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FlowJo identifies populations (samples,
or gated subsets of samples) by a name that you assign. Naming populations
is very important for the way in which FlowJo works... especially
for batch operations.
The rules for naming gates are similar to what we expect for rules
for naming people in families: siblings cannot have the same name;
however, different generations can have gates with the same name.
When you specify a kind of analysis (for instance,
a statistical measurement, or a gate selecting a subset of cells),
it is applied to a population. During batch operations, you can
specify that this analysis be applied to all samples within a group.
FlowJo decides how to do this by looking at the names of the gates.
For instance, you may have 3 samples in a workspace.
You first decide on a gate to exclude dead cells and clumps of cells;
you apply this gate to a group containing all three samples (and
call it "Live cells"). After looking at the 3 samples,
you find that you have to alter the position of the gate slightly
for each one--therefore, the gates are no longer identical.
Now you create a statistic that tells you the median
forward scatter signal for "Live cells." Thus, you create
a new node (analysis) attached to the "Live cells" gate
of the first sample. When you now drop this node on the group's
version of "Live cells", the node is attached to each
sample's version of its own "Live cells" gate. Even though
each gate is slightly different, FlowJo assumes that the gate represents
the same population and thus you want the same operations applied
to that gate.
This is why a gate cannot have the same name as
another gate of the same population: FlowJo would not be able to
decide which gate you want analyses applied to. Note that FlowJo
decides which gate you mean by not only name, but generation. Thus,
if you have a sample which has a "Live Cells" gate as
a child of a gate you named "Lymphocytes", this is NOT
the same gate as "Live Cells" attached directly to the
sample--and, in the example above, would not have your statistical
analysis applied.
Realize that the complete name of any population
is actually the concatenation of all of its ancestors' names with
its own name. Thus, in a sample, if you have created a lymphocyte
gate, and within that a CD3 gate, then that subpopulations "true"
name is "Lymphocyte/CD3". FlowJo only shows you the "CD3"
portion of the name, as the "Lymphocyte" portion is implied
by the indentation in the workspace. You could subset the "CD3"
population and also call it CD3. This is legal, because its full
name would be "Lymphocyte/CD3/CD3" which is distinct from
"Lymphocyte/CD3". However, it could be confusing--so try
to avoid duplicate gate names at any level.
If you wish, you may get help on how to create new
subpopulations by gating.
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