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FlowJo allows you to add new parameters to your
data. These parameters are not actually added to the original data
files; they are simply descriptions of new parameters that are stored
with the workspace. However, they behave in other regards like parameters
that were collected: you can display them, calculated statistics
on them, etc.
Because they are not stored with the data file,
they will not appear if you open the same sample in a different
workspace--you will have to add the derived parameters again in
that workspace. Currently, there is no way to copy derived parameter
definitions between workspaces. There are special derived
parameter menu
items.
There are four derived parameter types that you
can create: time, ratio, log->linear, and linear->log.
(Calibrated parameters are also derived parameters, but they are
created using the Calibration
Platform). To add a derived parameter,
select the sample in the workspace, and then select the "Define
new..." menu item attached to the "Derived Parameters..." menu item,
under the workspace menu. You will be presented with a dialog
window that allows you to specify the
parameters to define. From this dialog window, you can also remove
existing derived parameters.
Adding a time parameter is useful for performing
kinetics
analyses. This is usually done when you
have a response that you wish to measure (for example, calcium flux
experiments); however, you might want to see how stable a sample
was during collection, and simply display forward or side scatter
(or fluroescence!) vs. time during collection. Time can only be
added if there is not already a time parameter with the sample.
When you add a time parameter, FlowJo assumes that the event rate
was constant throughout collection, and distributes events evenly
over time. If the start time and end time for acquisition were recorded
by the program collecting the data, then FlowJo knows the time over
which to distribute the events. Otherwise, FlowJo will ask you to
enter how many seconds it took to collect the sample.
You can add a ratio parameter: this is the ratio of any two collected
parameters. This is commonly necessary for calcium flux experiments,
in which the ratio of Indo-1 fluorescence in two different channels
is related to the calcium concentration. You can choose to create
a ratio parameter with either linear or logarithmic scaling.
You can create a calibrated parameter. Using the
"Calibration"
platform you can convert the scaling
of any parameter (for instance, to absolute number of molecules
per cell). While the calibration platform creates a derived parameter
automatically, you may wish to apply such a calibration to other
samples--do this by selecting the appropriate derived parameter.
You can directly select a calibrated parameter (once the calibration
standard exists) by selecting it under the derived parameter submenu.
Finally, you can convert any collected parameter between log and
linear. FlowJo will automatically optimize the scaling of the converted
parameter to show the maximum number of events.
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