Simultaneous measurement of reactions in microdroplets filled by concentration gradients.
This work describes a technology for performing and monitoring simultaneously several reactions confined in strings of microdroplets having identical volumes but different composition, and travelling with the same speed in parallel channels of a microfluidic device. This technology, called parallel microdroplets technology (PmicroD), uses an inverted optical microscope and a charge-coupled device (CCD) camera to collect images and analyze them so as to report on the reactions occurring in these microdroplets. A concentration gradient of one reactant is created in the microfluidic device. In each channel, a different concentration of this reactant is mixed with a fixed amount of a second reactant. Using planar flow-focusing methodology, these mixtures are confined in microdroplets of pL size which travel in oil as continuous medium, avoiding laminar dispersion. By analyzing the images of parallel strings of microdroplets, the time courses of several reactions with different reagent compositions are investigated simultaneously. In order to design the microfluidic device that consists in a complex network of channels having well-defined geometries and restricted positions, the theoretical concept of equivalent channels (i.e. channels having identical hydraulic resistance) is exploited and developed. As a demonstration of the PmicroD technology, an enzyme activity assay was carried out and the steady-state kinetic constants were determined.
Damean
Nicolae
N
Department of Chemistry, University of Cambridge, Cambridge, CB2 1EW, UK.
Olguin
Luis F
LF
Hollfelder
Florian
F
Abell
Chris
C
Huck
Wilhelm T S
WT
D000469:Alkaline Phosphatase / Q000378:metabolism; D004867:Equipment Design; D004926:Escherichia coli / Q000201:enzymology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011485:Protein Binding; D011489:Protein Denaturation; D013997:Time Factors; D017186:Titrimetry
D000469:Alkaline Phosphatase