期刊
CHEMISTRY & BIOLOGY
卷 17, 期 5, 页码 528-536出版社
CELL PRESS
DOI: 10.1016/j.chembiol.2010.04.010
关键词
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资金
- European Molecular Biology Organization
- European Commision Framework 6 (EC FP6) MiFem Network
- Ministere de l'Enseignement Superieur et de la Recherche
- Centre National de la Recherche Scientifique (CNRS)
- Agence National de la Recherche (ANR) [ANR-05-BLAN-0397]
- Agence Nationale de la Recherche (ANR) [ANR-05-BLAN-0397] Funding Source: Agence Nationale de la Recherche (ANR)
We used a droplet-based microfluidic system to perform a quantitative cell-based reporter gene assay for a nuclear receptor ligand. Single Bombyx mori cells are compartmentalized in nanoliter droplets which function as microreactors with a >1000-fold smaller volume than a microtiter-plate well, together with eight or ten discrete concentrations of 20-hydroxyecdysone, generated by on-chip dilution over 3 decades and encoded by a fluorescent label. The simultaneous measurement of the expression of green fluorescent protein by the reporter gene and of the fluorescent label allows construction of the dose-response profile of the hormone at the single-cell level. Screening similar to 7500 cells per concentration provides statistically relevant data that allow precise measurement of the EC50 (70 nM +/- 12%, alpha = 0.05), in agreement with standard methods as well as with literature data.
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