PERFUSION FIXATION OF LUNGS FOR STRUCTURE-FUNCTION ANALYSIS - CREDITS AND LIMITATIONS
PERFUSION FIXATION OF LUNGS FOR STRUCTURE-FUNCTION ANALYSIS - CREDITS AND LIMITATIONS
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DOI：
10.1152/jappl.1982.53.2.528
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发表时间：
1982-01-01
影响因子：
3.3
通讯作者：
WEIBEL, ER
中科院分区：
文献类型：
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作者：
BACHOFEN, H;AMMANN, A;WEIBEL, ER
对血管灌注固定的肺组织保存质量进行了重新评价。用不同但广泛使用的固定剂灌注膨胀至总肺容量60%的离体兔肺（III区条件）。通过连续监测评估灌注液对相关生理变量的影响;通过EM的定性和形态测定分析评估对肺微观结构的影响。在生理范围内的流速下用等渗戊二醛进行灌注，会导致灌注压和肺重量大幅增加，这反映了细胞内、间质和肺泡内水肿。如果将戊二醛加入等渗缓冲液（总渗透压510 mosM [毫渗摩尔]）中，则不会发生水肿。戊二醛作为唯一灌注液不能完全消除肺组织的收缩力。在释放经肺压力时，肺以无法确定的量缩回。用OsO_4和醋酸铀酰或戊二醛（510 mosM），然后用OsO_4和醋酸铀酰顺序灌注，可以获得令人满意的结果。后一种组合产生研究肺泡和毛细血管结构的最佳制剂，但引起肺细胞的高渗体积损失（细胞收缩）。
The quality of tissue preservation in lungs fixed by vascular perfusion was reevaluated. Excised rabbit lungs inflated to 60% of total lung capacity were perfused (zone III conditions) with different but widely used fixatives. Effects of the perfusates on pertinent physiological variables were assessed by continuous monitoring; the effects on the pulmonary microstructure, by qualitative and morphometric analysis of EM. Perfusions with isotonic glutaraldehyde at flow rates within the physiological range produce large increases of perfusion pressure and lung weight that reflect intracellular, interstitial and intraalveolar edema. No edema occurs if glutaraldehyde is added to isotonic buffer solutions (total osmolarity 510 mosM [milliosmole]). Glutaraldehyde as sole perfusate does not fully eliminate the retractive force of lung tissue. Upon release of transpulmonary pressure, the lungs retract by an indeterminable amount. Satisfactory results can be obtained by sequential perfusion with OsO4 and uranyl acetate or glutaraldehyde (510 mosM) followed by OsO4 and uranyl acetate. The latter combination yields optimal preparations to study the alveolar and capillary architecture but causes a hyperosmotic volume loss of lung cells (cell shrinkage).