FCS Express provides the user with many mathematical functions to create custom tokens and custom data sets that can be plotted in Scatter with Regression and Scatter Plots to provide live updating regression curves and fit statistics as gates or data files change.

In this tutorial, we will complete a Spreadsheet, insert a scatter with regression plot, and derive model fit statistics from the data. You will also learn how to create a regression overlay on a scatter plot, how to apply a regression fit to an FCS file, as well as how to save a regression fit for later use.

As an example, we will be performing our analysis on a FITC MESF bead data set in order to obtain an equation that returns the FITC MESF value for a data channel value. Because FCS Express also provides the user with an easy to use Channel Calibration Tool that uses MESF data from a set of beads to calibrate the axes to MESF values, this tutorial is most useful in demonstrating the ways in which custom tokens and custom data sets can be created and plotted and how you can use regression analysis to fit your data in real time right within FCS Express.

In the course of this example, we will:

•Load a layout of MESF beads with histogram markers to complete a Spreadsheet for regression fitting.

•Insert a Scatter with Regression Plot of the Spreadsheet values and automatically generated best-fit regression curve.

•Add a regression fit token to the Scatter with Regression Plot.

•Apply the linear regression to an .fcs data file to obtain MESF data.

•Save the linear regression formula in a text file usable by other .fcs data files and layouts.

 

 

First, we will complete the Spreadsheet and insert a scatter with regression plot.