Hi there,
Any help on this possibly easily solved query will be gratefully received!
I'm looking to analyse FCS files on FlowJo - the dot plot files display correctly but when I change the axis to histogram, all the data is centered around 0? This is not a problem with the actual FCS files as they will display correctly on other computers running the software. I have tried altering the axis etc but the problem seems to be with the actual data. I have attached a file to show you what I mean.
Many thanks
Philana
First of all I have to mention that I don´t know the software.
But you are displaying forward scatter vs. sideward scatter in the dot plot. The channal for the histogram seems to differ (488 A-2). It might be the fluorescence signal at 488 nm.
Could that be the problem?
Best Björn
Thanks for your answer Bjorn - no I just attached the dot plot to show that the data does appear to be there (for the FSC and SSC) anyway but when I plot as a histogram for my fluorophore of interest, I get the thin line centered around zero.
Could you provide a 488 vs FFC dot plot? This could be helpful to get an impression what is goin wrong.
An additional a FFC histogram could validate if it is a software problem.
These figures clearly explains the previous results.In the second figure, you see that there is no software problem. The histogram well reproduces the dot plot data.
From the first figure, you can see that there is no signal in the 488 chanal. For all events (dots in the 488 vs FFC plot) the 488 value is around zero. If you make a histogram of a lot of zeros, it had to be a spike centered at zero.
Again, this seems not to be a software problem. There is simply no signal in your 488 channel. This should be an experimental / flow cytometer problem.
Was this helpful?
Björn
PS: I assume, that you don't have active gates, that causes the problems?
Hi Bjorn, I appreciate your help. Unfortunately, the data is displaying correctly on other computers so it isn't a problem with the data itself. There is signal in the 488-1 channel but simply is not displaying correctly.
Hi Philana,
I met same issues when open data with flowjo, how do you find out the solution in the end?
Thanks
Pengfei
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