Bootman Martin D, Roderick H Llewelyn
Laboratory of Molecular Signalling, Babraham Institute, Cambridge, UK.
Methods Mol Biol. 2011;746:277-96. doi: 10.1007/978-1-61779-126-0_15.
Monitoring cellular calcium concentration using fluorescent reporters can provide a rapid, proportional assay of G-protein-coupled receptor activation. Recording calcium changes in single cells, or cell populations, is relatively straightforward, but requires careful deliberation regarding the appropriate calcium reporter and experimental approach. Here, we describe strategies to ensure that calcium changes are recorded with good fidelity and minimal invasiveness. We highlight a range of issues that need to be considered within the design of an experiment to measure cellular calcium, and suggest strategies to avoid common pit-falls.
使用荧光报告基因监测细胞钙浓度能够提供一种快速、成比例的G蛋白偶联受体激活检测方法。记录单个细胞或细胞群体中的钙变化相对简单,但需要仔细考虑选择合适的钙报告基因和实验方法。在此,我们描述了确保以高保真度和最小侵入性记录钙变化的策略。我们强调了在设计测量细胞钙的实验时需要考虑的一系列问题,并提出了避免常见陷阱的策略。