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To manually enter a gate, display the
graph on which you wish to enter a gate. (For instance, to manually
type in a lymphocyte gate, you might display Forward vs. Side scatter).
If you display a histogram or CDF, then you will be entering a one-dimensional
gate. From the "Graph" menu, select
"Manually enter gate". You will be presented with the
following dialog:
Note that you can enter the gate in units of channels,
absolute intensity, or percentile. FlowJo tells you the minimum
and maximum acceptable values for the gate.
When you select "percentile", then FlowJo
creates a gate based on the percentiles that you enter. For instance,
if you select a gate that has a lower bounds of 50 and upper of
100 (for percentile), then FlowJo constructs a gate which includes
the brightest half of all of the events. Likewise, a percentile
gate of 0 to 5 results in a gate for the dimmest five percent of
the events.
When you make a new gate based on the fluorescence
distribution (percentile), the gate is stored in terms of the channel
numbers. Thus, when you copy the gate to another sample, it is copied
with regard to its position in the window. This way you can make
a gate on the brightest 5% of events, copy it to other samples and
see what fraction of events are above that value in those samples. |
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