Dominak Lisa M, Keating Christine D
Department of Chemistry, Pennsylvania State University, University Park, PA 16802, USA.
Langmuir. 2007 Jun 19;23(13):7148-54. doi: 10.1021/la063687v. Epub 2007 May 22.
We report encapsulation of polymers and small molecules within individual giant lipid vesicles (GVs; 3-80 microm), as determined by confocal fluorescence microscopy. Polymer-bound or free dyes were encapsulated within GVs by including these molecules in the aqueous solution during vesicle formation via gentle hydration. Encapsulation efficiencies of individual GVs (EE(ind)) were determined from the fluorescence intensity ratio inside vs outside the vesicle. EE(ind) varied considerably from vesicle to vesicle, with interior solute concentrations for GVs within the same batch ranging from much less than to slightly more than the initial concentration. The majority of GVs had high internal concentrations of polymer or small-molecule encapsulants equal to or slightly greater than the external concentration. EE(ind) decreased for high molecular weight polymers (e.g., dextran 500 000), but was relatively insensitive to the GV diameter, membrane composition, or incubation temperature in our experiments. Knowledge of EE(ind) is important for quantitative evaluation of reactions occurring within GVs (e.g., enzymatic processes) and for optimizing encapsulation conditions.
我们报告了通过共聚焦荧光显微镜测定,聚合物和小分子被包裹在单个巨型脂质囊泡(GVs;3 - 80微米)内的情况。在通过温和水合作用形成囊泡的过程中,将聚合物结合的染料或游离染料包含在水溶液中,从而将其包裹在GVs内。单个GVs的包封效率(EE(ind))由囊泡内部与外部的荧光强度比来确定。EE(ind)在不同囊泡之间有很大差异,同一批次内GVs的内部溶质浓度范围从远低于初始浓度到略高于初始浓度。大多数GVs具有高分子或小分子封装剂的高内部浓度,等于或略大于外部浓度。在我们的实验中,对于高分子量聚合物(例如葡聚糖500 000),EE(ind)会降低,但对GV直径、膜组成或孵育温度相对不敏感。了解EE(ind)对于定量评估GVs内发生的反应(例如酶促过程)以及优化包封条件很重要。