Ultrastructural Analysis of Testicular Tissue and Sperm by Transmission and Scanning Electron Microscopy
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.
Key wordsTransmission electron microscopy Ultrastructural immunocytochemistry Ultrastructural histochemistry Scanning electron microscopy Testes Semen Spermatozoa Spermatogenesis
Supported by Grants from CONICET (PIP 112200901 00615) and ANPCyT (PICT 2005 38229). Figures 6 and 1 are modified from refs. 12 and 17, Fig. 4d is taken from ref. 15, and Fig. 2d is taken from ref. 18 with permission (see figure legends).
The author is indebted to Prof. I. von Lawzewitsch and R. Mancini from the Histology Department, Buenos Aires University and Prof. Don W. Fawcett and all staff from the Anatomy Department, Harvard Medical School where his initial training in electron microscopy took place. Special thanks to R. Burghardt Ph.D., M. Musse M.Sc., and other colleagues with whom the author has collaborated over the years.
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