| 摘要: |
| 建立和优化了流式细胞术检测虾类血细胞活性氧含量的方法。以2′,7′-二氢二氯荧光黄双乙酸钠(DCFH-DA)为ROS 特异探针, 以斑节对虾(Penaeus monodon)血细胞为材料, 用流式细胞仪检测不同染料浓度和不同染料孵育时间下血细胞的荧光强度、细胞形态特征和细胞活性的变化。结果表明, 利用FCM 检测虾类血细胞ROS 含量的适宜的染料浓度和最佳的孵育时间分别为10 μmol/L 和30 min。 |
| 关键词: 流式细胞术 活性氧 虾 血细胞 |
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| 基金项目:国家自然科学基金资助项目(30671628); 广东省教育部产学研结合资助项目(2006D90204005); 国家科技支撑计划项目(2007BAD29B06); 广东省科技计划项目(2007B020708013); 广东省重点科技项目(2009A020101006); 广州市科技支撑计划项目(2009Z1-E681) |
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| Measurement of reactive oxygen species (ROS) production in shrimp haemocyte by flow cytometry |
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| Abstract: |
| In this study, a flow cytometric method used to measure the reactive oxygen species (ROS) production in shrimp haemocytes was improved. 2′,7′-dichlorofluorescin (DCFH-DA) was used as a probe of ROS, and Penaeus monodon haemocyte was used as material. The cellular DCF fluorescent intensity, morphologic parameter and viability incubated with different doses of probe for different time were measured using flow cytometer. The results show that the optimal DCFH-DA concentration and incubation time for shrimp haemocyte ROS measurement using flow cytometry were 10 μmol/L and 30 min, respectively. |
| Key words: flow cytometry reactive oxygen species shrimp haemocyte |
