|
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 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.
Click here to view the special derived parameter menu
items.
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 fluorescence!) 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.
|