Department of Pathology and Cancer Research and Treatment Center, University of New Mexico, Albuquerque, NM 87131, USA.
J Cell Sci. 2012 Jun 1;125(Pt 11):2571-80. doi: 10.1242/jcs.090027. Epub 2012 Jun 8.
A fundamental goal in biology is to determine how cellular organization is coupled to function. To achieve this goal, a better understanding of organelle composition and structure is needed. Although visualization of cellular organelles using fluorescence or electron microscopy (EM) has become a common tool for the cell biologist, recent advances are providing a clearer picture of the cell than ever before. In particular, advanced light-microscopy techniques are achieving resolutions below the diffraction limit and EM tomography provides high-resolution three-dimensional (3D) images of cellular structures. The ability to perform both fluorescence and electron microscopy on the same sample (correlative light and electron microscopy, CLEM) makes it possible to identify where a fluorescently labeled protein is located with respect to organelle structures visualized by EM. Here, we review the current state of the art in 3D biological imaging techniques with a focus on recent advances in electron microscopy and fluorescence super-resolution techniques.
生物学的一个基本目标是确定细胞组织如何与功能相偶联。为了实现这一目标，需要更好地了解细胞器的组成和结构。虽然使用荧光或电子显微镜（EM）可视化细胞细胞器已经成为细胞生物学家的常用工具，但最近的进展比以往任何时候都更清楚地描绘了细胞。特别是，先进的光学显微镜技术正在实现低于衍射极限的分辨率，而 EM 断层扫描提供了细胞结构的高分辨率三维（3D）图像。能够在同一样品上同时进行荧光和电子显微镜（相关光和电子显微镜，CLEM）的能力使得可以确定荧光标记蛋白相对于通过 EM 可视化的细胞器结构的位置。在这里，我们回顾了 3D 生物成像技术的最新进展，重点介绍了电子显微镜和荧光超分辨率技术的最新进展。
