Quantifying Single Microvessel Permeability in Isolated Blood-perfused Rat Lung Preparation
Quantifying Single Microvessel Permeability in Isolated Blood-perfused Rat Lung Preparation
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DOI：
10.3791/51552
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发表时间：
2014-06-01
影响因子：
1.2
通讯作者：
Parthasarathi, Kaushik
中科院分区：
文献类型：
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作者：
Kandasamy, Kathirvel;Parthasarathi, Kaushik
分离的血液灌注肺制备被广泛用于可视化和定义单个微血管中的信号传导。通过将该制备与真实的时间成像相结合，确定个体肺微血管中的渗透性变化变得可行。在此，我们描述了分离大鼠肺和用自体血灌注它们的步骤。然后，我们概述了通过微导管将荧光团或试剂注入小的肺部区域的步骤。使用所描述的这些程序，我们确定了大鼠肺微血管对细菌脂多糖输注的渗透性增加。数据显示，脂多糖增加了微静脉和毛细血管段的液体渗漏。因此，该方法可以比较血管段之间的渗透性响应，从而定义响应中的任何异质性。虽然通常使用的定义肺渗透性的方法需要对肺组织样品进行后处理，但是使用真实的时间成像消除了这种要求，这从本方法中是显而易见的。因此，与真实的时间成像相结合的分离的肺准备提供了优于传统方法的几个优点来确定肺微血管渗透性，但仍然是开发和实施的直接方法。
The isolated blood-perfused lung preparation is widely used to visualize and define signaling in single microvessels. By coupling this preparation with real time imaging, it becomes feasible to determine permeability changes in individual pulmonary microvessels. Herein we describe steps to isolate rat lungs and perfuse them with autologous blood. Then, we outline steps to infuse fluorophores or agents via a microcatheter into a small lung region. Using these procedures described, we determined permeability increases in rat lung microvessels in response to infusions of bacterial lipopolysaccharide. The data revealed that lipopolysaccharide increased fluid leak across both venular and capillary microvessel segments. Thus, this method makes it possible to compare permeability responses among vascular segments and thus, define any heterogeneity in the response. While commonly used methods to define lung permeability require postprocessing of lung tissue samples, the use of real time imaging obviates this requirement as evident from the present method. Thus, the isolated lung preparation combined with real time imaging offers several advantages over traditional methods to determine lung microvascular permeability, yet is a straightforward method to develop and implement.