Chemes Hector E
Center for Research in Endocrinology (CEDIE), National Research Council (CONICET), Buenos Aires Children's Hospital, Buenos Aires, Argentina.
Methods Mol Biol. 2013;927:321-48. doi: 10.1007/978-1-62703-038-0_29.
Transmission electron microscopy (TEM) studies have provided the basis for an in-depth understanding of the cell biology and normal functioning of the testis and male gametes and have opened the way to characterize the functional role played by specific organelles in spermatogenesis and sperm function. The development of the scanning electron microscope (SEM) extended these boundaries to the recognition of cell and organ surface features and the architectural array of cells and tissues. The merging of immunocytochemical and histochemical approaches with electron microscopy has completed a series of technical improvements that integrate structural and functional features to provide a broad understanding of cell biology in health and disease. With these advances the detailed study of the intricate structural and molecular organization as well as the chemical composition of cellular organelles is now possible. Immunocytochemistry is used to identify proteins or other components and localize them in specific cells or organelles with high specificity and sensitivity, and histochemistry can be used to understand their function (i.e., enzyme activity). When these techniques are used in conjunction with electron microscopy their resolving power is further increased to subcellular levels. In the present chapter we will describe in detail various ultrastructural techniques that are now available for basic or translational research in reproductive biology and reproductive medicine. These include TEM, ultrastructural immunocytochemistry, ultrastructural histochemistry, and SEM.
透射电子显微镜（TEM）研究为深入理解睾丸和雄性配子的细胞生物学及正常功能提供了基础，并为表征特定细胞器在精子发生和精子功能中所起的功能作用开辟了道路。扫描电子显微镜（SEM）的发展将这些边界扩展到对细胞和器官表面特征以及细胞和组织的结构排列的识别。免疫细胞化学和组织化学方法与电子显微镜的结合完成了一系列技术改进，这些改进整合了结构和功能特征，以全面了解健康和疾病状态下的细胞生物学。随着这些进展，现在可以对细胞器复杂的结构和分子组织以及化学成分进行详细研究。免疫细胞化学用于识别蛋白质或其他成分，并以高特异性和敏感性将它们定位在特定细胞或细胞器中，组织化学可用于了解它们的功能（即酶活性）。当这些技术与电子显微镜结合使用时，它们的分辨能力进一步提高到亚细胞水平。在本章中，我们将详细描述目前可用于生殖生物学和生殖医学基础研究或转化研究的各种超微结构技术。这些技术包括透射电子显微镜、超微结构免疫细胞化学、超微结构组织化学和扫描电子显微镜。
