传统与光谱流式细胞仪对自发荧光样本检测的比较

Comparative investigation for auto-fluorescence sample detection by evaluating conventional and spectral flow cytometers performance

目的比较传统与光谱流式细胞仪检测自发荧光样本的性能。方法以annexin V(AV)/碘化丙啶(PI)染色检测细胞凋亡为模型,制备带有自发荧光的单细胞样本,分别用传统与光谱流式细胞仪采集同一样本荧光信号。结果自发荧光干扰异硫氰酸荧光素(FITC)通道荧光信号,传统流式细胞仪应避免使用FITC通道的荧光素;而光谱流式细胞仪可以清楚分辨FTIC通道的荧光素和自发荧光。同时优化了样本制备方法和annexin V/PI染色方案。结论光谱流式细胞仪对于自发荧光样本的检测在一定条件下具有相对优良的性能。

Objective To compare the performance of conventional and spectral flow cytometers in detecting auto-fluorescence samples.Methods Single cell samples with auto-fluorescence were prepared and stained by annexin V(AV)binding and propidium iodure(PI)to detect apoptosis.The same auto-fluorescence sample was run on conventional and spectral flow cytometers side by side.Results By avoid using FITC in conventional cytometers,the better result was gotten in detecting auto-fluorescence samples.Full spectrum cytometry presented better resolution even there was spectral overlap.It was easy to discriminate the signals derived from auto-fluorescence and FITC.The protocol of annexinV binding and PI staining to analyze apoptosis was optimized.Conclusions Spectral flow cytometry shows relatively excellent performance under certain conditions in detecting of apoptosis derived from auto-fluorescence samples.