Heiligenstein Xavier, Paul-Gilloteaux Perrine, Raposo Graça, Salamero Jean
Institut Curie, PSL Research University, CNRS UMR 144 & Cell and Tissue Imaging Facility, Paris, France.
Structure Fédérative de Recherche François Bonamy, INSERM, CNRS, Université de Nantes, Nantes, France.
Methods Cell Biol. 2017;140:335-352. doi: 10.1016/bs.mcb.2017.03.014. Epub 2017 Apr 19.
Correlative light and electron microscopy (CLEM) is a scientific method covered by a broad range of techniques. The path taken to explore a scientific question is often driven both by the question and the technology available. Yet, one common step to all CLEM workflows is the registration of the multimodal images to assign a fluorescent signal to an ultrastructure. The manual relocation and registration of light microscopy and electron microscopy images can be challenging and time-consuming (Muller-Reichert & Verkade, 2014). eC-CLEM is a free open-source software to address this step. eC-CLEM has been designed with an intuitive procedure and the manual registration has been extensively described in step-by-step protocols on the eC-CLEM webpage as well as video tutorials. In this book chapter, we focus our description on the "automatic registration" procedure, which requires some fine tuning. We recommend the user to first get familiar with eC-CLEM through the aforementioned tutorials. If large volume data sets or automatic tracking and controlling of microscopes are pursued by the user, going through the fine-tuning steps described in this chapter is worth the effort.
相关光电子显微镜（CLEM）是一种涵盖多种技术的科学方法。探索科学问题所采用的途径通常由问题本身和可用技术共同驱动。然而，所有CLEM工作流程的一个共同步骤是对多模态图像进行配准，以便将荧光信号与超微结构对应起来。光学显微镜和电子显微镜图像的手动重新定位和配准可能具有挑战性且耗时（Muller-Reichert & Verkade，2014）。eC-CLEM是一款用于解决此步骤的免费开源软件。eC-CLEM的设计流程直观，其手动配准在eC-CLEM网页上的分步协议以及视频教程中都有详细描述。在本章中，我们将重点描述“自动配准”过程，该过程需要一些微调。我们建议用户首先通过上述教程熟悉eC-CLEM。如果用户需要处理大量数据集或对显微镜进行自动跟踪和控制，那么仔细研读本章所述的微调步骤是值得的。
