Sato Kanetomo, Obinata Kei, Sugawara Tadashi, Urabe Itaru, Yomo Tetsuya
Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, 2-1 Yamadaoka, Suita, Osaka, Japan.
J Biosci Bioeng. 2006 Sep;102(3):171-8. doi: 10.1263/jbb.102.171.
We describe a new high-throughput method of quantifying the structural properties of individual cell-sized liposomes. An internal aqueous solution of liposomes was labeled with a green fluorescent marker and the membrane with a red marker. The double-labeled liposomes were analyzed using flow cytometry, and the internal aqueous volume and lipid membrane volume of each liposome were measured. The experimental results indicate that both the internal aqueous and lipid membrane volumes positively correlate with the intensity of forward-scatter (FS) and side-scatter (SS) signals in a logarithmic scale. In addition, liposomes in 18 small areas gated by log(FS) and log(SS) were sorted by fluorescence-activated cell sorting (FACS), and observed by optical microscopy. Structural characteristics observed in the microscopy images of heterogeneous liposomes correlated with FACS data. Because this method does not employ any particular assumption about the shape and structure of liposomes, flow cytometry is a powerful tool for estimating the internal and membrane volumes of individual cell-sized liposomes with heterogeneous shapes and structures.
我们描述了一种用于量化单个细胞大小脂质体结构特性的新的高通量方法。脂质体的内部水溶液用绿色荧光标记物标记，膜用红色标记物标记。使用流式细胞术分析双标记脂质体，并测量每个脂质体的内部水相体积和脂质膜体积。实验结果表明，内部水相和脂质膜体积在对数尺度上均与前向散射（FS）和侧向散射（SS）信号强度呈正相关。此外，通过对数（FS）和对数（SS）门控的18个小区域中的脂质体通过荧光激活细胞分选（FACS）进行分选，并通过光学显微镜观察。在异质脂质体的显微镜图像中观察到的结构特征与FACS数据相关。由于该方法不对脂质体的形状和结构做任何特定假设，流式细胞术是估计具有异质形状和结构的单个细胞大小脂质体的内部和膜体积的有力工具。
