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- W2003742864 abstract "FRET-based genetically encoded metal-ion sensors coupled with fluorescence microscopy provide an important tool for real-time imaging of metal-ion homeostasis. The heterogeneity and in vivo kinetics are important sensor properties, yet are less explored because of lacking available experimental methods. We present a high-throughput microfluidic method capable of characterizing the FRET kinetics of metal-ion sensors in mammalian cells, at a throughput of 15 cells/sec with a time window of a few ms to several seconds. Using this method, we screened the HeLa cell Zn2+ sensor library and measured the FRET distribution at single cell level. We also report the quantitative high-throughput cellular-level heterogeneity measurement of cell populations, the cluster map of the Zn2+ sensor FRET change resolves 4 subpopulations with different metal response. Finally, we measured the in vivo sensor kinetics induced by changes in [Zn2+] and [Ca2+]. We observed a > 30-fold rate difference between the extracellular and intracellular sensors, and the kinetics are ionophore-dependent." @default.
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- W2003742864 date "2012-01-01" @default.
- W2003742864 modified "2023-10-18" @default.
- W2003742864 title "Heterogeneity and Kinetics of FRET-Based Ca2+ and Zn2+ Sensors in HeLa Cells" @default.
- W2003742864 doi "https://doi.org/10.1016/j.bpj.2011.11.1139" @default.
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