基于EnVision多标记读板仪的细胞内钙信号检测技术的研究

Study on measurement of intracellular calcium signaling based on En Vision multilabel plate reader

目的研究使用En Vision多标记读板仪检测细胞内钙信号的可行性。方法分别用卡巴胆碱和凝血酶刺激LN229、SKN-MC细胞或者磷脂酶C抑制剂U73122预处理的LN229细胞,使用En Vision多标记读板仪检测细胞内钙信号强度的变化。结果卡巴胆碱诱导的LN229细胞内钙信号变化呈现浓度依赖性;卡巴胆碱和凝血酶诱导的细胞内钙信号变化趋势不同;凝血酶诱导的钙信号变化具有细胞特异性;U73122对凝血酶诱导的钙信号的抑制作用呈现剂量依赖性。结论 En Vision多标记读板仪作为细胞内钙信号检测仪器,具有快速、可重复性好和灵敏度高等优势,适用于钙信号相关的研究和小分子抑制剂的筛选等领域的工作。

Objective To investigate the feasibility of measuring the intracellular calcium signaling using EnVision muhilabel plate reader. Methods LN229 and SKN-MC cells were stimulated with carbachol and thrombin, respectively. LN229 cells were pretreated with PLC inhibitor U73122 and then stimulated with thrombin. Intracellular calcium signaling was measured using EnVision multilabel plate reader. Results Carbachol induced the calcium signaling in LN229 cells in a concentration-dependent manner. Carbachol and thrombin-induced calcium signaling exhibited different profile. Thrombin-induced calcium signaling was dependent on cell type. U73122 inhibited thrombin-stimulated calcium signaling in a dose-dependent manner. Conclusion EnVision muhilabel plate reader can be utilized for detecting intracellular calcium signaling, which is rapid, highly repeatable and sensitive.