Hello - has anyone used a the cyan-excitable orange emission fluor called CyOFP1
While I understand the reason it may help for 2photon imaging, I’m hesitant about FRET. One of my users just asked for help and he wants to do FRET with mTurq2 and this CyOFP1. I am concerned about correctly separating the donor and acceptor. The fluors are tagged on a protein that they are trying to measure oligimerazation, so some subunits have mTurq2 and some have CyOFP1
CyOFP1 was engineered for multi-color imaging with single excitation (like in 2p FLIM), but I’m not sure how would work for “traditional” acceptor photobleaching or sensitized emission on a laser scanning confocal. Seems like the excitation wavelengths in this proposed case would be too much overlapping to separate donor signal from acceptor signal.
Thanks for any feedback,
Duke Light Microscopy Core Facility