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      A flow through device for simultaneous dielectrophoretic cell trapping and AC electroporation

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          Abstract

          Isolation of cells and their transfection in a controlled manner is an integral step in cell biotechnology. Electric field approaches such as dielectrophoresis (DEP) offers a more viable method for targeted immobilization of cells without any labels. For transfection of cells to incorporate exogenous materials, electrical methods such as electroporation, are preferred over chemical and viral delivery methods since they minimally affect cell viability and can target many types. However prior approaches to both methods required multiple excitation sources, an AC source for DEP-based trapping and another DC source for electroporation. In this paper, we present a first of its kind flow through lab-on-chip platform using a single AC excitation source for combined trapping using negative dielectrophoresis (nDEP) and AC electroporation. Use of AC fields for electroporation eliminates the unwanted side effects of electrolysis or joule heating at electrodes compared to DC electroporation. Adjusting the flow rate and the electrical parameters of the incident AC field precisely controls the operation (trap, trap with electroporation and release). The platform has been validated through trapping and simultaneous transfection of HEK-293 embryonic kidney cells with a plasmid vector containing a fluorescent protein tag. Numerical scaling analysis is provided that indicates promise for individual cell trapping and electroporation using low voltage AC fields.

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          Slow viscous motion of a sphere parallel to a plane wall—I Motion through a quiescent fluid

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            Theory of Nanometric Optical Tweezers

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              Free flow acoustophoresis: microfluidic-based mode of particle and cell separation.

              A novel method, free flow acoustophoresis (FFA), capable of continuous separation of mixed particle suspensions into multiple outlet fractions is presented. Acoustic forces are utilized to separate particles based on their size and density. The method is shown to be suitable for both biological and nonbiological suspended particles. The microfluidic separation chips were fabricated using conventional microfabrication methods. Particle separation was accomplished by combining laminar flow with the axial acoustic primary radiation force in an ultrasonic standing wave field. Dissimilar suspended particles flowing through the 350-microm-wide channel were thereby laterally translated to different regions of the laminar flow profile, which was split into multiple outlets for continuous fraction collection. Using four outlets, a mixture of 2-, 5-, 8-, and 10-microm polystyrene particles was separated with between 62 and 94% of each particle size ending up in separate fractions. Using three outlets and three particle sizes (3, 7, and 10 microm) the corresponding results ranged between 76 and 96%. It was also proven possible to separate normally acoustically inseparable particle types by manipulating the density of the suspending medium with cesium chloride. The medium manipulation, in combination with FFA, was further used to enable the fractionation of red cells, platelets, and leukocytes. The results show that free flow acoustophoresis can be used to perform complex separation tasks, thereby offering an alternative to expensive and time-consuming methods currently in use.
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                Author and article information

                Contributors
                sameer@ece.tufts.edu
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                19 August 2019
                19 August 2019
                2019
                : 9
                : 11988
                Affiliations
                [1 ]ISNI 0000 0004 1936 7531, GRID grid.429997.8, Tufts University, Department of Electrical and Computer Engineering, ; 161 College Ave, Medford, MA 02155 USA
                [2 ]Nano Lab, Advanced Technology Laboratory, 200 Boston Ave, Medford, MA 02155 USA
                [3 ]ISNI 0000 0004 1936 7531, GRID grid.429997.8, Allen Discovery Center at Tufts University, Department of Biology, ; Medford, MA 02155 USA
                Author information
                http://orcid.org/0000-0002-4384-3476
                http://orcid.org/0000-0001-7292-8084
                Article
                48198
                10.1038/s41598-019-48198-x
                6700080
                31427614
                fd28b07b-6c1b-40db-adb5-fde6ec156b62
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 30 January 2019
                : 30 July 2019
                Funding
                Funded by: FundRef https://doi.org/10.13039/100000001, National Science Foundation (NSF);
                Award ID: CBET-0939511
                Award ID: EEC-1444926
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/100007297, United States Department of Defense | United States Navy | ONR | Office of Naval Research Global (ONR Global);
                Award ID: N00014-16-1-2550
                Award ID: N00014-12-1-0888
                Award Recipient :
                Categories
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                © The Author(s) 2019

                Uncategorized
                biotechnology,engineering
                Uncategorized
                biotechnology, engineering

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