流式细胞技术检测细胞内雌激素受体方法的探讨

Investigations into the method that estrogen receptor content in cell is assayed by flow cytometry

目的 建立一种灵敏度高、特异性强的细胞内雌激素受体检测方法。方法 通过加入不同的试剂浓度及改变细胞渗透剂等 ,观察细胞内雌激素受体的表达。结果 随加入抗体浓度的增加 ,细胞内雌激素受体表达增加 ,用 0 5 %Saponin溶液穿透细胞膜较用 0 1%Tritonx 10 0 ,雌激素受体表达水平明显为高 (P <0 0 1)。结论 在用流式细胞技术检测细胞内雌激素受体时 ,加入抗体的最佳浓度为 45 μg/ml;细胞渗透剂以用 0 5 %Saponin溶液为宜。

Objective: To find a highly sensitive and specific method by which estrogen receptor in cells can be assayed. Methods: The level of estrogen receptor was estimated by varying antibody concentrations and different cell membrance permeability agents. Results: Within given antibody concentration range, both specific fluorescence and total fluorescence significantly increased as a result of added antibody concentration increase. The expression level of estrogen receptor with 0.5% Saponin permeability was greatly higher than that with 0.1% Tritonx-100. Statistical analysas show significant differences(P<001). Conclution: In the flow cytometry by which the estrogen receptor in cells is assayed, the optimal antibody concentration is 45 μg/ml, and 0.5% Saponin in permeating cell membrane is appropriate.