Spicer S S, Sens M A, Tashian R E
J Histochem Cytochem. 1982 Sep;30(9):864-73. doi: 10.1177/30.9.6813372.
Human tissues obtained early postmortem were immunostained to demonstrate carbonic anhydrase (CA) and, in some instances, to differentiate CA I and CA II, employing an immunoglobulin-peroxidase bridge method. Optimal immunostaining was obtained in tissues fixed a few hours in Carnoy's fluid or a buffered HgCl2 solution. Specimens fixed 1/2 to 2 hr with buffered formalin or Bouin's fluid stained less well but better than those fixed 24 hr with formalin. In tracheobronchial glands, serous acini and demilunes exhibited intense immunoreactivity demonstrative of the isozyme CA II. In kidney, all cells of the distal convoluted tubules were strongly positive for CA and cortical collecting tubule cells stained strongly but with some variability among individual cells. Cells in medullary collecting tubules ranged from intensely to negligibly reactive. Proximal convoluted tubules and thick ascending limbs showed moderate to light, uniform staining, but the thin limbs of the loop of Henle were negative. Renal cell immunoreactivity occurred only with antiserum to CA II. Seromucous acini in submandibular glands stained strongly and selectively for CA. Ducts in liver and pancreas showed strong selective immunostaining. The most superficial columnar cells lining the main lumen of the colon and appendix displayed strong reactivity, as did columnar cells lining the gall bladder.
对死后早期获取的人体组织进行免疫染色,以显示碳酸酐酶(CA),在某些情况下,采用免疫球蛋白-过氧化物酶桥法区分CA I和CA II。在卡诺氏液或缓冲氯化汞溶液中固定数小时的组织中获得了最佳免疫染色效果。用缓冲福尔马林或波因氏液固定1/2至2小时的标本染色效果较差,但比用福尔马林固定24小时的标本要好。在气管支气管腺中,浆液性腺泡和半月形细胞显示出强烈的免疫反应性,表明存在同工酶CA II。在肾脏中,远曲小管的所有细胞对CA呈强阳性,皮质集合小管细胞染色强烈,但个别细胞之间存在一些差异。髓质集合小管中的细胞反应性从强烈到可忽略不计不等。近曲小管和髓袢升支粗段显示中度至轻度的均匀染色,但髓袢细段呈阴性。肾细胞免疫反应仅与抗CA II血清发生。下颌下腺的浆液黏液性腺泡对CA染色强烈且具有选择性。肝脏和胰腺的导管显示出强烈的选择性免疫染色。结肠和阑尾主腔最表层的柱状细胞显示出强烈的反应性,胆囊内衬的柱状细胞也是如此。