It is often useful to look at many samples on the same plot to see sample variation and to compare experimental samples against a control. In this section, you will learn how to overlay multiple kinetics samples, apply parameter math to the overlays, and format the overlays to make the data easier to view.

Although any kinetics data files of your own can be used to follow along, you have the option of using the same sample data files used in this tutorial. The sample data files are located in the Tutorial Sample Data archive. The KineticsTutorialFormatting.fey, kinetics8.008, kinetics9.009, kinetics10.010, and kinetics25.025 data files will be used here.

 

1. Select File tab→Open Layout.

2. Select the KineticsTutorialFormatting.fey layout file located in the FCS Express Sample Data directory from the Load Layout dialog.

 

The layout contains one kinetics plot displaying Time vs Fluo 3 for sample kinetics25.025. We will now add the other three samples to the kinetics plot as overlays.

3. Ctrl+Click on samples kinetics8.008, kinetics9.009, and kinetics10.010 to select them in the data list.

4. Drag the files from the data list onto the Kinetics Overlay plot.

5. Release the mouse button to drop them on the plot.

6.Choose add the files to the plot as new overlays.

7.Click OK.

 

The four sample are now displayed simultaneously in the kinetics plot. Though, the Parameter Math transformation is not yet applied to the plot. We will now apply the Parameter Math transformation to the plot via drag and drop.

8.Left click and hold on the New Parameter Math transformation from the Transformation window.

9.Drag the New Parameter Math transformation out of the Transformation window and release it on top of  the kinetics plot (Figure T21.5). The Parameter Math applied suffix will be added to the plot title as a confirmation of this action.

10. Change he Y-Axis parameter on the Kinetics Overlay plot to Ca++ Ratio.

11. Select the Live Cells gate from Gating→Create Gates→Current Gate section of the Ribbon bar.

The plot now contains overlays for each sample and should look like Figure T21.19.

 

 

We will now format the plot to make it more visually appealing.

18. Right-click on the plot.

19. Choose Format from the pop-up menu.

20. Choose the Overlays category.

21. Ctrl+Click on all of the plots in the list box to select them.

22. Change Line Options→Width to 1.50.

23. Choose the Legend category.

24. Click on the Visible check box to select it.

25. Change the Position from Right to Bottom.

26. Click OK.

The plot should now look like the Figure T21.20.

 

 

Finally, we will change the resolution of a kinetics plot.