Smith Godfrey, Reynolds Martyn, Burton Francis, Kemi Ole Johan
School of Life Sciences, University of Glasgow, United Kingdom.
Methods Cell Biol. 2010;99:225-61. doi: 10.1016/B978-0-12-374841-6.00009-8.
This chapter compares the imaging capabilities of a range of systems including multiphoton microscopy in regard to measurements of intracellular Ca(2+) within living cells. In particular, the excitation spectra of popular fluorescent Ca(2+) indicators are shown during 1P and 2P excitation. The strengths and limitations of the current indicators are discussed along with error analysis which highlights the value of matching the Ca(2+) affinity of the dye to a particular aspect of Ca(2+) signaling. Finally, the combined emission spectra of Ca(2+) and voltage sensitive dyes are compared to allow the choice of the optimum combination to allow simultaneous intracellular Ca(2+) and membrane voltage measurement.
本章比较了一系列系统的成像能力，包括多光子显微镜在活细胞内钙离子（Ca(2+)）测量方面的能力。特别地，展示了常用荧光钙指示剂在单光子（1P）和双光子（2P）激发下的激发光谱。讨论了当前指示剂的优缺点，并进行了误差分析，突出了使染料的钙亲和力与钙信号传导的特定方面相匹配的价值。最后，比较了钙离子和电压敏感染料的组合发射光谱，以便选择最佳组合来同时测量细胞内钙离子和膜电压。
