光电结合检测大鼠原代神经元胞内钙离子浓度方法的建立

A Novel Method for Recording Intracellular Calcium Ion Concentration of Primary Neurons Using Confocal Microscopy System Combined with Electrical Stimulation

目的利用电刺激结合转盘共聚焦显微镜检测胞内钙离子浓度变化。方法钙离子变化是神经元发挥功能的关键因素。本实验利用荧光探针fluo-4AM标记钙离子,电刺激兴奋神经元,转盘共聚焦技术检测胞内荧光强度的快速变化。结果成功制备了刺激电极,并结合转盘共聚焦系统检测到刺激前10%左右的原代神经元有自发的钙离子升高,电刺激后,胞内钙离子迅速上升,刺激结束后,仍有周期性的升高,但幅度降低。结论电刺激结合转盘共聚焦检测到胞内刺激前后钙离子的快速变化。

Objective To detect changes in intracellular calcium ion using a detection system of spinning disk confocal microscopy combined with electrical stimulation. Method Calcium ion change is one of the key factors for neuron function. In this experiment,the fluorescent probe fluo-4 AM was used to label calcium ions,neurons were excited by electrode stimuli and the spinning disk confocal was used to detect the rapid changes of intracellular fluorescence intensity. Result The electrode was successfully prepared. It was detected that about 10% of the primary neurons had spontaneous calcium ion rise. After electrical stimulation,the intracellular calcium ion rose rapidly. When the stimulation ended,the calcium ion still rise,but the amplitude decreased. Conclusion Spinning disk confocal microscopy combined with electrical stimulation system detects rapid changes of calcium ions before and after stimulation.