Research article

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

  • Received: 18 July 2016 Accepted: 21 August 2016 Published: 29 August 2016
  • This paper presents a continuous-flow cell screening device to isolate and separate microalgae cells (Chlamydomonas reinhardtii) based on lipid content using high frequency (50 MHz) 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 starveddw15-1 (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

    Citation: Hanieh Hadady, Doug Redelman, Sage R. Hiibel, Emil J. Geiger. Continuous-flow sorting of microalgae cells based on lipid content by high frequency dielectrophoresis[J]. AIMS Biophysics, 2016, 3(3): 398-414. doi: 10.3934/biophy.2016.3.398

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  • This paper presents a continuous-flow cell screening device to isolate and separate microalgae cells (Chlamydomonas reinhardtii) based on lipid content using high frequency (50 MHz) 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 starveddw15-1 (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


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    [1] Hu Q, Sommerfeld M, Jarvis E, et al. (2008) Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant J 54: 621–639. doi: 10.1111/j.1365-313X.2008.03492.x
    [2] Chisti Y (2008) Biodiesel from microalgae beats bioethanol. Trends Biotechnol 26: 126–131. doi: 10.1016/j.tibtech.2007.12.002
    [3] Dismukes GC, Carrieri D, Bennette N, et al. (2008) Aquatic phototrophs: efficient alternatives to land-based crops for biofuels. Curr Opin Biotech 19: 235–240. doi: 10.1016/j.copbio.2008.05.007
    [4] Sharif H, Salleh A, Boyce AN, et al. Biodiesel Fuel Production from Algae as Renewable Energy. Am J Bioch Biotechnol 2008 4: 250–254.
    [5] Hannon M, Gimpel J, Tran M, et al. (2010) Biofuels from algae: challenges and potential. Curr Opin Biotech 21: 277–286. doi: 10.1016/j.copbio.2010.03.005
    [6] Bono MS, Ahner BA, Kirby BJ (2013) Detection of algal lipid accumulation due to nitrogen limitation via dielectric spectroscopy of Chlamydomonas reinhardtii suspensions in a coaxial transmission line sample cell. Bioresource Technol 143: 623–631. doi: 10.1016/j.biortech.2013.06.040
    [7] Deng YL, Kuo MY, Juang YJ (2014) Development of flow through dielectrophoresis microfluidic chips for biofuel production: Sorting and detection of microalgae with different lipid contents. Biomicrofluidics 8: 064120–064120. doi: 10.1063/1.4903942
    [8] Hadady H, Montiel C, Wetta D, et al. (2015) Liposomes as a model for the study of high frequency dielectrophoresis. Electrophoresis 36: 1423–1428. doi: 10.1002/elps.201400480
    [9] Hadady H, Wong JJ, Hiibel SR, et al. (2014) High frequency dielectrophoretic response of microalgae over time. Electrophoresis 35: 3533–3540. doi: 10.1002/elps.201400306
    [10] Michael KA, Hiibel SR, Geiger EJ (2014) Dependence of the dielectrophoretic upper crossover frequency on the lipid content of microalgal cells. Algal Research 6: 17–21. doi: 10.1016/j.algal.2014.08.004
    [11] Gagnon ZR (2011) Cellular dielectrophoresis: applications to the characterization, manipulation, separation and patterning of cells. Electrophoresis 32: 2466–2487. doi: 10.1002/elps.201100060
    [12] Hoettges KF (2009) Dielectrophoresis as a Cell Characterisation Tool. Microengineering in Biotechnology, Meth Mol Biol. New York. NY: Humana Press, 183–198.
    [13] Hughes MP (2002) Strategies for dielectrophoretic separation in laboratory-on-a-chip systems. Electrophoresis 23: 2569–2582.
    [14] Gagnon Z, Gordon J, Sengupta S, et al. (2008) Bovine red blood cell starvation age discrimination through a glutaraldehyde-amplified dielectrophoretic approach with buffer selection and membrane cross-linking. Electrophoresis 29: 2272–2279. doi: 10.1002/elps.200700604
    [15] Song H, Rosano JM, Wang Y, et al. (2015) Continuous-flow sorting of stem cells and differentiation products based on dielectrophoresis. Lab Chip 15: 1320–1328. doi: 10.1039/C4LC01253D
    [16] Valero A, Braschler T, Demierre N, et al. (2010) A miniaturized continuous dielectrophoretic cell sorter and its applications. Biomicrofluidics 4: 1–22.
