Urban K, Hewicker-Trautwein M
Institut für Pathologie, Tierärztlichen Hochschule Hannover, Germany.
Acta Histochem. 1994 Dec;96(4):365-77. doi: 10.1016/S0065-1281(11)80020-6.
The immunohistochemical staining of vimentin in paraffin-embedded sections from adult cattle, rabbit, rat and mouse brain fixed in different fixatives (formaldehyde, methacarn, ethanol) was examined using two monoclonal antibodies and a polyclonal antiserum. In non-trypsinized formaldehyde-fixed tissue sections both monoclonal antibodies and the polyclonal antibodies failed to stain vimentin. Following trypsinization of formaldehyde-fixed sections of the four species the meninges, endothelial cells of blood vessels, ependymal cells and the stroma of the choroid plexus were labelled by the monoclonal and polyclonal antibodies used. Astrocytes and Bergmann glial fibers in pretrypsinized formaldehyde-fixed sections from cattle, rabbit and rat brain, however, showed only weak staining. Fixation of cattle and rat brain in methacarn markedly improved the vimentin immunoreactivity of astrocytes and Bergmann glial fibers. The best fixative for the preservation of immunoreactive determinants of vimentin in astrocytes and Bergmann glial fibers in cattle, rabbit and rat brain was ethanol. In brain tissue from mice both monoclonal antibodies labelled only mesoderm-derived tissue components, but did not recognize vimentin in astrocytes and Bergmann glial fibers. Pre-heating formaldehyde-fixed sections from cattle, rabbit and rat brain in a microwave oven prior to the immunohistochemical reaction resulted in an enormous enhancement of vimentin staining of mesoderm-derived tissues, of astrocytes and bergmann cell fibers.
使用两种单克隆抗体和一种多克隆抗血清，检测了用不同固定剂（甲醛、甲醇 Carnoy 固定液、乙醇）固定的成年牛、兔、大鼠和小鼠脑石蜡包埋切片中波形蛋白的免疫组织化学染色。在未经胰蛋白酶处理的甲醛固定组织切片中，单克隆抗体和多克隆抗体均未能对波形蛋白进行染色。对这四种动物的甲醛固定切片进行胰蛋白酶处理后，所用的单克隆抗体和多克隆抗体标记了脑膜、血管内皮细胞、室管膜细胞和脉络丛基质。然而，在牛、兔和大鼠脑甲醛固定前未经胰蛋白酶处理的切片中，星形胶质细胞和贝格曼胶质纤维仅显示弱阳性染色。用甲醇 Carnoy 固定液固定牛和大鼠脑，显著改善了星形胶质细胞和贝格曼胶质纤维的波形蛋白免疫反应性。保存牛、兔和大鼠脑星形胶质细胞和贝格曼胶质纤维中波形蛋白免疫反应性决定簇的最佳固定剂是乙醇。在小鼠脑组织中，两种单克隆抗体仅标记中胚层来源的组织成分，而未识别星形胶质细胞和贝格曼胶质纤维中的波形蛋白。在免疫组织化学反应之前，将牛、兔和大鼠脑甲醛固定切片在微波炉中预热，导致中胚层来源组织、星形胶质细胞和贝格曼细胞纤维的波形蛋白染色显著增强。
