Live or Dead 细胞活性检测试剂盒是美国AAT Bioquest生产的细胞活性 检测试剂盒，AAT Bioquest的Mycolight 细菌活力测定试剂盒可在革兰氏阳性和阴性细菌中提供细菌活力的双色荧光测定。该试剂盒利用我们的绿色荧光核酸染色剂MycoLight Green和红色荧光核酸染色碘化丙啶的混合物。单独使用时，MycoLight 绿色污渍通常会标记群体中的所有细菌（活的和死的）。相比之下，碘化丙锭只穿透膜受损的细菌，当两种染料都存在时，导致MycoLight 绿色染料荧光的减少。因此，使用MycoLight 绿色和碘化丙啶的适当混合物，具有完整细胞膜的活细菌发出绿色荧光，而具有损伤膜的死亡或濒死的细菌 产生红色荧光。Mycolight 细菌活力测定试剂盒是监测细菌种群活力的强大工具，可作为细胞膜完整性的函数。可以在510-530nm（FITC滤光片）和600-660nm（德克萨斯红色滤光片光片）处以488nm处（最常见的激发光源）激发荧光测定染色的细胞。

样品实验方案

简要概述

1.用测试化合物制备细胞
2.添加染料加工溶液
3.在室温或37°C孵育30分钟至1小时
4.在Ex / Em = 610/650 nm（截止= 630 nm，红色）和Ex / Em = 360/450 nm（截止= 420 nm，蓝色）或在荧光显微镜下观察（Texas red/Cy5通道   活细胞），（DAPI通道  死细胞）

工作溶液配制

将5μL的200X Cellbrite Red（组分A）和5μL的200X Nuclear Blue DCS1（组分C）加入1mL的测定缓冲液（组分B）中并充分混合以制备染料 - 工作溶液。 该染料加工溶液在室温下稳定至少1小时。 注意：由于最佳染色条件可能因细胞类型的不同而异，因此建议单独确定组分A和C的适当浓度。

操作步骤

1.根据标准方案准备细胞。 注意：我们用星形孢菌素（SS）在37℃下处理HeLa细胞4小时以诱导细胞凋亡。 详细信息请参见图1。

2.用100μL/孔（96孔板）或25μL/孔（384孔板）的染料加工溶液替换生长培养基。

3.将染料加工溶液板在室温或37°C孵育30分钟至1小时，避光。

4.用HHBS，PBS或您选择的缓冲液洗涤细胞两次。

5.向细胞中加入100μL/孔（96孔板）或25μL/孔（384孔板）的测定缓冲液（组分B）。

6.用荧光显微镜监测荧光信号，用德克萨斯红或Cy5过滤器检测活细胞，用DAPI过滤死细胞。 还可以使用荧光酶标仪（底部读取模式）在Ex / Em = 610 / 650nm（截止= 630nm，红色）和Ex / Em = 360 / 450nm（截止= 420nm，蓝色）下分析荧光强度。

数据分析

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