I've been confused about how to setting the trigger channel and threshold in the nanocellect program. I have the GFP positive and negative cell which I've confirmed by the confocal, the positive one looks bright green under the confocal. But when I try to sort the cell by using the Wolf nanocellet, it couldn't separate the positive and the negative cells.

So I need to know that, when I first flow the negative sample into the nanocellect what trigger channel should I use? and also the threshold, what number should I set?

And when I flow the GFP positive sample in, should I use the same trigger channel as in the negative one?