Fiche publication
Date publication
mai 2010
Auteurs
Membres identifiés du Cancéropôle Est :
Dr MORAS Dino
,
Dr BILLAS Isabelle
Tous les auteurs :
Baret JC, Beck Y, Billas-Massobrio I, Moras D, Griffiths AD
Lien Pubmed
Résumé
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 approximately 7500 cells per concentration provides statistically relevant data that allow precise measurement of the EC(50) (70 nM +/- 12%, alpha = 0.05), in agreement with standard methods as well as with literature data.
Référence
Chem Biol. 2010 May 28;17(5):528-36.
