Lower convolution threshold - how low or high it can be
Hello Cytofers,
I wanted to start subject about lower convolution threshold. I am working on Cytof2 upgraded to Helios. I know that a default setting is 400, however it will depends also on the amount of channels that we will open in the particular experiment, right?
My question is how you are dealing with this?
I have one experiment when I am measuring cytokines response, so my panel for now is about 20 markers, however I would like to apply something like MMM (mass minus multiple) control in order to see how to put the gates, so in the panel of MMM I would have 9 AB less. I assume that I would need to change the lower convolution threshold, as I will have less channels open?
And is playing with the threshold really helps with getting rid of the debris and instead acquiring more data?
I have also heard that sometimes people are using threshold around 1000 which compare to the one suggested by fluidigm sounds high…. I have acquire few samples with around 20 markers (including cytokines) at 1000 and now I am wondering if that was OK and if by using IMD files I can still re-analyze my data at let’s say threshold 400. And if I do so, how exactly I can verify if one or the other threshold works better ?
Best,
Paula
I wanted to start subject about lower convolution threshold. I am working on Cytof2 upgraded to Helios. I know that a default setting is 400, however it will depends also on the amount of channels that we will open in the particular experiment, right?
My question is how you are dealing with this?
I have one experiment when I am measuring cytokines response, so my panel for now is about 20 markers, however I would like to apply something like MMM (mass minus multiple) control in order to see how to put the gates, so in the panel of MMM I would have 9 AB less. I assume that I would need to change the lower convolution threshold, as I will have less channels open?
And is playing with the threshold really helps with getting rid of the debris and instead acquiring more data?
I have also heard that sometimes people are using threshold around 1000 which compare to the one suggested by fluidigm sounds high…. I have acquire few samples with around 20 markers (including cytokines) at 1000 and now I am wondering if that was OK and if by using IMD files I can still re-analyze my data at let’s say threshold 400. And if I do so, how exactly I can verify if one or the other threshold works better ?
Best,
Paula