Reference Standard Calibration
If you choose to use a reference standard to create a Calibrated Parameter, FlowJo shows a window similar to that below. Your graph will be a histogram of the parameter you are calibrating, gated according to how you have created the subpopulation.
When you choose to use the original dynamic range, then FlowJo assumes the calibrated parameter will have the same number of decades of fluorescence as the original parameter (in this example, 4 decades); it only fits a scalar that is used as a multiplier. If you enter absolute units, FlowJo uses the data you enter to determine what the actual dynamic range of the calibrated parameter will be. This will be shown on the left axis. As you enter values, FlowJo draws a line as the regression that lets you see how the original parameter will be converted into the calibrated parameter. In this case, FlowJo will calculate and display the correlation coefficient of the fit.
Once you are satisfied with the fit, click on Define Parameter.Go to the overview of Calibrated Parameters.
