Aliev G M, Mironov A A
Arkh Anat Gistol Embriol. 1990 Dec;99(12):31-6.
By means of scanning and transmissive electron microscopy structural peculiarities of endothelium of the thoracic and abdominal parts of the intact rat aorta have been studied at various regimens of preparation and making specimens . The greatest changes endotheliocytes (EC) undergo at using immersion fixation after dissection of the aortal segments. These changes are less pronounced at immersion fixation in situ. A decreased perfusion pressure results in appearance of intimal folds and microfolds on the surface of EC. Increasing time for washing more than 1 min results in appearance of inflations and craters on the surfice of EC. For analysis by means of transmissive electron microscopy it is not necessary to remove blood completely out of the vascular bed. The most essential factor is to maintain perfusion pressure at the average systolic level in the given area of the vessel. However, to make the analysis by means of scanning electron microscopy this method is not suitable. The most optimal condition for initial stages of preparing vessels for morphological investigation is their washing for 1 min in the medium 199 with addition of heparin (10 units/ml) during no more than 1 min with a subsequent perfusive fixation in 2.5% solution of glutaraldehyde in the medium 199 no less than 5 min under the average arterial pressure in the given area of the vessel.
通过扫描电子显微镜和透射电子显微镜,研究了完整大鼠主动脉胸段和腹段内皮在不同标本制备方案下的结构特点。在主动脉段解剖后采用浸入固定时,内皮细胞(EC)发生的变化最大。原位浸入固定时这些变化则不太明显。灌注压力降低会导致EC表面出现内膜褶皱和微褶皱。冲洗时间超过1分钟会导致EC表面出现膨隆和凹陷。用透射电子显微镜分析时,不必将血管床中的血液完全排空。最关键的因素是在血管的给定区域将灌注压力维持在平均收缩压水平。然而,用扫描电子显微镜进行分析时,这种方法并不适用。对血管进行形态学研究的初始阶段,最理想的条件是在含肝素(10单位/毫升)的199培养基中冲洗1分钟,冲洗时间不超过1分钟,随后在血管给定区域的平均动脉压下,用2.5%戊二醛的199培养基进行灌注固定不少于5分钟。