Dojindo,Cellstain- Double Staining Kit/1/CS01,Cellstain-双重染色试剂盒

2022年4月30日 作者 jinpanbio
Cellstain-Double Staining Kit 用于同时对活细胞和死细胞进行荧光染色。该试剂盒包含 Calcein-AM 和碘化丙啶 (PI) 溶液,分别用于染色活细胞和死细胞(图 1)。 Calcein-AM 是一种钙黄绿素的乙酰氧基甲酯,具有高度亲脂性和细胞膜渗透性。虽然 Calcein-AM 本身不是荧光分子,但 Calcein-AM 在活细胞中通过酯酶产生的 calcein 会发出强烈的绿色荧光(激发:490 nm,发射:515 nm)。因此,Calcein-AM 仅对活细胞染色。另一方面,细胞核染色染料 PI 不能通过活细胞膜。它通过死细胞膜的无序区域到达细胞核,并与细胞的DNA双螺旋插入并发出红色荧光(激发:535 nm,发射:617 nm)。由于 calcein 和 PI-DNA 都可以在 490 nm 下激发,因此可以使用荧光显微镜同时监测活细胞和死细胞。在 545 nm 激发下,只能观察到死细胞(图 2)。由于最佳染色条件因细胞系而异,我们建议单独确定合适的 PI 和 Calcein-AM 浓度。请注意,PI被怀疑具有高度致癌性;需要小心处理。

Optimization ProcedureThe following steps may be necessary to determine the suitable concentration of each reagent:

1. Prepare dead cells by 10 minutes incubation in 0.1% saponin or 0.1-0.5% digitonin or by 30 minutes incubation in 70% ethanol.2. Stain dead cells with 0.1-10 μM PI solution to find a PI concentration that stains the nucleus only, not the cytosol.3. Stain dead cells with 0.1-10 μM Calcein-AM solution to find a Calcein-AM concentration that does not stain the cytosol. Then stain viable cells with that Calcein-AM solution to check whether the viable cell can be stained.

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