التفاصيل البيبلوغرافية
| العنوان: |
High-Throughput Examination of Fluorescence Resonance Energy Transfer-Detected Metal-Ion Response in Mammalian Cells. |
| المؤلفون: |
Hairong Ma1,2, Gibson, Emily A.1, Dittmer, Philip J.1, Jimenez, Ralph1,2 rjimenez@jilau1.colorado.edu, Palmer, Amy E.1 amy.palmer@colorado.edu |
| المصدر: |
Journal of the American Chemical Society. 2/8/2012, Vol. 134 Issue 5, p2488-2491. 4p. |
| مصطلحات موضوعية: |
*BIOSENSORS, *FLOW cytometry, *FLUORESCENCE resonance energy transfer, *CELLS, *METAL ions, *MICROFLUIDICS |
| مستخلص: |
Fluorescence resonance energy transfer (FRET)-based genetically encoded metal-ion sensors are important tools for studying metal-ion dynamics in live cells. We present a time-resolved microfluidic flow cytometer capable of characterizing the FRET-based dynamic response of metal-ion sensors in mammalian cells at a throughput of 15 cells/s with a time window encompassing a few milliseconds to a few seconds after mixing of cells with exogenous ligands. We have used the instrument to examine the cellular heterogeneity of Zn2+ and Ca2+ sensor FRET response amplitudes and demonstrated that the cluster maps of the Zn2+ sensor FRET changes resolve multiple subpopulations. We have also measured the in vivo sensor response kinetics induced by changes in Zn2+ and Ca2+ concentrations. We observed an ∼30 fold difference between the extracellular and intracellular sensors. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
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