂包埋，并用基于传播的相位对比同步辐射 X 射线计算机断层扫描（SXRT）进行成像。SXRT 以亚细胞细节提供了背景信息，并且可以通过使用连续块面电子显微镜（SBEM）进行靶向采集多个体积来跟进。在嗅球中，结合使用 SXRT 和 SBEM 可以对体内分配的感兴趣区域进行解析。在海马体中，我们发现 CA1a 中的浅层锥体神经元比深层的锥体神经元具有更多的棘突 aparati 密度。总的来说，这种方法可以使细胞和组织背景下的亚细胞特征的功能和结构研究成为可能。
