Electron Microscopy Facility-Instituto Gulbenkian de Ciência, Oeiras, Portugal.
Histopathology Facility-Instituto Gulbenkian de Ciência, Oeiras, Portugal.
Methods Cell Biol. 2021;162:13-37. doi: 10.1016/bs.mcb.2020.12.001. Epub 2021 Feb 18.
Bridging from the macrostructure to the nanostructure of tissues is often technically challenging. To try to solve this, we developed a flexible CLEM workflow that can be applied to the analysis of tissues from diverse model organisms across various length scales. The Histo-CLEM Workflow combines three main microscopy techniques, namely histology, light microscopy and electron microscopy. Herein, all the steps of the Histo-CLEM Workflow are explained in detail to enable the adaptation of the method to tissue particularities and biological questions. The preparation and visualization of mice nerve fibers is shown as an application example of the presented Histo-CLEM Workflow.
从宏观结构到组织的纳米结构的桥接通常在技术上具有挑战性。为了解决这个问题,我们开发了一种灵活的 CLEM 工作流程,可以应用于不同模型生物和各种长度尺度的组织分析。Histo-CLEM 工作流程结合了三种主要的显微镜技术,即组织学、光学显微镜和电子显微镜。本文详细解释了 Histo-CLEM 工作流程的所有步骤,以实现该方法对组织特殊性和生物学问题的适应。本文以展示的 Histo-CLEM 工作流程为例,介绍了小鼠神经纤维的制备和可视化。