Karreman Matthia A, Ruthensteiner Bernhard, Mercier Luc, Schieber Nicole L, Solecki Gergely, Winkler Frank, Goetz Jacky G, Schwab Yannick
European Molecular Biology Laboratory, Heidelberg, Germany.
Zoologische Staatssammlung München, Munich, Germany.
Methods Cell Biol. 2017;140:277-301. doi: 10.1016/bs.mcb.2017.03.006. Epub 2017 Apr 21.
Combining in vivo imaging with electron microscopy (EM) uniquely allows monitoring rare and critical events in living tissue, followed by their high-resolution visualization in their native context. A major hurdle, however, is to keep track of the region of interest (ROI) when moving from intravital microscopy (IVM) to EM. Here, we present a workflow that relies on correlating IVM and microscopic X-ray computed tomography to predict the position of the ROI inside the EM-processed sample. The ROI can then be accurately and quickly targeted using ultramicrotomy and imaged using EM. We outline how this procedure is used to retrieve and image tumor cells arrested in the vasculature of the mouse brain.
将体内成像与电子显微镜（EM）相结合，能够独特地监测活组织中罕见且关键的事件，随后在其原生环境中对这些事件进行高分辨率可视化。然而，一个主要障碍是，当从活体显微镜检查（IVM）过渡到EM时，要跟踪感兴趣区域（ROI）。在此，我们提出一种工作流程，该流程依赖于将IVM与显微X射线计算机断层扫描相关联，以预测EM处理样本内ROI的位置。然后，可以使用超薄切片技术准确、快速地定位ROI，并使用EM进行成像。我们概述了如何使用此程序来检索并成像停滞在小鼠脑血管中的肿瘤细胞。
