成像流式细胞术和免疫荧光技术检测成纤维样滑膜细胞Ahr入核的比较

Comparison of imaging flow cytometry and immunofluorescence techniques for detection of Ahr entry in fibroblast-like synoviocytes

目的探究成像流式细胞术和免疫荧光技术检测成纤维样滑膜细胞中Ahr入核的差别。方法用15%的DMEM培养人来源的成纤维样滑膜细胞细胞株MH7A,分别用激光共聚焦显微镜和量化成像流式细胞仪(ImageStreamX MarkⅡ)检测空白对照组、Kyn组和Kyn+CH223191组对MH7A中Ahr的入核水平,并采用非参数检验比较两种检测技术检测结果的相关性。结果与空白对照组相比,免疫荧光技术和成像流式细胞术测得的MH7A细胞中Kyn组和Kyn+CH223191组Ahr入核能力增加(P<0.05),两种实验方法测得的三组结果相关性良好,R 2值分别是0.8638、0.9287和0.9018,差异有统计学意义(P<0.05)。结论相较于免疫荧光技术的操作和结果,成像流式细胞术数据处理比较复杂,但所得结果精确度高,避免了实验者的主观性,减少实验误差。

Objective To investigate the difference between imaging flow cytometry and immunofluorescence techniques for detecting Ahr entry in fibroblast-like synoviocytes.Methods Human-derived fibroblast-like synoviocytes line MH7A was cultured with 15%DMEM.The nucleation levels of Ahr in MH7A were detected by laser scanning confocal microscopy and quantitative imaging flow cytometry(ImageStreamX Mark II)in the blank control group,the Kyn group,and the Kyn+CH223191 group,respectively.The results of the two assay techniques were compared using nonparametric tests-correlation of the results.Results Compared with the blank control group,the Kyn and Kyn+CH223191 groups of MH7A cells increased Ahr nucleation ability as measured by immunofluorescence technique and imaging flow cytometry(P<0.05),and the three groups of results calculated by the two experimental methods were consistent,with the R2 values of 0.8638,0.9287,and 0.9018,respectively,which were statistically significant differences(P<0.05).Conclusion Compared with the operation and results of immunofluorescence experiments,imaging flow cytometry data processing is more complicated.However,the results are notably precise,avoiding the subjectivity of the experimenter and minimizing experimental inaccuracies.