IMPROVEMENTS IN THE TECHNIQUE OF VASCULAR PERFUSION-FIXATION EMPLOYING A FLUOROCARBON-CONTAINING PERFUSATE AND A PERISTALTIC PUMP CONTROLLED BY PRESSURE FEEDBACK
IMPROVEMENTS IN THE TECHNIQUE OF VASCULAR PERFUSION-FIXATION EMPLOYING A FLUOROCARBON-CONTAINING PERFUSATE AND A PERISTALTIC PUMP CONTROLLED BY PRESSURE FEEDBACK
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DOI：
10.1111/j.1365-2818.1993.tb03405.x
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发表时间：
1993-11-01
影响因子：
2
通讯作者：
POULSEN, SS
中科院分区：
文献类型：
--
作者：
ROSTGAARD, J;QVORTRUP, K;POULSEN, SS
本文介绍了一种新的、改进的大鼠和其他小动物全身灌注固定技术。驱动力是一个蠕动泵，它由一个压力传感器进行反馈调节，该传感器监测心脏左心室中的血液/灌注压力。主要灌注液固定剂由血液替代品-含2%戊二醛的0.05 M二甲胂酸盐缓冲液（pH 7.4）中的13.3%含氧氟碳FC-75组成。二级灌注液固定剂由2%戊二醛的0.05 M二甲胂酸盐缓冲液（pH 7.4）和20 mM CaCl 2组成。双筒、自保持插管用于心脏插管;插管的外筒和内筒分别连接到蠕动泵和压力传感器。通过皮下氧电极监测大鼠的组织氧张力。测量结果表明，组织缺氧/缺氧没有发展之前或期间的灌注固定。因此，该技术允许研究不显示固定梯度并且不包含在窒息状态下固定的细胞的标本。不同器官细胞的电子显微照片证实了这一点，揭示了新的精细结构元素。通过将含氧氟碳化合物添加到戊二醛灌注固定剂中，为细胞呼吸以及戊二醛与组织的耗氧化学反应提供足够的氧气。麻醉，手术操作，机械组件的系统，准备的固定剂和流动的灌注液固定剂的数据提供和讨论。
A new, improved technique for whole-body perfusion-fixation of rats and other small animals is described. The driving force is a peristaltic pump which is feedback regulated by a pressure transducer that monitors the blood/perfusion pressure in the left ventricle of the heart. The primary perfusate-fixative is composed of a blood substitute-13.3% oxygenated fluorocarbon FC-75-in 0.05 m cacodylate buffer (pH 7.4) with 2% glutaraldehyde. The secondary perfusate-fixative is composed of 2% glutaraldehyde in 0.05 M cacodylate buffer (pH 7.4) with 20 mM CaCl2. A double-barrelled, self-holding cannula is used to cannulate the heart; the outer and inner barrels of the cannula are connected to the peristaltic pump and to the pressure transducer, respectively. The tissue oxygen tension in the rat is monitored by a subcutaneous oxygen electrode. Measurements showed that tissue hypoxia/anoxia did not develop before or during the perfusion-fixation. Thus, the technique permits study of specimens which do not exhibit fixation gradients and do not contain cells fixed in a state of asphyxia. This is substantiated by electron micrographs of cells from different organs, revealing new fine structural elements. By adding oxygenated fluorocarbon to glutaraldehyde perfusate-fixatives, enough oxygen is made accessible for cellular respiration as well as for the oxygen-consuming chemical reactions of glutaraldehyde with the tissue. Data on anaesthesia, operative manoeuvres, mechanical components of the system, preparation of fixatives and flow of the perfusate-fixatives are furnished and discussed.