小鼠免疫细胞流式染色过程中细胞丢失量的影响因素  

Factors affecting cell loss in flow cytometry of mouse immune cells

作　　者：郜珊珊[1] 伍俊[2] 周娟[1] 袁祖贻[1] 王丽君[1] GAO Shan-shan;WU Jun;ZHOU Juan;YUAN Zu-yi;WANG Li-jun(Key Laboratory of Environmentand Genes Related to Diseases,Ministry of Education,Xi'an 710061;Department of Neurology,Xi'an Gaoxin Hospital,Xi'an 716000,China)

机构地区：[1]西安交通大学第一附属医院心内科陕西省分子心脏病学重点实验室环境与疾病相关教育部重点实验室,陕西西安710061 [2]西安高新医院神经内科,陕西西安716000

出　　处：《西安交通大学学报（医学版）》2019年第1期134-137,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金青年基金资助项目(No.81500389;No.81700447)~~

摘　　要：目的观察和探讨小鼠免疫细胞多色流式染色过程中细胞丢失的影响因素。方法取C57BL/6J鼠制备脾细胞悬液,分别用不同种类、不同成分和不同pH值缓冲液洗涤和重悬细胞2次,并计数活细胞数量,观察对染色过程中的细胞丢失量有明显影响的因素。结果在小鼠免疫细胞的流式染色标记过程中,缓冲液种类对细胞丢失量有显著影响,含有钾离子的PBS缓冲液洗涤后会造成明显的细胞丢失;离心机转速过高或过低均会导致显著的细胞丢失;而缓冲液pH值的微量偏移和是否加入FBS/BSA对细胞的丢失影响不明显。结论小鼠免疫细胞的流式染色过程中使用含有钾离子的PBS缓冲液会造成明显的细胞丢失,应推荐使用不含钾离子的PBS缓冲液,同时保持离心转速适中。Objective To explore the factors affecting cell loss in multicolor flow cytometry of mouse immune cells.Methods C57BL/6J mice were used in the experiment.Splenocytes were isolated and resuspended twice with buffers of different types,compositions and pH;then the number of viable cells was counted to observe which factors affected the amount of cell loss during staining.Results During flow cytometric labeling of mouse immune cells,buffer types significantly affected the amount of cell loss.Washing with PBS buffer containing potassium ions caused obvious cell loss,while slight shift in buffer pH and whether or not FBS/BSA was added had no significant effect on cell loss.Moreover,centrifugation speed also significantly affected cell loss.Conclusion The use of potassium-containing PBS buffer during flow cytometric staining of mouse immune cells resulted in significant cell loss.The potassium-free PBS buffer and moderate centrifugation speed should be recommended.

关 键 词：小鼠免疫细胞 多色流式标记 细胞丢失 缓冲液成分 

分 类 号：R450[医药卫生—治疗学]