Tomographic imaging of Fluorescence Resonance Energy Transfer in highly light scattering media

摘要

Three-dimensional localization of protein conformation changes in turbid media using Foerster Resonance Energy Transfer was investigated by tomographic fluorescence lifetime imaging. We show that Foerster Resonance Energy Transfer can be localized by reconstructing the quantum yield and lifetime distribution from time-resolved non-invasive boundary measurements. The approach is demonstrated by imaging a highly scattering cylindrical phantom concealing two thin wells containing cytosol preparations of cells expressing a cytosolic genetically-encoded calcium Resonance Energy Transfer sensor. One of the wells contains calcium chloride solution inducing a protein conformation change accompanied with FOrster Resonance Energy Transfer. The resulting fluorescence lifetime distribution, the quantum efficiency, absorption and scattering coefficients were reconstructed.