单个牛视网膜神经细胞内游离Ca2+动态变化的实时观测方法

为观测神经营养因子对胞内Ca2+浓度的影响，将牛视网膜神经细胞内的Ca2+用Fluo-3标记，用搭建的活细胞实时成像荧光显微系统对其成像，并观测在脑源性神经营养因子BDNF等四种神经营养因子的作用下细胞内游离Ca2+浓度([Ca2+]_i)随时间的变化的规律。由于荧光分子有自发衰减效应，故对得到的细胞内荧光强度采用“去衰减效应修正”的处理方法，再现了较真实地代表[Ca2+]_i的荧光强度。修正后的数据显示，加入四种神经因子后，[Ca2+]_i皆有不同程度的升高，与相关文献所描述的类似实验具有相似结果，说明此种对活细胞内荧光标记物的实时动态成像的实验方法可行，去衰减效应修正的数据处理方法可靠。

A Method for Real-Time Observation of Intracellular Free Ca2+ Concentration in Single Bovine Retinal Neurons

In order to observe the neurotrophin’s influence on intracellular free calcium concentration, the test neuron was labeled by fluorescence indicator Fluo-3, and imaged by self-built real-time fluorescence microscopy system. The authors observed the changes in intracellular free calcium concentration ([Ca2+]_i) in the bovine retinal neurons under the influence of four kinds of neurotrophins such as brain derived neurotrophic factor BDNF. On account of the fact that fluorescence indicator’s intensity decays over time, it is necessary to apply a “decay removal correction” to the fluorescence intensity in order to show the fluorescence intensity that solely represents [Ca2+]_i. The corrected data shows an increase after adding neurotrophins to neurons, which is consistent with similar results published by other groups. Thus, our method of imaging living cells is feasible, and “decay removal correction” is reliable.