Fig. 1
From: Out-of-equilibrium microcompartments for the bottom-up integration of metabolic functions
Microfluidic platform for monitoring compartmentalized metabolic reactions. a Microfluidic workflow. Biochemical components are encapsulated in 30 pL w/o droplets stabilized by a block-copolymer surfactant. Different droplet compositions are barcoded with a fluorescent dye. Compartmentalized reactions are activated by picoinjecting a metabolic substrate or cofactor. The microcompartments are incubated on-chip to monitor their metabolic state using fluorescent readouts. Scale bars 100 μm. b, c Kinetics of compartmentalized reactions. b G6PDH activity. Glucose-6-phosphate dehydrogenase (G6PDH) oxidizes d-glucose-6-phosphate 1 (G6P) into 6-phospho-d-glucono-1,5-lactone 2 (GLP) with the concomitant reduction of NAD+ into NADH. NADH concentration versus time (t) of 30 pL w/o droplets containing NAD+ (250 μM) and G6PDH 0, 0.01 and 0.08 U mL−1 (green, yellow, and red curves, respectively) or NADH 200 μM internal reference (black) after injection of G6P substrate (1 mM). Error bars are defined as s.d. (N = 2000). c 2D histograms of NADH fluorescence versus barcoding fluorescence (30, 60, 90, and 120 μM sulforhodamine B, respectively) at different incubation times for the 4-bit emulsion. Reactions are performed in NaOH-Tricine buffer (100 mM, pH 8.0) with MgCl2 5 mM
