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Fluorescence detection methods for microfluidic droplet platforms

Record type: Article

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that accompany system miniaturisation. Advantages include the ability to efficiently process nano- to femoto- liter volumes of sample, facile integration of functional components, an intrinsic predisposition towards large-scale multiplexing, enhanced analytical throughput, improved control and reduced instrumental footprints.In recent years much interest has focussed on the development of droplet-based (or segmented flow) microfluidic systems and their potential as platforms in high-throughput experimentation. Here water-in-oil emulsions are made to spontaneously form in microfluidic channels as aresult of capillary instabilities between the two immiscible phases. Importantly, microdroplets of precisely defined volumes and compositions can be generated at frequencies of several kHz. Furthermore, by encapsulating reagents of interest within isolated compartments separated by a continuous immiscible phase, both sample cross-talk and dispersion (diffusion- and Taylor-based) can be eliminated, which leads to minimal cross-contamination and the ability to time analytical processes with great accuracy. Additionally, since there is no contact between the contents of the droplets and the channel walls (which are wetted by the continuous phase) absorption and loss of reagents on the channel walls is prevented. Once droplets of this kind have been generated and processed, it is necessary to extract the required analytical information. In this respect the detection method of choice should be rapid, provide high-sensitivity and low limits of detection, be applicable to a range of molecular species, be non-destructive and be able to be integrated with microfluidic devices in a facile manner. To address this need we have developed a suite of experimental tools and protocols that enable the extraction of large amounts of photophysical information from small-volume environments, and are applicable to the analysis of a wide range of physical, chemical and biological parameters. Herein two examples of these methods are presented and applied to the detection of single cells and the mapping of mixing processes inside picoliter-volume droplets. We report the entire experimental process including microfluidic chip fabrication, the optical setup and the process of droplet generation and detection.

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Citation

Casadevall i Solvas, Xavier, Niu, Xize, Leeper, Katherine, Cho, Soongwon, Chang, Soo-Ik, Edel, Joshua B. and deMello, Andrew J. (2011) Fluorescence detection methods for microfluidic droplet platforms Journal of Visualized Experiments, (58) (doi:10.3791/3437).

More information

Published date: 12 October 2011
Additional Information: The video component of this article can be found at http://www.jove.com/video/3437/
Keywords: bioengineering, droplet microfluidics, single cell assays, single molecule assays, fluorescence spectroscopy, fluorescence, lifetime Imaging
Organisations: Mechatronics

Identifiers

Local EPrints ID: 341131
URI: http://eprints.soton.ac.uk/id/eprint/341131
ISSN: 1940-087X
PURE UUID: ae44ddac-3cc7-45c5-bdd6-ad8cc6dc2b0a

Catalogue record

Date deposited: 16 Jul 2012 10:03
Last modified: 18 Jul 2017 05:38

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Contributors

Author: Xavier Casadevall i Solvas
Author: Xize Niu
Author: Katherine Leeper
Author: Soongwon Cho
Author: Soo-Ik Chang
Author: Joshua B. Edel
Author: Andrew J. deMello

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