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      An intra-droplet particle switch for droplet microfluidics using bulk acoustic waves

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          Abstract

          To transfer cell- and bead-assays into droplet-based platforms typically requires the use of complex microfluidic circuits, which calls for methods to switch the direction of the encapsulated particles. We present a microfluidic chip where the combination of acoustic manipulation at two different harmonics and a trident-shaped droplet-splitter enables direction-switching of microbeads and yeast cells in droplet microfluidic circuits. At the first harmonic, the encapsulated particles exit the splitter in the center daughter droplets, while at the second harmonic, the particles exit in the side daughter droplets. This method holds promises for droplet-based assays where particle-positioning needs to be selectively controlled.

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          Droplet microfluidics.

          Shia-Yen Teh, Robert Lin, Lung-Hsin Hung … (2008)
          Droplet-based microfluidic systems have been shown to be compatible with many chemical and biological reagents and capable of performing a variety of "digital fluidic" operations that can be rendered programmable and reconfigurable. This platform has dimensional scaling benefits that have enabled controlled and rapid mixing of fluids in the droplet reactors, resulting in decreased reaction times. This, coupled with the precise generation and repeatability of droplet operations, has made the droplet-based microfluidic system a potent high throughput platform for biomedical research and applications. In addition to being used as microreactors ranging from the nano- to femtoliter range; droplet-based systems have also been used to directly synthesize particles and encapsulate many biological entities for biomedicine and biotechnology applications. This review will focus on the various droplet operations, as well as the numerous applications of the system. Due to advantages unique to droplet-based systems, this technology has the potential to provide novel solutions to today's biomedical engineering challenges for advanced diagnostics and therapeutics.
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            Ultrahigh-throughput screening in drop-based microfluidics for directed evolution.

            J J Agresti, E. Antipov, A. Abate … (2010)
            The explosive growth in our knowledge of genomes, proteomes, and metabolomes is driving ever-increasing fundamental understanding of the biochemistry of life, enabling qualitatively new studies of complex biological systems and their evolution. This knowledge also drives modern biotechnologies, such as molecular engineering and synthetic biology, which have enormous potential to address urgent problems, including developing potent new drugs and providing environmentally friendly energy. Many of these studies, however, are ultimately limited by their need for even-higher-throughput measurements of biochemical reactions. We present a general ultrahigh-throughput screening platform using drop-based microfluidics that overcomes these limitations and revolutionizes both the scale and speed of screening. We use aqueous drops dispersed in oil as picoliter-volume reaction vessels and screen them at rates of thousands per second. To demonstrate its power, we apply the system to directed evolution, identifying new mutants of the enzyme horseradish peroxidase exhibiting catalytic rates more than 10 times faster than their parent, which is already a very efficient enzyme. We exploit the ultrahigh throughput to use an initial purifying selection that removes inactive mutants; we identify approximately 100 variants comparable in activity to the parent from an initial population of approximately 10(7). After a second generation of mutagenesis and high-stringency screening, we identify several significantly improved mutants, some approaching diffusion-limited efficiency. In total, we screen approximately 10(8) individual enzyme reactions in only 10 h, using < 150 microL of total reagent volume; compared to state-of-the-art robotic screening systems, we perform the entire assay with a 1,000-fold increase in speed and a 1-million-fold reduction in cost.
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              Droplet microfluidic technology for single-cell high-throughput screening.

              Eric Brouzes, Martina Medkova, Neal Savenelli … (2009)
              We present a droplet-based microfluidic technology that enables high-throughput screening of single mammalian cells. This integrated platform allows for the encapsulation of single cells and reagents in independent aqueous microdroplets (1 pL to 10 nL volumes) dispersed in an immiscible carrier oil and enables the digital manipulation of these reactors at a very high-throughput. Here, we validate a full droplet screening workflow by conducting a droplet-based cytotoxicity screen. To perform this screen, we first developed a droplet viability assay that permits the quantitative scoring of cell viability and growth within intact droplets. Next, we demonstrated the high viability of encapsulated human monocytic U937 cells over a period of 4 days. Finally, we developed an optically-coded droplet library enabling the identification of the droplets composition during the assay read-out. Using the integrated droplet technology, we screened a drug library for its cytotoxic effect against U937 cells. Taken together our droplet microfluidic platform is modular, robust, uses no moving parts, and has a wide range of potential applications including high-throughput single-cell analyses, combinatorial screening, and facilitating small sample analyses.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Biomicrofluidics
                Journal ID (iso-abbrev): Biomicrofluidics
                Journal ID (coden): BIOMGB
                Title: Biomicrofluidics
                Publisher: AIP Publishing LLC
                ISSN (Electronic): 1932-1058
                Publication date (Electronic): 26 May 2017
                Publication date Collection: May 2017
                Publication date PMC-release: 26 May 2017
                Volume: 11
                Issue: 3
                Electronic Location Identifier: 031101
                Affiliations
                [1 ]Department Biomedical Engineering, Lund University , Lund, Sweden
                [2 ]Department Engineering Sciences, Uppsala University , Uppsala, Sweden
                [3 ]Science for Life Laboratory, Uppsala University , Uppsala, Sweden
                Author notes
                [a)]

                Author to whom correspondence should be addressed: anna.fornell@ 123456bme.lth.se

                Author information
                http://orcid.org/0000-0001-7980-376X
                http://orcid.org/0000-0002-1264-1337
                Article
                Publisher ID: 1.4984131 Publisher ID: 016703BMF Publisher-manuscript ID: FT-17137R
                DOI: 10.1063/1.4984131
                PMC ID: 5446280
                SO-VID: e4344e10-c045-47ec-8eaa-a50b50ddc5ff
                Copyright © © 2017 Author(s).
                License:

                1932-1058/2017/11(3)/031101/4

                All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 24 April 2017
                Date accepted : 13 May 2017
                Page count
                Pages: 4
                Funding
                Funded by: Swedish Research Council
                Funded by: Crafoordska Stiftelsen (Crafoord Foundation) http://dx.doi.org/10.13039/501100003173
                Funded by: Knut och Alice Wallenbergs Stiftelse (Knut and Alice Wallenberg Foundation) http://dx.doi.org/10.13039/501100004063
                Award ID: 2012.0023
                Funded by: Stiftelsen Olle Engkvist Byggmästare http://dx.doi.org/10.13039/501100004200
                Funded by: Uppsala Universitet (Uppsala University) http://dx.doi.org/10.13039/501100007051
                Funded by: National Institutes of Health http://dx.doi.org/10.13039/100000002
                Categories
                Subject: Fast Track
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