Calibrate Alignment of Green / Red Channel

[BradyAJohnston]

I'm wondering if there is a way to calibrate the left / right channels - given that during the experiment they were not perfectly symmetrical. In other software that we were previously using for this, we could calibrate with a standard - but it seems that deepFRET just divides the sensor in half and overlays them without any possibility for adjustments?

Example image below, with highlighted spots that are the more obvious examples of spots that should align circled in pink.

Because of this off-alignment, the spots that do 'co-localise' are just jibberish as the signals belong to different molecules.

Any chance that this is possible with deepFRET? The raw data is just .tif movies that look like below.

[graciesiffer]

We are having the exact same problem, did you ever find a solution?

[BradyAJohnston]

@graciesiffer sadly not! I ended up abandoning this approach as it was unusable for this type of data in the current state.