Vorauer-Uhl K, Wagner A, Borth N, Katinger H
Institute of Applied Microbiology, University of Agricultural Sciences, Vienna, Austria.
Cytometry. 2000 Feb 1;39(2):166-71.
An essential parameter that describes the quality of liposome suspensions is the mean size, respectively the size distribution. Currently several analytical methods including laser light scattering techniques (LLST) are being employed.
Here we present an alternative technique using flow cytometry (FCM) to characterize uni- and polydisperse suspensions. As model liposomes preparations containing dipalmitoylphosphatidylcholine (DPPC) were used. A constant number of particles (1,500/s) in the fluid stream and a representative number of 10,000 particles of each sample was measured. Fluorescence-labeled latex beads were measured identically, and their side scatter signals were calibrated and correlated to the results obtained with liposome vesicles.
Evaluation of the measurement and validation of the FCM results in comparison to LLST confirm the reliability of results obtained with our method. Latex beads in the range of 100-1000 nm were used for calibration to classify liposomes. Although measurement characteristics and calculation in both methods are basically different, very good agreement of the results was achieved.
Demonstration of stability, reproducibility, and reliability of results make the employment of this method acceptable for an adequate routine analysis technique.
描述脂质体悬浮液质量的一个重要参数是平均粒径，即粒径分布。目前正在使用包括激光散射技术（LLST）在内的几种分析方法。
在此，我们提出一种使用流式细胞术（FCM）来表征单分散和多分散悬浮液的替代技术。使用含有二棕榈酰磷脂酰胆碱（DPPC）的脂质体制剂作为模型。测量流体流中恒定数量的颗粒（1500个/秒），并对每个样品测量10000个代表性颗粒。对荧光标记的乳胶珠进行相同的测量，并校准其侧向散射信号，并将其与脂质体囊泡获得的结果相关联。
与LLST相比，对测量结果的评估和FCM结果的验证证实了我们方法获得的结果的可靠性。使用100 - 1000nm范围内的乳胶珠进行校准以对脂质体进行分类。尽管两种方法的测量特性和计算方法基本不同，但结果非常吻合。
结果的稳定性、可重复性和可靠性证明使得该方法适用于适当的常规分析技术。
