Confocal imaging to quantify passive transport across biomimetic lipid membranes.
Confocal imaging to quantify passive transport across biomimetic lipid membranes.
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DOI：
10.1021/ac1016826
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发表时间：
2010-09-15
影响因子：
7.4
通讯作者：
Malmstadt, Noah
中科院分区：
文献类型：
--
作者：
Li, Su;Hu, Peichi;Malmstadt, Noah
分子在没有任何活性细胞机制的帮助下通过质膜的能力是该分子的药物特性的核心。被动转运已经在奥弗顿规则的背景下被理解，该规则指出，更多的亲脂分子更容易穿过膜脂双层。现有的测量被动输运的技术缺乏可重复性，并且受到未搅拌层（USL）的存在的阻碍，该层主导着双分子层的输运。本报告描述了基于旋转盘共聚焦显微镜（SDCM）的巨型单层囊泡（guv）的分析，可以详细研究被动运输过程和机制。这种方法可以直接观察浓度场，从而可以很容易地从瞬态浓度剖面数据中确定膜的渗透率。构建了一系列亲水性增强的分子，并观察了这些分子在guv中的转运情况。观察到的渗透率趋势符合Overton规则。然而，测量值偏离了被动输运的简单分区-扩散比例模型。这项技术很容易实现，作为一种测量膜运输的方法有很大的希望。它最适合于被动输运对膜性质依赖的精确定量测量。
The ability of a molecule to pass through the plasma membrane without the aid of any active cellular mechanisms is central to that molecule’s pharmaceutical characteristics. Passive transport has been understood in the context of Overton’s rule, which states that more lipophilic molecules cross membrane lipid bilayers more readily. Existing techniques for measuring passive transport lack reproducibility and are hampered by the presence of an unstirred layer (USL) that dominates transport across the bilayer. This report describes assays based on spinning-disk confocal microscopy (SDCM) of giant unilamellar vesicles (GUVs) that allow for the detailed investigation of passive transport processes and mechanisms. This approach allows the concentration field to be directly observed, allowing membrane permeability to be determined easily from the transient concentration profile data. A series of molecules of increasing hydrophilicity was constructed and the transport of these molecules into GUVs was observed. The observed permeability trend is consistent with Overton’s rule. However, the values measured depart from the simple partition-diffusion proportionality model of passive transport. This technique is easy to implement and has great promise as an approach to measure membrane transport. It is optimally suited to precise quantitative measurements of the dependence of passive transport on membrane properties.
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