Hollenberg M J, Lea P J
Department of Anatomy, Faculty of Medicine, University of Toronto, Ontario, Canada.
Invest Ophthalmol Vis Sci. 1988 Sep;29(9):1380-9.
The application of a new technique in vision research for investigating the three-dimensional ultrastructure of extra- and intracellular organelles and components is presented, based on the use of high resolution scanning electron microscopy (HRSEM). This technique has been made possible by advances in scanning electron microscope design which permit resolution of particles less than 3 nm in diameter and new tissue preparation techniques which remove the cytosol and leave cell membranes and organelles in relief. Application of these methods to the ultrastructure of the retinal pigment epithelium and Bruch's layer in the albino rat has revealed a number of new structural features and confirmed others by using a different approach in tissue preparation and examination. It is now possible to obtain HRSEM micrographs with an information content equivalent to at least 15 ultrathin, serial, transmission electron microscope sections, all perfectly aligned, in one micrograph without having to resort to accepted reconstruction techniques. It is clear that this new method has the potential to rapidly advance our knowledge of the structures involved in vision in both health and disease and in a variety of experimental conditions.
本文介绍了一种基于高分辨率扫描电子显微镜（HRSEM）的新技术在视觉研究中的应用，该技术用于研究细胞外和细胞内细胞器及组分的三维超微结构。扫描电子显微镜设计的进步使得该技术成为可能，这些进步允许分辨直径小于3nm的颗粒，同时新的组织制备技术能够去除细胞质溶胶，使细胞膜和细胞器凸显出来。将这些方法应用于白化大鼠视网膜色素上皮和布鲁赫膜的超微结构研究，通过采用不同的组织制备和检查方法，揭示了许多新的结构特征并证实了其他一些特征。现在可以获得信息含量相当于至少15个超薄连续透射电子显微镜切片的HRSEM显微照片，所有切片都完美对齐在一张显微照片中，而无需借助公认的重建技术。显然，这种新方法有潜力在健康和疾病状态以及各种实验条件下，迅速推进我们对视觉相关结构的认识。
