荧光探针Fluo-3/AM和FuraRed测量单细胞中比率钙  

Ratiometric calcium measurements in single cells with visible wavelength probes fluo-3/AM and furared

作　　者：马晓冬[1] 朱晓亮[2] 赵清[1] 张瑾[1] 张飒[1] 雷国华[1] 黄行许[1] 鲍永耀[1] 

机构地区：[1]第一军医大学中心实验室,广东广州510515 [2]南方医院皮肤科,广东广州510515

出　　处：《中国医学物理学杂志》1999年第3期180-182,共3页Chinese Journal of Medical Physics

摘　　要：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。Using laser scanning confocal microscope (LSCM)and highly sensitive Ca^2+ fluorescent dye, Fluo-3/AM and FuraRed, we measureed the kinetic changes of ratiometric calcium in single intact living cells. The re-sults showed, incubated for 1h at 37℃ with 10 μmol/L Fluo-3/AM and 10 μmol/L FuraRed, the murine peri-toneal macrophages cultured on gass cover slips were manifested excellent LSCM imaging of intracellular Ca^2+.The use of ratiometric probes can be qualified and quantified intracelluIar Ca^2+ more accurately, since it allowscalcium quantitation independent of dye concentration - cell volume, illumination intensity, and reduces the effectof photobleaching and dye leakage. The results indicated that, by using laser scanning confocal microscope andsensitive Ca^2+ fluorescent dye , Flul- 3/AM and FuraRed,provided a exactly new method for real time continuousand in situ detection of kinetic variations in intracellular Ca^2+.

关 键 词：激光扫描 共聚焦显微镜 比率钙 荧光探针 

分 类 号：Q582[生物学—生物化学] Q26