Schaub R G, Rawlings C A, Stewart G J
Am J Vet Res. 1980 Sep;41(9):1441-6.
In this communication we describe (1) a method for preparing pulmonary arteries and veins for examination by scanning electron microscopy and (2) the topographic characteristics of the luminal surface of these vessels. Seven healthy adult dogs were used in this study. The pulmonary vasculature was washed free of blood by perfusion with Tyrode's solution. Blood vessels were partially fixed in situ under physiologic pressure in 3 dogs (15 mm of Hg) or 2 times physiologic pressure in 4 dogs (30 mm of Hg) by perfusion with 1% glutaraldehyde. The pulmonary artery and aorta were tied off and the lungs were removed and immersed in 1% glutaraldehyde for 4 hours for additional fixation under pressure. Blood vessels were dissected free of surrounding tissue, further fixed in 2.5% glutaraldehyde, postfixed in osmium tetroxide, dehydrated, and critical point dried. The luminal surfaces were sufficiently flat to allow visualization of most of the topographic features of the luminal surface. Even 1 missing endothelial cell wound have been obvious. Contrary to previous reports, we did not find a dense population of finger-like projections on the surface of arterial endothelium. Rather, we found a modest number of small rounded blebs. The venous endothelial sheet was similar in appearance. These observations demonstrate that scanning electron microscopy may be used to study vascular lesions in pulmonary vaessels, as well as in more accessible peripheral vessels.
在本报告中,我们描述了(1)一种制备肺动脉和肺静脉以用于扫描电子显微镜检查的方法,以及(2)这些血管腔表面的地形特征。本研究使用了7只健康成年犬。通过用台氏液灌注将肺血管中的血液冲洗干净。在3只犬中通过灌注1%戊二醛在生理压力(15 mmHg)下对血管进行原位部分固定,在4只犬中在2倍生理压力(30 mmHg)下进行固定。结扎肺动脉和主动脉,取出肺并浸入1%戊二醛中4小时以在压力下进行额外固定。将血管从周围组织中分离出来,进一步固定于2.5%戊二醛中,用四氧化锇后固定,脱水,然后临界点干燥。管腔表面足够平坦,能够观察到管腔表面的大部分地形特征。甚至单个缺失的内皮细胞伤口也很明显。与先前的报道相反,我们在动脉内皮表面未发现密集的指状突起。相反,我们发现了少量小的圆形泡。静脉内皮片外观相似。这些观察结果表明,扫描电子显微镜可用于研究肺血管以及更容易观察的外周血管中的血管病变。