基于荧光时延估计的流式细胞仪荧光信号表征方法

摘要

针对流式细胞仪荧光信号强度测量过程存在非线性,导致细胞分群分析结果不准确的问题,提出了一种基于荧光时延估计的流式细胞仪荧光信号表征方法代替荧光寿命测量,实现对荧光脉冲信号的表征.首先,介绍了荧光脉冲产生及时延的原理;然后,对荧光信号强度表达式进行推演,得到荧光时延表征荧光寿命的结论;最后,利用高斯拟合对该荧光信号表征方法进行了实验验证.实验结果表明,相对于荧光脉冲强度表征方法,荧光脉冲时延估计表征方法具有更小的分布范围,更小的标准差.限定于本实验系统,单色荧光脉冲信号的时延估计表征方法可将峰值表征方法的标准差由0.118降至0.007.%Aiming at the problem that nonlinear existed in the measurement process of fluorescence signal intensity of flow cytometer,which led to the inaccurate of results of cell clustering analysis,a fluorescence time delay estimation method was pro-posed to replace the fluorescence lifetime measurement.First of all,the principle of fluorescence pulse generation and time delay were introduced.Then,the expression of fluorescence signal intensity was deduced and the conclusion was obtained that the fluo-rescence lifetime was characterized by fluorescence delay.Finally,the fluorescence signal characterization method was validated by Gaussian fitting.The experiment results indicate that the method of fluorescence pulse time delay estimation has a smaller distribu-tion range and a smaller standard deviation compared with the fluorescence intensity characterization method. In this experiment system,the standard deviation of peak characterization method reduces from 0. 118 to 0. 007 based on the time delay estimation method of monochromatic fluorescence pulse signal.