Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Micron. 2011 Dec;42(8):762-72. doi: 10.1016/j.micron.2011.06.005. Epub 2011 Jun 25.
Biological specimens have to be prepared for imaging in the electron microscope in a way that preserves their native structure. Two-dimensional (2D) protein crystals to be analyzed by electron crystallography are best preserved by sugar embedding. One of the sugars often used to embed 2D crystals is trehalose, a disaccharide used by many organisms for protection against stress conditions. Sugars such as trehalose can also be added to negative staining solutions used to prepare proteins and macromolecular complexes for structural studies by single-particle electron microscopy (EM). In this review, we describe trehalose and its characteristics that make it so well suited for preparation of EM specimens and we review specimen preparation methods with a focus on the use of trehalose.
生物标本必须经过特殊处理,以在电子显微镜下保持其天然结构。二维(2D)蛋白晶体通过电子晶体学进行分析,最好用糖包埋来保存。在用于电子显微镜(EM)单颗粒技术研究的负染色溶液中,经常使用海藻糖等糖来包埋 2D 晶体,海藻糖是许多生物用来抵御压力条件的一种二糖。在本文中,我们描述了海藻糖及其特性,使其非常适合用于制备 EM 标本,并重点介绍了使用海藻糖的标本制备方法。