Automated, high-resolution micropipet aspiration reveals new insight into the physical properties of fluid membranes
Automated, high-resolution micropipet aspiration reveals new insight into the physical properties of fluid membranes
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DOI：
10.1021/la047801q
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发表时间：
2005-03-01
期刊：
影响因子：
3.9
通讯作者：
Rawicz, W
中科院分区：
文献类型：
--
作者：
Heinrich, V;Rawicz, W
我们描述了最近的进展，我们的实验方法来检查生物和人工膜的物理特性，自动化微量吸管抽吸。新的仪器使我们能够应用快速而精确的张力协议的膜，同时连续记录膜变形与高速视频显微镜。亚像素几何测量的鲁棒算法跟踪膜边缘的位移，分辨率为几个纳米，使我们能够确定膜面积的变化和封闭体积的吸移管抽吸细胞或囊泡异常准确。实验和数据分析大大方便了定制编写的软件，其基本设计也在这里描述。示例测量演示了这种技术如何显着提高了通过各种类型的微量吸移管抽吸实验获得的数据的量和可靠性，使我们能够研究膜行为的有趣方面，这些方面都回避了早期的技术。
We describe recent advances in our experimental approach to examine the physical properties of biological and artificial membranes by automated micropipet aspiration. New instrumentation allows us to apply fast yet precise tension protocols to membranes while continuously recording the membrane deformation with high-speed videomicroscopy. Robust algorithms for subpixel geometric measurements track the displacements of membrane edges with resolution of a few nanometers and enable us to determine changes of the membrane area and enclosed volume of pipet-aspirated cells or vesicles with exceptional accuracy. Experimentation and data analysis are greatly facilitated by custom-written software whose basic design is described here as well. Example measurements demonstrate how this technique has significantly improved the amount and reliability of data obtained by various types of micropipet-aspiration experiments, allowing us to study interesting aspects of membrane behaviour that have eluded earlier techniques.