QUANTITATIVE TWO PHOTON EXCITED CALCIUM UNCAGING

一种测量双光子去势作用的技术 本实验室已研制出钙笼横截面。 $^{1，2}$。使用这项技术，我们报告了对 新型钙笼DMNPE-4的有效双光子激发 $^{3}$。我们还描述了预测总金额的计算 未被释放的钙离子和被释放的人的时间行为 给定多光子光解条件下的钙分布 用一个给定的笼子。这些计算与双光子相结合 我们已经测量了各种不同的 钙笼可用于钙依赖的定量研究 细胞或厚组织内具有高空间分辨率的突起 样本。这些研究的最终目的是应用 这些工具可用于自发瞬变的定量研究 骨骼发育过程中钙浓度升高(“火花”) 肌细胞，探索增强/抑制关系 在一个局部区域中的连续火花之间以及在 在空间上不同的区域产生火花。1.谢尔，J.等人。{\em 生物群落。70(2)A211(1996)2.Brown等人，手稿#年 准备(1997)3.Ellis-Davies，G.J.)72(2)A246 (1997年)

A technique for the measurement of the two-photon uncaging action cross section of calcium cages has been developed in this laboratory $^{1,2}$. Using this technique, we report on the prognosis for effective two-photon excitation of a novel calcium cage, DMNPE-4 $^{3}$. We also describe calculations which predict the total amount of calcium ions uncaged and the temporal behavior of the liberated calcium distribution under given multiphoton photolysis conditions with a given cage. These calculations, combined with the two-photon uncaging action cross sections that we have measured for a variety of calcium cages enable the quantitative study of calcium dependent processes with high spatial resolution within cells or thick tissue samples. The ultimate intent of these studies is the application of these tools to the quantitative study of the spontaneous transient calcium concentration elevations ("sparks") in developing skeletal muscle cells, probing the potentiation/inhibition relation ships between consecutive sparks in one localized region as well as between sparks in spatially separate regions. 1. Shear, J. et al. {\em Biophys. J.}70(2) A211 (1996) 2. Brown et al, Manuscript in Preparation (1997) 3. Ellis-Davies, G.{\em Biophys. J.}72(2) A246 (1997)