Hello,
I created a region (R0) in a dot plot. Then I gated a second dot plot with the first region R0, and I created a second region called R1. Then I created histogram and I tried to apply the gate called R1, but it shows me the data for the full first dot plot. I'm clearly doing something wrong, but I don't know what.
Any advice?
Best,
JJ
Regions do not build a parent-child hierarchy in FCSalyzer. This id different from other analysis software, and reflects how the old CellQuest software worked.
So even though the plot you used to draw R1 was gated on R0, a gate just using R1 will filter all events, without a pre-filtering by R0.
You need to create a gate with a boolean strategy: "R0 and R1"
That gate will filter the events using both regions.
If you would like to refer to this comment somewhere else in this project, copy and paste the following link:
Anonymous
Anonymous
-
2021-07-13
Hello Sven,
Thank you very much for your reply. It's awesome. I'm going to try it. You are totally right. We are so used to FlowJo, CellQuest, etc, that I assumed that FCSalyzer worked the same way.
And thank you very much for creating this wonderful free software for flow cytometry. We really needed a free alternative.
Best regards,
JJ
If you would like to refer to this comment somewhere else in this project, copy and paste the following link:
Hello,
I created a region (R0) in a dot plot. Then I gated a second dot plot with the first region R0, and I created a second region called R1. Then I created histogram and I tried to apply the gate called R1, but it shows me the data for the full first dot plot. I'm clearly doing something wrong, but I don't know what.
Any advice?
Best,
JJ
Hi JJ,
Regions do not build a parent-child hierarchy in FCSalyzer. This id different from other analysis software, and reflects how the old CellQuest software worked.
So even though the plot you used to draw R1 was gated on R0, a gate just using R1 will filter all events, without a pre-filtering by R0.
You need to create a gate with a boolean strategy: "R0 and R1"
That gate will filter the events using both regions.
Please have a look at the tutorial: https://youtu.be/RWIh2mgQCcM
Regards,
Sven
Hello Sven,
Thank you very much for your reply. It's awesome. I'm going to try it. You are totally right. We are so used to FlowJo, CellQuest, etc, that I assumed that FCSalyzer worked the same way.
And thank you very much for creating this wonderful free software for flow cytometry. We really needed a free alternative.
Best regards,
JJ