Here are instructions showing how to use 405 nm (isosbestic signal) to remove motion artifacts from a red signal in danse:
- Decimate signal (not necessary if using default Lock-In settings in DNS, which automatically decimate by 200x)
- Baseline Correction (remove exponential decay due to photobleaching to flatten the signal)
- Align signals (this standardizes isosbestic and biosensor signal so that they can be properly subtracted)
- Subtract isosbestic from biosensor signal
- Filter the signal to remove high-frequency noise (optional)
See example of this is correction, which successful calculated dFF for red signal and remove of motion artifacts.
See the example of pipeline and parameters used in danse™ software to correct the above example: