Pierson Jason, Vos Matthijn, McIntosh J Richard, Peters Peter J
Division of Cell Biology II, The Netherlands Cancer Institute - Antoni van Leeuwenhoek Hospital, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
J Electron Microsc (Tokyo). 2011;60 Suppl 1(Suppl 1):S93-100. doi: 10.1093/jmicro/dfr014.
A major objective of modern structural biology is to appreciate the cellular organization by elucidating the spatial arrangement of macromolecular complexes within a cell. Cryogenic sample preparation, combined with cryo-ultramicrotomy, enables large cells and pieces of biological tissues to be thinned for electron cryo-tomography, which provides a three-dimensional view of the biological sample. There are, however, limitations associated with the technique that must be realized, addressed and overcome for the procedure to become mainstream. Here, we provide perspectives on the continued advancements in cryogenic sample preparation for vitreous cryo-sectioning, image collection and post-image processing that have expanded the attainable information limit within the three-dimensional reconstructions of cells and pieces of biological tissues.
现代结构生物学的一个主要目标是通过阐明细胞内大分子复合物的空间排列来了解细胞组织。低温样品制备与低温超薄切片术相结合，能够将大细胞和生物组织切片变薄，用于电子冷冻断层扫描，从而提供生物样品的三维视图。然而，该技术存在一些局限性，必须认识到、解决并克服这些局限性，该方法才能成为主流。在此，我们对玻璃化冷冻切片的低温样品制备、图像采集和图像后处理方面的持续进展提供见解，这些进展扩大了细胞和生物组织切片三维重建中可获得的信息极限。
