Colour Compensation

Colour compensation

Some devices and/or assays require extra calibration. When analyzing multiplex assays (multiple fluorophores per well) on the LightCycler® 480 or Roche Cobas®, it is often necessary to perform a colour compensation experiment. This experiment corrects the fluorescence crosstalk between the channels.

How can I add a colour compensation file?

1. Click + in the device module
2. Select a directory and a CC file

• Only CC files from the selected device can be uploaded
• Upload the CC experiment file itself, not the CC object
• When uploading a .xml file, CC sample names should be the same as the dominant channels (the channel in which fluorescent molecules are present during the colour compensation experiment) i.e. ‘excitationWL-emissionWL’; water sample names should be ‘Blank’ (case-insensitive)

1. Select the CC subset containing the CC sample positions
2. Click NEXT
3. Link your CC file to the correct assay(s) (you can only select assays that are already added to your Assays module) and choose if you want the CC file as default
4. Click SAVE

• The added CC file will now be visible (also in the linked assay information)
• The added CC file will have a green checkmark if it was set as default

How can I edit a colour compensation file?

1. Hoover over the CC file
2. Click the EDIT ICON (pencil)
3. You can now inactivate your CC file, change the linked assay(s) or the default settings
4. Click SAVE

• The added CC file will be grayed out if it is inactivated
• The added CC file will have a green checkmark if it was set as default