    [17] Gossett DR, Weaver WM, Mach AJ, et al. (2010) Label-free cell separation and sorting in microfluidic systems. Anal and Bioanal Chem 397: 3249–3267. doi: 10.1007/s00216-010-3721-9
    [18] Mernier G, Piacentini N, Braschler T, et al. (2010) Continuous-flow electrical lysis device with integrated control by dielectrophoretic cell sorting. Lab Chip 10: 2077–2082. doi: 10.1039/c000977f
    [19] Vahey MD, Voldman J (2008) An Equilibrium Method for Continuous-Flow Cell Sorting Using Dielectrophoresis. Anal Chem 80: 3135–3143. doi: 10.1021/ac7020568
    [20] Hu X, Bessette PH, Qian J, et al. (2005) Marker-specific sorting of rare cells using dielectrophoresis. Pnas 102: 15757–15761. doi: 10.1073/pnas.0507719102
    [21] Wang X-B, Yang J, Huang Y, et al. (2000) Cell Separation by Dielectrophoretic Field-flow-fractionation. Anal Chem 72: 832–839. doi: 10.1021/ac990922o
    [22] Moon HS, Kwon K, Kim SI, et al. (2011) Continuous separation of breast cancer cells from blood samples using multi-orifice flow fractionation (MOFF) and dielectrophoresis (DEP). Lab Chip 11: 1118–1125. doi: 10.1039/c0lc00345j
    [23] Doh I, Cho YH (2005) A continuous cell separation chip using hydrodynamic dielectrophoresis (DEP) process. Sensor Actuat A-Phys 121: 59–65. doi: 10.1016/j.sna.2005.01.030
    [24] Yang J, Huang Y, Wang XB, et al. (1999) Cell Separation on Microfabricated Electrodes Using Dielectrophoretic/Gravitational Field-Flow Fractionation. J Am Chem Soc 71: 911–918.
    [25] Lewpiriyawong N, Yang C (2013) Dielectrophoresis Field-Flow Fractionation for Continuous-Flow Separation of Particles and Cells in Microfluidic Devices. Advances in Transport Phenomena 2011. Switzerland: Springer International Publishing, 29–62.
    [26] Pommer MS, Zhang Y, Keerthi N, et al. (2008) Dielectrophoretic separation of platelets from diluted whole blood in microfluidic channels. Electrophoresis 29: 1213–1218. doi: 10.1002/elps.200700607
    [27] Cheng IF, Chang HC, Hou D (2007) An integrated dielectrophoretic chip for continuous bioparticle filtering, focusing, sorting, trapping, and detecting. Biomicrofluidics 1: 21503–21515. doi: 10.1063/1.2723669
    [28] Piacentini N, Mernier G, Tornay R, et al. (2011) Separation of platelets from other blood cells in continuous-flow by dielectrophoresis field-flow-fractionation. Biomicrofluidics 5: 034122–034128. doi: 10.1063/1.3640045
    [29] Cima I, Yee CW, Iliescu FS, et al. (2013) Label-free isolation of circulating tumor cells in microfluidic devices: Current research and perspectives. Biomicrofluidics 7: 011810–011813. doi: 10.1063/1.4780062
    [30] Deng YL, Chang JS, Juang YJ (2013) Separation of microalgae with different lipid contents by dielectrophoresis. Bioresource Technol 135: 137–141. doi: 10.1016/j.biortech.2012.11.046
    [31] Schor AR, Buie CR (2012) Non-Invasive Sorting of Lipid Producing Microalgae With Dielectrophoresis Using Microelectrodes. ASME 2012 International Mechanical Engineering Congress and Exposition. Houston, Texas: American Society of Mechanical Engineers, 701–707.
    [32] Schor AR, Buie CR (2015) Dielectrophoretic sorting of lipid-containing microorganisms using high frequency electric fields produced by conducting post arrays. Solid-State Sensors, Actuators and Microsystems (TRANSDUCERS), 2015 Transducers - 2015 18th International Conference on. Anchorage, AK: IEEE, 1617–1620.
    [33] Broche LM, Labeed FH, Hughes MP (2005) Extraction of dielectric properties of multiple populations from dielectrophoretic collection spectrum data. Phys Med Biol 50: 2267–2274. doi: 10.1088/0031-9155/50/10/006
    [34] Han KH, Frazier AB (2008) Lateral-driven continuous dielectrophoretic microseparators for blood cells suspended in a highly conductive medium. Lab Chip 8: 1079–1086. doi: 10.1039/b802321b
    [35] Shima HC, Kwaka YK, Hanb CS, et al. (2009) Effect of a square wave on an assembly of multi-walled carbon nanotubes using AC dielectrophoresis. Physica E: Low-dimensional Systems and Nanostructures 41: 1137–1142. doi: 10.1016/j.physe.2008.12.007
