Example with 5-Color Panel

Here's a quick introduction with a set of demo files that will get you up and running.

1. Start the V-Comp™ app

If it is the first time you have run the software, you will be asked to register it. Click here to see how.

2. Create a new V-Comp™ setup

If you have never used the software before, the V-Comp™ Setup Wizard will launch automatically. If you don't see the wizard dialog, click the New button to create a new setup.
On the Welcome page, select Basic for the setup type.

A Basic setup uses one single-stained control file for each of the fluorescence markers in the panel, which is the case for this demo. A Label-specific setup can have more than one single-stained file for the fluorescence markers, and a Spectral setup is used for unmixing data collected on a spectral cytometer.
After selecting the setup type, click to move to the next pane: Single Color Controls.
Type a name for your new setup. For this example, type "Demo Setup".
Next we need to select the single-color control files. For this example, click the button and navigate to the location on your computer where the V-Comp™ app is installed. On the Mac, that's usually in the "Applications/V-Comp for Mac/Samples" folder. On the PC, look in "Program Files (x86)\Verity Software House\V-Comp for Window\Samples".
Select the 5 single-color controls in the Samples folder as shown below. For this example, these are fluorescent protein controls.
Click to add the single-color control files. The order of the files does not matter. You can also drag-and-drop files onto the Single-Color Controls list to add them.
Click to move to the Unstained Controls pane.
On this pane, you can drag and drop your unstained control file onto the wizard or click the folder icon to select one. You can set a different unstained for each file, but that's not usually needed.
Click and select the file named "Unstained Control.fcs".
Click to add the file. Notice that the file is associated with all of the single-color controls. In working with your own files, you can specify a separate unstained for any of the single-color files. For this example, we only need the one unstained file.
Click to move to the Detectors pane.
The program will try to figure out which detectors you want to use for compensation automatically.
In our demo, the correct detectors will be selected, but always review the selections and make sure the correct detectors are selected. These are the parameters that will be compensated.
Click to move to the Match Controls with Detectors pane.
The program will assign a control file to the detector that was brightest for that control. Most of the time, this is correct, and it is correct for our demo files.
If you need to change assignments, drag the single-color controls to the correct detector in the list.
Click to move to the Options pane.
On this page, you can edit the setup name and description and select several other options for the setup.
Set the Export Option to "Export files to the Export Path". This means we want to use a specific location for the files we export.

Note: make sure to select the correct option here for this example. Choose the "Export files to the Export Path" option so that you can select a writable folder on your computer.
Click the icon to the right of Export Path and select a folder for which you have write-permissions.

Note: you must be able to create and write files in the path you select here.
Set the other options as shown.
Click to complete the wizard.
The wizard will analyze your control files and create a V-Comp™ compensation setup. You should see a Congratulations dialog when it finishes.
Click the link to see the report and the program will show a detailed report about the V-Comp™ compensation setup you just created. This report is automatically stored in the Reports folder. Review the report and the on-screen instructions.
Click OK to close the dialog. You have successfully created a V-Comp™ compensation setup from a set of control files.

3. Apply the V-Comp™ setup to your files

When you create a new V-Comp™ setup with the wizard, the new setup is automatically selected and you are ready to apply it to files acquired with the panel.

Click the button and in the Open dialog, navigate to the same folder where the control files for this example were located. Select the "All Color" files in the folder and click Open.
As files are added to the batch list, you should see a symbol next to each one. This indicates that the files are compatible with the setup you have selected. If you see files with a symbol, they are not compatible with the current setup and will not be processed.
Click the button to process the files in the list. The V-Comp™ app will export a new version of each one with the new compensation settings. Your original files are left untouched. A report of the export process is automatically created and stored in the Reports folder.

The files that the V-Comp™ app creates contain a new $SPILLOVER keyword which defines the new compensation. Most software packages can read these files and interpret the new compensation settings, so you can perform your data analysis with your favorite software tools.

Summary

To create a new V-Comp™ compensation setup, all you need is a set of single-color controls and one unstained control file created with the instrument settings for your panel. The unstained control needs to be the same particles (cells, beads, etc) as the single-stained controls. Use the V-Comp™ Setup Wizard to create a new setup. Once a setup is created, it can be reused with any data files that were acquired on the same instrument with the same fluorochromes and the same settings.

To apply V-Comp™ compensation to your all-color FCS files, first select the V-Comp setup, then add your all-color files to the batch list and click the button. New files will be created with the new compensation keyword, ready to be analyzed with your favorite analysis tools.