We present a single particle fluorescence resonance energy transfer (spFRET) study of freely diffusing self-assembled quantum dot (QD) bioconjugate sensors, composed of CdSe-ZnS core-shell QD donors surrounded by dye-labeled protein acceptors. We first show that there is direct correlation between single particle and ensemble FRET measurements in terms of derived FRET efficiencies and donor-acceptor separation distances. We also find that, in addition to increased sensitivity, spFRET provides information about FRET efficiency distributions which can be used to resolve distinct sensor subpopulations. We use this capacity to gain information about the distribution in the valence of self-assembled QD-protein conjugates and show that this distribution follows Poisson statistics. We then apply spFRET to characterize heterogeneity in single sensor interactions with the substrate/target and show that such heterogeneity varies with the target concentration. The binding constant derived from spFRET is consistent with ensemble measurements.
Time resolution of current single-molecule fluorescence techniques is limited to milliseconds because of dye blinking and bleaching. Here we introduce a photoprotection strategy that affords microsecond resolution by combining efficient triplet quenching by oxygen and Trolox with minimized bleaching via the oxygen radical scavenger cysteamine. Using this approach we resolved the single-molecule microsecond conformational fluctuations of two proteins: the two-state folder α-spectrin SH3 domain and the ultrafast downhill folder BBL.
Abstract. An earth system model has been developed at Beijing Normal University (Beijing Normal University Earth System Model, BNU-ESM); the model is based on several widely evaluated climate model components and is used to study mechanisms of ocean-atmosphere interactions, natural climate variability and carbon-climate feedbacks at interannual to interdecadal time scales. In this paper, the model structure and individual components are described briefly. Further, results for the CMIP5 (Coupled Model Intercomparison Project phase 5) pre-industrial control and historical simulations are presented to demonstrate the model's performance in terms of the mean model state and the internal variability. It is illustrated that BNU-ESM can simulate many observed features of the earth climate system, such as the climatological annual cycle of surface-air temperature and precipitation, annual cycle of tropical Pacific sea surface temperature (SST), the overall patterns and positions of cells in global ocean meridional overturning circulation. For example, the El Niño-Southern Oscillation (ENSO) simulated in BNU-ESM exhibits an irregular oscillation between 2 and 5 years with the seasonal phase locking feature of ENSO. Important biases with regard to observations are presented and discussed, including warm SST discrepancies in the major upwelling regions, an equatorward drift of midlatitude westerly wind bands, and tropical precipitation bias over the ocean that is related to the double Intertropical Convergence Zone (ITCZ).
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