Graham Lesley, Orenstein Jan Marc
Department of Pathology, George Washington University Medical Center, Ross 502, 2300 Eye Street, NW, Washington, District of Columbia 20037, USA.
Nat Protoc. 2007;2(10):2439-50. doi: 10.1038/nprot.2007.304.
In transmission electron microscopy (TEM), electrons are transmitted through a plastic-embedded specimen, and an image is formed. TEM enables the resolution and visualization of detail not apparent via light microscopy, even when combined with immunohistochemical analysis. Ultrastructural examination of tissues, cells and microorganisms plays a vital role in diagnostic pathology and biologic research. TEM is used to study the morphology of cells and their organelles, and in the identification and characterization of viruses, bacteria, protozoa and fungi. In this protocol, we present a TEM method for preparing specimens obtained in clinical or research settings, discussing the particular requirements for tissue and cell preparation and analysis, the need for rapid fixation and the possibility of analysis of tissue already fixed in formalin or processed into paraffin blocks. Details of fixation, embedding and how to prepare thin and semi-thin sections, which can be used for analysis complementary to that performed ultimately using TEM, are also described.
在透射电子显微镜（TEM）中，电子透过塑料包埋的标本，从而形成图像。即使与免疫组织化学分析相结合，TEM也能够分辨并呈现出光学显微镜下不明显的细节。对组织、细胞和微生物进行超微结构检查在诊断病理学和生物学研究中起着至关重要的作用。TEM用于研究细胞及其细胞器的形态，以及鉴定和表征病毒、细菌、原生动物和真菌。在本方案中，我们介绍一种用于制备临床或研究环境中所获标本的TEM方法，讨论组织和细胞制备及分析的特殊要求、快速固定的必要性以及对已用福尔马林固定或制成石蜡块的组织进行分析的可能性。文中还描述了固定、包埋的细节，以及如何制备可用于与最终使用TEM进行的分析互补的薄切片和半薄切片。
