MycoLight 荧光活/死细菌成像试剂盒
| Ex (nm) | - | Em (nm) | - |
| 分子量 | - | 溶剂 | - |
| 存储条件 | - |
MycoLight 荧光活/死细菌成像试剂盒是美国AAT Bioquest生产的用于检测细菌的试剂盒，AAT Bioquest的MycoLight 荧光活/死细菌成像试剂盒提供双色荧光检测，通过荧光显微镜观察活细菌和死细菌。MycoLight 520是一种非荧光酯酶底物，可扩散到革兰氏阳性菌和革兰氏阴性菌中。在细菌细胞内非特异性酯酶水解后，产生绿色荧光产物并在细菌内积累。相比之下，碘化丙锭是一种红色荧光核酸染色剂，只能穿透受损膜的细菌。因此，使用MycoLight 520和碘化丙锭染料的适当混合物，具有完整细胞膜的活细菌发出绿色荧光，而具有受损膜的死细菌产生红色荧光。MycoLight 荧光活/死细菌成像试剂盒是一种用于对活/死细菌进行成像的强大工具。可以在Ex / Em = 488 / 525nm（FITC滤光器组）和540 / 620nm（TRITC滤光器组）下荧光测量染色细胞，分别用于活细菌和死细菌。百萤生物是AAT Bioquest的中国代理商，为您提供优质的MycoLight 荧光活/死细菌成像试剂盒。
适用仪器
| 荧光显微镜 | |
| Ex: | 488/540 nm |
| Em： | 530/620 nm |
| 推荐孔板: | 黑色透明底板 |
| 通道: | FITC/TRITC 通道 |
染色样品示例
概述
1.准备100X MycoLight 520原液
2.准备细菌样本
3.加入MycoLight 520和碘化丙锭
4.将细菌样品与MycoLight 520和碘化丙啶在37°C孵育5-10分钟或在室温下孵育60分钟
5.用FITC和TRITC滤光片组通过荧光显微镜分析样品
注意：在开始实验之前，在室温下解冻所有试剂盒组分
操作步骤
1.MycoLight 520原液（100X）：
将100μLDMSO（组分D）加入MycoLight 520（组分A）的小瓶中。
注意：将储备溶液储存在-20°C，避免光照，反复冻融。
2.制备浓度为106至108个细胞/ mL的细菌样品。 在适当的培养基中将细菌培养到对数晚期。
注意：测量波长= 600 nm（OD600）的细菌培养物的光密度以确定细胞数。 对于大肠杆菌培养物，OD 600 = 1.0等于8×108个细胞/ ml。
4.通过用70％乙醇处理30分钟，然后煮沸60分钟，可以制备死细菌。
5.2充分混合并在37℃下在黑暗中孵育5-10分钟或在室温下孵育60分钟以获得佳染色结果。
5.3通过FITC（Ex / Em = 488 / 530nm）和TRITC（Ex / Em = 540 / 620nm）通道用荧光显微镜监测细菌的荧光。
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