无论您使用哪种染料,四甲基罗丹明甲酯(TMRM)还是MitoTracker Red FM,未处理的细胞都会发出荧光。只要细胞线粒体膜电位降低就会导致荧光信号降低。最重要的是变化的程度。JC-1染料不仅强度改变,还有激发和发射比例光谱的改变。设置未处理的对照和用线粒体膜电位去稳定剂(如CCCP或FCCP)处理的阳性对照是非常重要的。这些染料仅用于活细胞,在固定处理的细胞中无法保留相同程度的信号。
I am seeing high background outside of my neuronal cells when using membrane potential indicators. What can I do to reduce background?
If you use our FluoVolt Membrane Potential Kit (Cat. No. F10488), the kit provides a background suppressor to reduce this problem. For other indicators, consider the use of BackDrop Background Suppressor (Cat no. R37603, B10511, and B10512).