Light and electron microscopy of virus-associated, intranuclear paracrystals in cultured cells infected with types 2,4,6, and 18 human adenoviruses.
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A new technique for the cytochemical demonstration of peroxidase is presented. Monomeric homovanillic acid is converted by H2O2 and peroxidase into its dimeric form, which is then precipitated as a complex salt of lead and rhodamine 6G or rhodamine B. The reaction product can be visualized by conversion to lead sulphide or viewed directly under the fluorescence microscope, since it emits a red fluorescence when excited with green light. The reaction is rapid and results in good localization at the cytologic level of peroxidase activity in granulocytes. The technique can be applied for the ultrastructural localization of enzymatic activity, but in its present form it does not match the localization sharpness of the diaminobenzidine method. This fluorescent cytochemical technique will also detect horseradish peroxidase activity and may provide a usefull probe in peroxidase immunohistochemistry. The principle of complexing metal salts with fluorescent dyes may find a more general application in enzyme cytochemistry.
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A method has been developed to enable correlative light microscopy (LM) and scanning electron microscopy (SEM) on the same section of wheat (Triticum aestivum L.) leaves infested by greenbug aphids (Schizaphis graminum Rondani). Segments of infested leaf tissue were fixed, embedded in paraffin, sectioned, and affixed to slides by standard histological techniques. Serial sections were viewed by LM as temporary mounts in xylene. Sections of interest were identified and re-embedded in fingernail polish, affixed to aluminum stubs, freed of polish with ethyl acetate or acetone, and sputter-coated for SEM. SEM of re-embedded leaf sections showed excellent preservation of leaf anatomy. The same aphid tracks and regions of cell damage identified by LM were visible. SEM increased resolution and provided a much clearer sense of the three-dimensional relations involved in the interaction between plant and insect.
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This report describes the intracellular and extracellular localization of fibronectin at the ultrastructural level in primary cultures of aortic smooth muscle cells. Fibronectin was present in all the cisternae of the rough endoplasmic reticulum except the perinuclear cisterna, in large vesicles associated with the trans side of the Golgi complex, and in single large vesicles in the cytoplasm often associated with microtubules. The extracellular microfibrils were heavily stained. In sections parallel to the plane of growth bundles of extracellular microfibrils in continuity with arrays of intracellular microfilaments were observed (fibronexus). The basement membrane around the aortic smooth muscle cells was discontinuous and diffusely stained. The results indicate that fibronectin is localized in the cytoplasmic membranous apparatus of protein synthesis, processing, and secretion. The lack of reaction product in the flat cisternae of the Golgi complex let suggest either that fibronection may not be present in significant amounts within the flattened cisternae or that the method is insufficient in detecting the glycoprotein in this subcompartment off the Golgi complex.