When I do the gating in the flow cytometer, does it just record the part that is in the gate? could I do the run without gating in the piece of equipment and then do that in the software?
Not sure what instrumentation you are running. But in general, on an BD analysis instrument running the Diva software, you are collecting all the information about any sample in the .FCS file. You can always go back in and re-gate after the fact. However, you may want to adjust the populations you are looking at and change laser settings or compensation, which is harder to do afterwards.
Not sure what instrumentation you are running. But in general, on an BD analysis instrument running the Diva software, you are collecting all the information about any sample in the .FCS file. You can always go back in and re-gate after the fact. However, you may want to adjust the populations you are looking at and change laser settings or compensation, which is harder to do afterwards.
It depends on your cytometer software. In general, drawing the gates on your flow cytometer does not limit the recording to what's in the gate, although there may be a setting to do that. In FACS Diva, there is a setting on the Acquisition Dashboard called "storage gate". Make sure it is set to "all events" if you want to record everything. Otherwise, you can set it to a particular gate if you want a smaller file size.
You can indeed do the run without gating on the cytometer. However, I usually draw some rough gates on the machine because it is nice to be able to get a quick idea of the results while you are running it. Also, you will quickly realize if you ought to adjust the voltage/gain instead of wasting all the samples and only finding out later that e.g. one population is off the screen.
I agree with David, this is also the case on the Partec cytometer we have!! The machine gathers all the information available and you can always modify your gates in a later timepoint.
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