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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 .
OBJECTIVE: To establish a new method for the indirect fluorescent labeling of FCM in peripheral blood.METHODS Anticoagulant whole blood (100 μL) was added to 50 μL of primary antibody and incubated at room temperature for 30 min.The Q- PREP immunological sample preparation instrument, dissolved erythrocytes and fixed standard, add fluorescent secondary antibody 50 μL, reaction at room temperature for 30 min, then add 500 μL of PBS to make a single cell suspension, the flow cytometry analysis, And compared with the other five conventional methods.Results The FCM results of this method and the other five methods were basically the same (P> 0.05), but the method of the present invention has the advantages of less dosage, simple and fast operation, only the dyeing time 70 m in. Conclusions Indirect fluorescent staining of peripheral blood samples (Method 2), which is established by us, is an ideal method for staining FCM samples of peripheral blood. This method provides technical support for the application of FCM in clinical examination and basic research, The practical value.