Chen Yun, Xiong Guohua, Arriaga Edgar A
Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
Electrophoresis. 2007 Jul;28(14):2406-15. doi: 10.1002/elps.200600628.
The properties of organelles within a cell have been shown to be highly heterogeneous. Until now, it has been unclear just how much of this heterogeneity is endemic to the organelle subpopulations themselves and how much is actually due to stochastic cellular noise. An attractive approach for investigating the origins of heterogeneity among the organelles of a single cell is CE with LIF detection (CE-LIF). As a proof of principle, in this report we optimize and use a single cell CE-LIF method to investigate the properties of endocytic (acidic) organelles. Our results show that the properties of individual acidic organelles containing Alexa Fluor 488 Dextran suggest that there are two groups of CCRF-CEM cells: a group with a high dextran content per cell, and a group with a low dextran content per cell. Furthermore, the individual organelle measurements of the single cells allow us to compare in each group the distributions of doxorubicin content per acidic organelle and electrophoretic mobilities of these organelles.
细胞内细胞器的特性已被证明具有高度异质性。到目前为止,尚不清楚这种异质性在细胞器亚群本身中占多大比例,以及实际上有多少是由于随机细胞噪声所致。一种用于研究单个细胞内细胞器异质性起源的有吸引力的方法是采用激光诱导荧光检测的毛细管电泳(CE-LIF)。作为原理验证,在本报告中,我们优化并使用单细胞CE-LIF方法来研究内吞(酸性)细胞器的特性。我们的结果表明,含有Alexa Fluor 488葡聚糖的单个酸性细胞器的特性表明存在两组CCRF-CEM细胞:一组细胞内葡聚糖含量高,另一组细胞内葡聚糖含量低。此外,对单个细胞的细胞器进行单独测量,使我们能够在每组中比较每个酸性细胞器中阿霉素含量的分布以及这些细胞器的电泳迁移率。