SINGLE TUBULE PERFUSION TECHNIQUES
SINGLE TUBULE PERFUSION TECHNIQUES
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DOI：
10.1038/ki.1986.168
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发表时间：
1986-08-01
影响因子：
19.6
通讯作者：
KNEPPER, MA
中科院分区：
文献类型：
--
作者：
BURG, MB;KNEPPER, MA
20年前，一种分离肾单位节段并在体外灌流的技术被引入[1]。从那时起，基于这项技术的结果，已经发表了数百篇论文。对该程序的技术细节[2，31，其历史[41，以及结果本身[5，6]进行了广泛的审查。在这篇文章中，我们将局限于这项技术的一些一般方面，即可以提供的信息类型、优点和局限性，以及获得健康图尔克的方法。实验的设计，结果的解释，以及新的发展和趋势。为了在体外灌流肾小管，从肾脏分离出肾小管，一端连接到玻璃微管以灌流管腔，自由端连接到另一微管以收集灌流的液体。以这种方式，管腔有效地与浴缸隔离。在灌流过程中以受控的速度收集液体，并进行分析，以确定与灌流液相比成分的差异。根据这些结果，可以计算出输运率和极限浓度梯度。电生理学技术可以用来测量跨上皮和细胞的电压和电阻。细胞的组成可以在离子敏感微电极的灌流过程中测量[8]，也可以通过灌流后组织的化学分析来测量。细胞形态也可以在灌流过程中用光学显微镜检查，或在灌流后用电子显微镜固定检查。
Twenty years ago a technique for isolating nephron segments and perfusing them in vitro was introduced [1]. Since then hundreds of papers have been published based on results from the technique. There have been extensive reviews of the technical details of the procedure [2, 31, its history [41, and of the results themselves [5, 6]. In this paper we will restrict ourselves to some general aspects of the technique, namely the types of information that can he provided, advantages and limitations, methods for obtaining healthy tuhulcs. design of experiments, interpretation of results, and new developments and trends.To perfuse renal tubules in vitro, they are dissected from the kidney, one end is connected to a glass micropipet to perfuse the lumen, and the free end is connected to another micropipet to collect the perfused fluid. in this fashion the tubule lumen is effectively isolated from the bath. Fluid is collected during perfusion at a controlled rate and analyzed to determine differences in composition compared to the perfusate. From these results, transport rates and limiting concentration gradients can be calculated. Electrophysiological techniques can he used to measure transepithelial and cellular voltages and resistances [71. The composition of the cells can be measured either during perfusion with ion sensitive microelectrodes [8] or by chemical analysis of the tissue after perfusion. Cellular morphology also can he examined either during perfusion by light microscopy or after perfusion by fixation for electron microscopy.