裂殖酵母的电子显微镜观察

Electron Microscopy of Fission Yeast.

作者信息

McIntosh J Richard, Morphew Mary K, Giddings Thomas H

机构信息

Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347.

Laboratory for 3D Electron Microscopy, University of Colorado, Boulder, Colorado 80309-0347.

出版信息

Cold Spring Harb Protoc. 2017 Jan 3;2017(1):2017/1/pdb.top079822. doi: 10.1101/pdb.top079822.

DOI:10.1101/pdb.top079822

PMID:28049809

Abstract

Electron microscopy (EM) can provide images of cells with a spatial resolution that significantly surpasses that available from light microscopy (LM), even with modern methods that give LM "super resolution." However, EM resolution comes with costs in time spent with sample preparation, expense of instrumentation, and concerns regarding sample preparation artifacts. It is therefore important to know the limitations of EM as well as its strengths. Here we describe the most reliable methods for the preservation of fission yeast cells currently available. We describe the properties of images obtained by transmission EM (TEM) and contrast them with images from scanning EM (SEM). We also show how one can make three-dimensional TEM images and discuss several approaches to address the problem of localizing specific proteins within cells. We give references to work by others who have pursued similar goals with different methods, and we discuss briefly the complex subject of image interpretation.

摘要

电子显微镜（EM）能够提供细胞图像，其空间分辨率显著超过光学显微镜（LM），即便采用使LM具备“超分辨率”的现代方法亦是如此。然而，EM分辨率在样本制备耗时、仪器成本以及对样本制备假象的担忧方面存在代价。因此，了解EM的局限性及其优势很重要。在此，我们描述目前可用的保存裂殖酵母细胞的最可靠方法。我们描述通过透射电子显微镜（TEM）获得的图像特性，并将其与扫描电子显微镜（SEM）的图像进行对比。我们还展示了如何制作三维TEM图像，并讨论了几种解决在细胞内定位特定蛋白质问题的方法。我们引用了其他采用不同方法追求类似目标的研究者的工作，并简要讨论图像解释这一复杂主题。