Development of an immunologically tolerated combination of fluorescent proteins for in vivo two-photon imaging

Combinations of fluorescent proteins (FPs) are routinely used for multi-parameter in vivoimaging experiments to visualize tagged proteins or cell populations of interest. Studies involving FPs are often limited by spectral overlap, toxicity, relative quantum efficiency, and the potential for immunological rejection upon transfer into a non-tolerant recipient. Here we evaluate the immunologic visibility of several commonly used FPs by the murine immune system and identify a spectrally compatible, immunologically tolerated combination of FPs well suited for in vivotwo-photon imaging.