Understanding the inherent heterogeneity within living systems demands the development of new in vivo single molecule (SM) optical methods to follow protein dynamics without the veil of ensemble averaging. Noble metal nanoclusters exhibit exceedingly strong, size dependent emission throughout the visible and near IR spectrum, but at much smaller sizes (
In Aim II we will synthesize and attach membrane transport functionalities to the nanodots and characterize their uptake and optical properties. These studies lead to Aim III in which multifunctionalized nanodots are made to specifically bind fusion proteins within the cytosol and we gate their transport into specific organelles. The single molecule imaging methods will be developed such that these extremely bright probes can be directly imaged by temporally and spectrally rejecting essentially all background from the more long-lived (ns) autofluorescent species. These combined methods should be capable of increasing current signal/noise ratios by more than three orders of magnitude over current nanoparticle or organic fluorophore based methods. This toolbox of modular, ultrabright, ultrasmall, and short-radiative lifetime nanodots will be generally applicable to a wide range of systems and will be made available to the community through this Exploratory Center.
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