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应用ACAS570激光扫描共聚焦显微镜（laserscanningconfocalmicroscope，LSCM简称共焦显微镜）和钙离子指示剂Fluo－3／AM、FuraRed荧光探剂双标记技术，测定了单个活细胞内比率钙的动态变化。结果显示37℃，Fluo－3／AM浓度10μmol／L，FuraRed浓度10μmol／L的条件下，昆明小鼠巨噬细胞负载1h左右即可获良好的标记效果。用比率探针测量细胞内Ca~2＋的动态变化，使Ca~2＋的定性定量测定不受染料浓度、细胞大小、照射光强度以及一定程度的光漂白和染料泄漏等因素的影响，提供了一种准确定性定量细胞内Ca~2＋的实时、动态、原位变化的新方法。
The dynamic changes of calcium in a single living cell were measured by ACAS570 laser scanning confocal microscopy (LSCM) and Fluo-3 / AM with calcium ion indicator and FuraRed fluorescent double labeling technique. The results showed that under the conditions of 37 ℃, Fluo-3 / AM concentration of 10μmol / L and FuraRed concentration of 10μmol / L, the macrophages of Kunming mice could be well labeled for about 1h. Using the ratio probe to measure the intracellular Ca2 + dynamic changes, the qualitative and quantitative determination of Ca2 + is not affected by the concentration of the dye, the size of the cell, the intensity of the irradiated light and a certain degree of photo-bleaching and dye leakage, and provides a A New Method to Accurately Qualify Quantitative and Quantitative Intracellular Ca ~ (2 +) Changes in Real Time, Dynamic and In Situ.