multicolor phasor
Hi Munich guys,
We have two-color FRETting single particles and want to be able to correlate the phasor of the Donor channel with e.g. the intensity of the Acceptor channel. We're looking for the best strategy to implement this in Pam.
Pam:
- currently, phasors are calculated per detector, not per PIE channel. This has been annoying me for a long time, so the first suggestion would be to change the phasor part of Pam such that it uses PIE channels rather than detectors.
- Changing the range, and moving the decays with respect to each other would be done similarly as in TauFit. Is this ok for everyone?
- Finally, I would put in an option to simultaneously calculate the phasor for more than 1 channel. Maybe a right-click option in the dropdownbox (is that even possible) to select/deselect that channel from phasor calculation? Otherwise, I just put three dropdownboxes, so user can do phasor up to three channels? Which option do you like most?
ParticleDetection:
- As there's more photons in two PIE channels than for 1, and as particles will overlap 100% on a CLSM, I think it makes sense to do particle detection on the sum of all present channels (or of any defined number of channels in the .phr file). So this option needs to go into ParticleDetection.
- Finally, ParticleDetection or whichever GUI is used now to plot particle intensity vs. lifetime needs to be expanded to be able to plot intensities or phasors from different PIE channels with respect to each other.
Any ideas or suggestions, or extra information on what you are doing in this respect are appreciated!
Anders, why can I not assign Philipp to this, you said Chen and Philipp were both working on Particle detection and phasor?
Greetings, Jelle