Goldberg G S, Bechberger J F, Naus C C
University of Western Ontario, London, Canada.
Biotechniques. 1995 Mar;18(3):490-7.
We describe a simple method for evaluating gap junctional communication (GJC) between cells in culture. The procedure involves pre-loading cells with two fluorescent dyes: calcein and DiI. Calcein is able to pass through gap junctions, while DiI is not. These pre-loaded cells are then plated with unlabeled cells. The number of cells receiving calcein from each pre-loaded cell can then be quantified after the cells settle on the plate. Potent and reversible inhibitors of GJC can be used in this system to evaluate dye transfer within a given period of time.
我们描述了一种评估培养细胞间间隙连接通讯（GJC）的简单方法。该程序包括用两种荧光染料：钙黄绿素和DiI对细胞进行预加载。钙黄绿素能够通过间隙连接，而DiI则不能。然后将这些预加载的细胞与未标记的细胞一起接种。在细胞在平板上沉降后，可以对从每个预加载细胞接收钙黄绿素的细胞数量进行量化。GJC的强效和可逆抑制剂可用于该系统，以评估给定时间段内的染料转移。
