流式细胞计数法评定不同抗凝剂对骨髓样本红细胞的裂解作用

Evaluation of different anticoagulants in cracking myeloid erythrocytes by flow cytometry

目的初步探讨临床常用的肝素钠、乙二胺四乙酸盐(EDTA)、草酸盐等抗凝剂在流式细胞仪细胞采集、分析过程中,对骨髓样本红细胞裂解作用的影响。方法收集三组抗凝剂组骨髓标本各20份,带有PreCP荧光的CD45抗体标记后,用solarbio红细胞裂解液处理后分析残留红细胞与有核细胞总数比例。结果枸橼酸钠、EDTA、肝素钠三组抗凝剂样本三组均值分别为1.70%±1.04%、0.08%±0.14%、1.00%±0.78%。三组间方差分析差异有统计学意义(F值=22.83,P=0.00)。组间差异性分析可知,三组间两两比较,差异均有统计学意义。结论 EDTA抗凝剂对骨髓样本的红细胞裂解效率最好,残留红细胞最低。在进行流式细胞分析时,EDTA抗凝剂作为首选。

Objective To preliminarily discuss the influence of clinical frequently - used anticoagulants （ Heparin sodium, Eth- ylene diamine tetra acetic acid salt and oxalate） on cracking myeloid erythrocytes in the process of cell acquisition and analysis by flow cytometry. Methods Twenty bone marrow specimens were collected from three anticoagulants groups respectively, la- beled with CD45 antibody carrying PreCP fluorescence, treated with Solarbio erythroeyte lysis buffer and then analyzed the ratio of residual erythroeyte and total nucleated cell of the three groups. Results The mean values of oxalate, ethylene diamine tetra acetic acid salt and heparin sodium in three groups were 1.70% ±1.04%, 0.08% ±0.14%, 1.00% ±0.78% respectively. The variance among the three anticoagulant groups had statistical significance（ F =22.83, P =0.00）. Intergroup analysis of the three groups had statistical significance between every two groups. Conclusion EDTA has the highest efficiency on erythroeyte cracking with lowest erythrocyte residues. So EDTA is the most appropriate anticoagulant when doing flow cytometry analysis.