流式细胞术测定红细胞存活率的方法建立

Establishment of Method to Determine Survival Rate of Red Blood Cells by Flow Cytometry

目的 :建立应用流式细胞术测定红细胞体内存活率的方法。方法 :献血者Rh表型ccDEE ,受血者为CCDEE ,以抗 c为一抗结合ccDEE的红细胞 ,两群细胞按设定的比例混合 ,以山羊抗人标记荧光素的IgGab段为二抗连接抗 c ,经流式细胞仪测定ccDEE表现型红细胞在两群细胞中所占的比例 ,并与实际的比例进行相关性比较。结果 :已知两群细胞数的比例与流式细胞仪测定的比例的相关系数r =99.87。结论

Objective:To establish the method to determine the in vivo survival rate of red blood cells(RBCs)by flow cytometry.Methods:The Rh phenotype of donor′s RBCs was ccDEE,and that of recipient′s RBCs was CCDEE.Anti c antibody was taken as the first antibody binding the donor′s RBCs.Then,two groups of RBCs were mixed in accordance with the given ratio.The goat anti human IgG ab fragment labelled by fluorescein was taken as the second antibody linking the anti c.The ratio of ccDEE RBCs in two groups of RBCs was determined by the flow cytometry and was correlatively compared with the actual ratio.Results:It was known that the correlation coefficient between the actual ratio of RBCs counts in two groups and the ratio of those determined by flow cytometry was r =99.87.Conclusion:The flow cytometry could be used to perform the in vitro determination of in vivo survival rate of RBCs.