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      Continuous-flow sorting of microalgae cells based on lipid content by high frequency dielectrophoresis

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          Abstract

          This paper presents a continuous-flow cell screening device to isolate and separate microalgae cells ( Chlamydomonas reinhardtii) based on lipid content using high frequency (50MHz) dielectrophoresis. This device enables screening of microalgae due to the balance between lateral DEP forces relative to hydrodynamic forces. Positive DEP force along with amplitude-modulated electric field exerted on the cells flowing over the planar interdigitated electrodes, manipulated low-lipid cell trajectories in a zigzag pattern. Theoretical modelling confirmed cell trajectories during sorting. Separation quantification and sensitivity analysis were conducted with time-course experiments and collected samples were analysed by flow cytometry. Experimental testing with nitrogen starved dw15-1 (high-lipid, HL) and pgd1 mutant (low-lipid, LL) strains were carried out at different time periods, and clear separation of the two populations was achieved. Experimental results demonstrated that three populations were produced during nitrogen starvation: HL, LL and low-chlorophyll (LC) populations. Presence of the LC population can affect the binary separation performance. The continuous-flow micro-separator can separate 74% of the HL and 75% of the LL out of the starting sample using a 50 MHz, 30 voltages peak-to-peak AC electric field at Day 6 of the nitrogen starvation. The separation occurred between LL (low-lipid: 86.1% at Outlet # 1) and LC (88.8% at Outlet # 2) at Day 9 of the nitrogen starvation. This device can be used for onsite monitoring; therefore, it has the potential to reduce biofuel production costs.

          Most cited references55

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          Biodiesel from microalgae beats bioethanol.

          Renewable biofuels are needed to displace petroleum-derived transport fuels, which contribute to global warming and are of limited availability. Biodiesel and bioethanol are the two potential renewable fuels that have attracted the most attention. As demonstrated here, biodiesel and bioethanol produced from agricultural crops using existing methods cannot sustainably replace fossil-based transport fuels, but there is an alternative. Biodiesel from microalgae seems to be the only renewable biofuel that has the potential to completely displace petroleum-derived transport fuels without adversely affecting supply of food and other crop products. Most productive oil crops, such as oil palm, do not come close to microalgae in being able to sustainably provide the necessary amounts of biodiesel. Similarly, bioethanol from sugarcane is no match for microalgal biodiesel.
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            Cellular dielectrophoresis: applications to the characterization, manipulation, separation and patterning of cells.

            Over the past decade, dielectrophoresis (DEP) has evolved into a powerful, robust and flexible method for cellular characterization, manipulation, separation and cell patterning. It is a field with widely varying disciplines, as it is quite common to see DEP integrated with a host of applications including microfluidics, impedance spectroscopy, tissue engineering, real-time PCR, immunoassays, stem-cell characterization, gene transfection and electroporation, just to name a few. The field is finally at the point where analytical and numerical polarization models can be used to adequately describe and characterize the dielectrophoretic behavior of cells, and there is ever increasing evidence demonstrating that electric fields can safely be used to manipulate cells without harm. As such, DEP is slowly making its way into the biological sciences. Today, DEP is being used to manipulate individual cells to specific regions of space for single-cell assays. DEP is able to separate rare cells from a heterogeneous cell suspension, where isolated cells can then be characterized and dynamically studied using nothing more than electric fields. However, there is need for a critical report to integrate the many new features of DEP for cellular applications. Here, a review of the basic theory and current applications of DEP, specifically for cells, is presented. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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              Strategies for dielectrophoretic separation in laboratory-on-a-chip systems.

              Dielectrophoresis--the induced motion of polarizable particles in nonuniform fields--has proven to be an effective method for the separation of bioparticles such as cells, viruses and proteins. In this paper, the application of dielectrophoresis in miniaturized laboratories-on-a-chip is discussed, and strategies are described for using dielectrophoresis both for the binary separation of bioparticles and for the fractionation of many populations in such devices.
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                Author and article information

                Contributors
                Journal
                AIMS Biophysics
                AIMS Biophysics
                AIMS Press
                2377-9098
                29 August 2016
                : 3
                : 3
                : 398-414
                Affiliations
                [1 ] Mechanical Engineering Department, University of Nevada, Reno, USA
                [2 ] Physiology and Cell Biology Molecular Bioscience, University of Nevada, School of Medicine, Reno, USA
                [3 ] Civil and Environmental Engineering Department, University of Nevada, Reno, USA
                Author notes
                Emil J. Geiger, E-mail: ejg@ 123456unr.edu ; Tel: +775-784-6931; Fax: +775-784-1701.
                Article
                10.3934/biophy.2016.3.398
                1692c3a3-9007-447a-873f-215900ee3b95
                Copyright © 2016 by AIMS Environmental Science

                This work is licensed under a Creative Commons Attribution-NonCommercial-Share Alike 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/

                CC BY 3.0

                History
                : 18 July 2016
                : 21 August 2016
                Categories
                Research article

                Biophysics
                Cell sorting,dielectrophoresis,biofuel,microalgae,flow cytometry
                Biophysics
                Cell sorting, dielectrophoresis, biofuel, microalgae, flow cytometry

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