Pozharski Edwin V, Macdonald Robert C
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, IL 60208, USA.
Anal Biochem. 2005 Jun 15;341(2):230-40. doi: 10.1016/j.ab.2005.03.027.
A flow fluorometric approach to study cationic lipoid-DNA complexes is presented. The approach uses standard flow cytometry equipment and common fluorescent dyes (BODIPY and ethidium homodimer-2) to detect both lipoid and DNA content in individual particles. In addition, a procedure that allows determination of whether or not liposomes remain intact is described. The procedure is based on monitoring the retention of a polar tracer that has been preloaded into its aqueous compartment. Sample preparation, instrument setup, data analysis, and methodological limitations are described. Applications of the procedure to cationic lipoid-DNA complexes are described, and illustrations are given for the determination of how the lipoid content, composition, and structure of individual lipoplexes in a population evolve over time, starting at about 1 min after DNA and vesicles are mixed. Analogous procedures can be applied to other heterogeneous particles and supramolecular structures.
本文介绍了一种用于研究阳离子脂质体 - DNA 复合物的流动荧光测定方法。该方法使用标准流式细胞仪设备和常见荧光染料（BODIPY 和乙锭同二聚体 -2）来检测单个颗粒中的脂质体和 DNA 含量。此外，还描述了一种可用于确定脂质体是否保持完整的程序。该程序基于监测预先加载到其水相隔室中的极性示踪剂的保留情况。文中描述了样品制备、仪器设置、数据分析以及方法学局限性。阐述了该程序在阳离子脂质体 - DNA 复合物中的应用，并给出了示例，说明从 DNA 和脂质体混合后约 1 分钟开始，群体中单个脂质体复合物的脂质体含量、组成和结构如何随时间演变。类似的程序可应用于其他异质颗粒和超分子结构。
