流式细胞术外周血样品间标的方法

Indirect staining of peripheral blood samples for flow cytometry

目的 建立一种新的用于流式细胞术 (FCM)外周血样品的间标荧光染色方法 .方法 抗凝全血 1 0 0 μL,加入一抗 5 0μL ,室温下反应 30 min,上 Q- PREP型免疫学样品制备仪 ,溶解红细胞和固定标品 ,加入荧光二抗 5 0 μL ,室温下反应 30 min,再加入 5 0 0 μL 的 PBS制成单细胞悬液 ,上流式细胞仪分析 ,同时与其他 5种常规方法进行比较 .结果 本方法与其他 5种方法的 FCM测定结果基本一致 (P>0 .0 5 ) ,而本方法样品用量少 ,操作简单 ,快速、染色时间仅需 70 m in.结论 我们建立的外周血样品间接荧光染色法 (方法 2 ) ,是外周血 FCM样品染色的理想方法 ,该方法对于 FCM在临床检验和基础研究的应用提供了技术支持 ,具有较强的实用价值 .

AIM To establish a new method for indirect fluorescent staining and labeling of peripheral blood samples for flow cytometry (FCM). METHODS 50 μL of first antibody was added to 100 μL of anticoagulant peripheral blood before it was incubated for 30 min. at room temperature. Then the sample was put on the Q PREP immunological sample preparatory instrument in order to dissolve red cells and fix the sample. 50 μL of second fluorescent antibody was added to the sample. Then the sample was incubated for 30 min. at room temperature before 500 μL of PBS was added to make a unicellular suspension. The suspension underwent flow cytometry (FCM), the result of which was compared with that of five other conventional staining methods. RESULTS No significant difference was found between/among FCM 2 dimensional scatter diagrams and results of the assessment of the positive cells of the samples by the 6 methods. In comparison with the protocols of the other five methods, this method (Method 2) proved to be simple, convenient and economical. CONCLUSION Indirect fluorescent staining of peripheral blood samples is optimal for the assessment of peripheral blood samples by FCM. It can be of great help and use in the application of FCM in clinical examination and basic medical research.