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莫名其妙的荧光阳性信号,也可能是荧光素非特异性结合

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发表于 2018-6-19 11:15:48 | 显示全部楼层 |阅读模式

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通常,我们谈及染色的非特异性阳性信号时,通过就是两个原因:死细胞、抗体与细胞上Fc受体非特异性结合,对应的解决方法就是:用死活鉴别染料(如低浓度PI染液或商品化死活鉴别染料)、用Fc Block阻断剂或BSA等。

然而,有时候FLOWER们会发现,即使我老老实实用了上面两个绝招,并且荧光补偿也调节得非常完美了,却仍然还是有莫名其妙的荧光阳性信号,怎么回事?

此时,你可能就得仔细看一下方案,是不是荧光素选错了,导致荧光素非特异性结合,这个问题可能大家都没去关注。
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看了这么多,大家应该知道怎么做了吧,根据自己的实验目的,设计好方案的配色才是第一步,死细胞的排除和Fc阻断,仍应成为常规。



参考文献:
An amazing source for odd questions on flow cytometry is the ‘Cytometry mailing list’ hosted by the Purdue University, which can be found under: https://lists.purdue.edu/mailman/listinfo/cytometry
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