    [36] Rodolfi L, Chini Zittelli G, Padovani G, et al. (2014) Microalgae for oil: Strain selection, induction of lipid synthesis and outdoor mass cultivation in a low‐cost photobioreactor. Biotechnol Bioeng 102: 100–112.
    [37] Li X, Moellering ER, Liu B, et al. (2012) A Galactoglycerolipid Lipase Is Required for Triacylglycerol Accumulation and Survival Following Nitrogen Deprivation in Chlamydomonas reinhardtii. The Plant Cell 24: 4670–4686. doi: 10.1105/tpc.112.105106
    [38] Sheehan J, Dunahay T, Benemann J, et al. (1998) Look back at the U. S. Department of Energy's Aquatic Species Program: Biodiesel from Algae. U.S. Department of Energy’s Office of Fuels Development.
    [39] Hadady H, Wong JJ, Hiibel SR, et al. (2014) Effect of Media Conductivity on High Frequency Dielectrophoretic Response. ASME 2014 International Mechanical Engineering Congress and Exposition. Montreal, Quebec, Canada: American Society of Mechanical Engineers. V010T013A006.
    [40] Hoettges KF, Hubner Y, Broche LM, et al. (2008) Dielectrophoresis-activated multiwell plate for label-free high-throughput drug assessment. Anal Chem 80: 2063–2068. doi: 10.1021/ac702083g
    [41] Cooper MS, Hardin WR, Petersen TW, et al. (2010) Visualizing "green oil" in live algal cells. J Biosci Bioeng 109: 198–201. doi: 10.1016/j.jbiosc.2009.08.004
    [42] Akimoto S, Mimuro M (2007) Application of Time‐Resolved Polarization Fluorescence Spectroscopy in the Femtosecond Range to Photosynthetic Systems. J Photoch Photobio 83: 163–170.
    [43] Xia Y, Whitesides GM (2003) SOFT LITHOGRAPHY. Annu Rev Mater Sci 28: 159–184.
    [44] Geiger EJ, Pisano AP, Svec F (2010) A Polymer-Based Microfluidic Platform Featuring On-Chip Actuated Hydrogel Valves for Disposable Applications. J Microelectromech S 19: 944–950. doi: 10.1109/JMEMS.2010.2048702
    [45] Aran K, Sasso LA, Kamdar N, et al. (2010) Irreversible, direct bonding of nanoporous polymer membranes to PDMS or glass microdevices. Lab Chip 10: 548–552. doi: 10.1039/b924816a
    [46] Gesche R, Kovacs R, Scherer J (2005) Mobile plasma activation of polymers using the plasma gun. Surf Coat Tech 200: 544–547. doi: 10.1016/j.surfcoat.2005.01.109
    [47] Hadady H, Michael KA, Geiger EJ (2014) Impedance Effects During High–Frequency Dielectrophoresis. ASME 2014 International Mechanical Engineering Congress and Exposition. Montreal, Quebec, Canada: American Society of Mechanical Engineers, V010T013A046.
    [48] Mooij PR, Stouten GR, Tamis J, et al. (2013) Survival of the fattest. Energ Environ Sci 6: 3404–3406. doi: 10.1039/c3ee42912a
    [49] Ling SH, Lam YC, Chian KS (2012) Continuous Cell Separation Using Dielectrophoresis through Asymmetric and Periodic Microelectrode Array. Anal Chem 84: 6463–6470. doi: 10.1021/ac300079q
    [50] Hadady H, Hiibel SR, Redelman D, et al. (2015) Use of a Separability parameter for the Design of a High Frequency Dielectrophoresis Cell Sorter Device. InterPACK/ICNMM2015. California.
    [51] Bono MS, Garcia RD, Sri-Jayantha DV, et al. (2015) Measurement of Lipid Accumulation in Chlorella vulgaris via Flow Cytometry and Liquid-State ?1H NMR Spectroscopy for Development of an NMR-Traceable Flow Cytometry Protocol. Plos One 10: 1–18.
    [52] Hyka P, Lickova S, P?ibyl P, et al. (2013) Flow cytometry for the development of biotechnological processes with microalgae. Biotechnol Adv 31: 2–16.
    [53] Montero MF, Aristizábal M, Reina GG (2011) Isolation of high-lipid content strains of the marine microalga Tetras. J Appl Phycol 23: 1053–1057. doi: 10.1007/s10811-010-9623-6
    [54] Velmurugan N, Sung M, Yim SS, et al. (2013) Evaluation of intracellular lipid bodies in Chlamydomonas reinhardtii strains by flow cytometry. Bioresource Technol 138: 30–37. doi: 10.1016/j.biortech.2013.03.078
    [55] Wang ZT, Ullrich N, Joo S, et al. (2009) Algal Lipid Bodies: Stress Induction, Purification, and Biochemical Characterization in Wild-Type and Starchless Chlamydomonas reinhardtii. Eukaryot Cell 8: 1856–1868. doi: 10.1128/EC.00272-09
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