Publication details
ENZYME KINETIC STUDIES IN DROPLET MICROFLUIDIC DEVICE WITHFLUORESCENCE DETECTION
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| Year of publication | 2024 |
| Type | Conference abstract |
| Citation | |
| Description | Microfluidic systems emerged as an alternative to larger scale analytical methods in effort to minimize consumption of chemicals while obtaining relevant data. Working in minuscule volumes presents new possibilities in sample manipulation as well as challenges. Basic concept of droplet microfluidics (DM) is generation of large number of droplets dispersed in an immiscible liquid, typically water-based reaction mixture in oil. Since the droplets are isolated and protected by the oil and can be individually manipulated, they can be regarded as separate microreactors. Compared to the conventional technologies, microfluidics offers several advantages such as reduced reagent consumption, shorter analyses times, and rapid production of data in given time. The "hearth" of the device is typically polydimethylsiloxane chip with integrated droplet generator, incubation channel and other functional elements, connected to precise syringe pumps. Many different detection techniques can be found in research papers to enable visualization and quantification in DM devices. In this study the setup allowed to measure the reaction rate simply by focusing on different distances in the incubation channel, corresponding to different reaction times. Chip fabrication process, inner surface modification as well as water-oil phase ratio and flow rates were optimized as all these have significant impact on uniform-sized droplets generation and repeatability. Enzymatic reaction of ß-galactosidase, which is an important enzyme used in biotechnology and bioanalysis was measured with its fluorogenic substrate in setup allowing on-line substrate concentration variation by setting corresponding flow rates of reaction mixture components. The aim is to develop convenient droplet microfluidics platform and optimize methods to perform enzyme assays with the above-mentioned advantages. |
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