Bootman Martin D, Rietdorf Katja, Collins Tony, Walker Simon, Sanderson Michael
Babraham Institute, Babraham, Cambridge, CB22 3AT, United Kingdom.
Cold Spring Harb Protoc. 2013 Feb 1;2013(2):83-99. doi: 10.1101/pdb.top066050.
Imaging Ca(2+)-sensitive fluorescent indicators provides a common approach for studying Ca(2+) signals in many contexts. Fluorescent indicators are particularly useful for measuring acute Ca(2+) changes in a relatively noninvasive manner. The availability of indicators that can be targeted to specific cellular domains, coupled with variations in affinity, brightness or spectral characteristics, provides tools for exploring spatially and temporally diverse Ca(2+) signals, and moreover, multiplexing the readout of Ca(2+) with other cellular functions. This article aims to give the novice experimentalist some insight into the considerations and potential pitfalls that impinge on the use of fluorescent Ca(2+) indicators.
对Ca(2+)敏感的荧光指示剂成像为在多种情况下研究Ca(2+)信号提供了一种常用方法。荧光指示剂对于以相对非侵入性的方式测量急性Ca(2+)变化特别有用。能够靶向特定细胞区域的指示剂的可用性,再加上亲和力、亮度或光谱特性的变化,为探索空间和时间上多样的Ca(2+)信号提供了工具,此外,还能将Ca(2+)的读数与其他细胞功能进行多重分析。本文旨在让新手实验人员对影响荧光Ca(2+)指示剂使用的注意事项和潜在陷阱有所了解。