Interfacing Microwells with Nanoliter Compartments: A Sampler Generating High-Resolution Concentration Gradients for Quantitative Biochemical Analyses in Droplets

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Publikace nespadá pod Ekonomicko-správní fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
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GIELEN Fabrice BURYŠKA Tomáš VAN VLIET Liisa BUTZ Maren DAMBORSKÝ Jiří PROKOP Zbyněk HOLLFELDER Florian

Rok publikování 2015
Druh Článek v odborném periodiku
Časopis / Zdroj Analytical Chemistry
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www http://pubs.acs.org/doi/abs/10.1021/ac503336g
Doi http://dx.doi.org/10.1021/ac503336g
Obor Biochemie
Klíčová slova microfluidic; haloalkane dehalogenase DbjA
Popis Analysis of concentration-dependencies is key for the quantitative understanding of biological and chemical systems. In experimental tests involving concentration gradients such as inhibitor library screening, the number of data points and the ratio between the stock volume and the volume required in each test determine the quality and efficiency of the information gained. Titerplate assays are currently the most widely used format, even though they require microlitre volumes. Compartmentalization of reactions in pico- to nanolitre water-in-oil droplets in microfluidic devices provides a solution for massive volume reduction. This work addresses the challenge to produce microfluidic-based concentration gradients in a way that every droplet represents one unique reagent combination. We present a simple microcapillary technique able to generate such series of monodisperse water-in-oil droplets (with frequency up to 10 Hz) from a sample presented in an open well, e.g. a titreplate. Time-dependent variation of the well content results in microdroplets that represent time capsules of the composition of the source well.
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