This app can quickly give the equation needed for FlowJo’s ‘derive parameter’ to convert fluorescence arbitrary units to MESF units.
Please leave a comment below if there are any issues using the software or if there are any additions that would be useful in future versions.
Steps to use:
1. Open the app and enter the bead MESF units. In FlowJo gate around each of the fluorescent peaks and enter the corresponding Median channel number for each peak into the MESF calculator. Finally, add the name of the parameter that MESF units are being calculated for in FlowJo.
2. Click ‘Calculate’.
3. In FlowJo, click on a sample in the workspace, go to the Instrumentation band under the Workspace tab (if FlowJo is running in Experienced mode) or go to the Instrumentation band under the Tools tab (if FlowJo is running in Advanced mode) and select Derived Parameters. This will bring up the Derived Parameter window (below):
4. Copy the formula to the right of the equals sign from step 2 and paste into the formula box in the window above and click ‘ok’. The scale may need adjusting to ensure all data is visible. The derived parameter now represents the calculated molecules of the equivalent soluble fluorophore.