Messerli Mark A, Smith Peter J S
BioCurrents Research Center, Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, Massachusetts, USA.
Methods Cell Biol. 2010;99:91-111. doi: 10.1016/B978-0-12-374841-6.00004-9.
Ca(2+) signaling in the extra- and intracellular domains is linked to Ca(2+) transport across the plasma membrane. Noninvasive monitoring of these resulting extracellular Ca(2+) gradients with self-referencing of Ca(2+)-selective microelectrodes is used for studying Ca(2+) signaling across Kingdoms. The quantitated Ca(2+) flux enables comparison with changes to intracellular [Ca(2+)] measured with other methods and determination of Ca(2+) transport stoichiometry. Here, we review the construction of Ca(2+)-selective microelectrodes, their physical characteristics, and their use in self-referencing mode to calculate Ca(2+) flux. We also discuss potential complications when using them to measure Ca(2+) gradients near the boundary layers of single cells and tissues.
细胞外和细胞内区域的Ca(2+)信号传导与Ca(2+)跨质膜运输相关联。利用Ca(2+)选择性微电极的自参考对这些产生的细胞外Ca(2+)梯度进行无创监测，用于研究跨生物界的Ca(2+)信号传导。定量的Ca(2+)通量能够与用其他方法测量的细胞内[Ca(2+)]变化进行比较，并确定Ca(2+)运输化学计量。在此，我们综述了Ca(2+)选择性微电极的构建、其物理特性以及它们在自参考模式下用于计算Ca(2+)通量的应用。我们还讨论了在使用它们测量单细胞和组织边界层附近的Ca(2+)梯度时可能出现的并发症。
