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Öğe Blister artifact formation due to organic solvent effects on Spurr's epoxy resin semithin sections(Informa Healthcare, 1998) Kayton R.J.; Aktas R.G.Wrinkles and air bubble artifacts may occur when preparing slides of semithin sections (0.5 ?m) from blocks embedded in different resins. More than aesthetically annoying, wrinkles and air bubble artifacts may prohibit study of small structures. Present observations suggest that organic solvent based mounting media may interact with the resin of the section. This sometimes causes wrinkles and air bubble artifacts in the sections that degrade the quality of light microscope images. We compared the quality of semithin sections of several tissues in different resins using various types of mounting media. We observed that sections from Spurr's resin have many more artifacts. In particular, small, 2-10 ?m round or oblong blister-like artifacts often plague our Spurr's resin sections. We demonstrate that Spurr's resin sections react with toluene and xylene in organic solvent based mounting media forming blisters, while sections from Araldite and L. R. White do not. We suggest combinations of embedding and mounting media for successful preparation of semithin sections for light microscopy without wrinkles, blisters, or air bubble artifacts.Öğe Electron Microscopic Immunolocalization of Basic Fibroblast Growth Factor-Like Molecules in Capillary Endothelial Cells(Oxford University Press, 1998) Aktas R.G.; Kayton R.J.Basic fibroblast growth factor (bFGF) is a potent angiogenic polypeptide. It promotes angiogenesis in vivo and in vitro by stimulating migration, proliferation and proteolytic activity of endothelial cells. Whereas several effects of exogenous bFGF on endothelial cells have been described, it has remained unclear how endogenous bFGF produced by vascular endothelial cells regulate angiogenesis. To further investigate functional implications of the distribution of bFGF, we undertook the present study. Our aims were (i) to identify the specific location of bFGF in endothelial cells using electron microscopy immunogold labeling technique (ii) to determine the distribution of bFGF in capillaries of different types of tissues. Tissue samples from sciatic nerve, hippocampus, adrenal gland and kidney of normal adult rats were fixed in 4% paraformaldehyde/1 to 5% glutaraldehyde and embedded in Spurr's resin. Ultrathin sections were labeled with either polyclonal (F3393-Sigma) or monoclonal antibodies (F6162-Sigma, C3316-ZymoGenetics) specific for bFGF using a two-step immunogold labeling method. © 1998 Microscopy Society of America.Öğe Ultrastructural Distribution of Basic Fibroblast Growth Factor-Like Molecules in Peripheral Nerves(Oxford University Press, 1998) Kayton R.J.; Aktas R.G.Basic Fibroblast Growth Factor (bFGF) is a multifunctional polypeptide which has been shown to play a pivotal role in the survival and differentiation of nerve cells. Several trophic and non-trophic functions of this protein have been suggested in peripheral nerves. In spite of ample information about the distribution and effects of bFGF in central nervous system, few data are available concerning the localization of this protein in peripheral nerves. In view of the role of bFGF in regulation of trophic and non-trophic functions, we particularly focused on the presence and precise location of bFGF in peripheral nerves at the electron microscope level. Spurr's resin embedded ultrathin sections from adult rats' sural nerves were labeled with either polyclonal (F3393-Sigma) or monoclonal antibodies (F6162-Sigma, C3316-Zymogenetics) specific for bFGF using two-step immunogold labeling method. Control samples were treated with either an equivalent volume of blocking solution (omitting the primary antibody) or an irrelevant antibody (Factor VIII, VGF, anti-histamine, anti-fibroblast 5B5). © 1998 Microscopy Society of America.
