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AC electrokinetic characterisation and separation of cells with high and low embryogenic potential in suspension cultures of carrot (Daucus carota)

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Abstract

Suspension cultures of Daucus carota L. were established, and cells with embryogenic potential were separated from those without by density gradient centrifugation in Ficoll at different stages in the growth curve. In order to obtain information about the electrical properties of individual cells, electrorotation spectra of single plant cells from different fractions were measured before and after induction of embryogenesis. The data were analysed using models based on Maxwell–Wagner's theories of interfacial polarisation. It was found that the denser cells had a higher embryogenic potential, a darker appearance and a higher internal conductivity (>1 S m−1) than the less dense cells, which had less or no embryogenic potential and a lower internal conductivity (<1 S m−1). Modelling the dielectrophoretic (DEP) response on the basis of the electrorotation data suggested that separation of cells with high embryogenic potential may be achievable in the frequency range 1–10 MHz. Actual dielectrophoretic separation of cells with high embryogenic potential from suspensions in which embryogenesis had not yet been induced was achieved using steric as well as hyperlayer dielectrophoretic Field-Flow Fractionation (DEP-FFF).

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References

  • Arnold WM & Zimmermann U (1988) Electrorotation: development of a technique for dielectric measurements on individual cells and particles. J. Electrostat. 21: 151–191

    Google Scholar 

  • Arnold WM, Zimmermann U, Pauli W, Benzig M, Niers C & Ahlers J (1988) The comparative influence of substituted phenols (especially chlorophenols) on yeast cells assayed by electrorotation and other methods. Biochim. Biophys. Acta 942: 83–95

    Google Scholar 

  • Asami K & Yamaguchi T (1992) Dielectric spectroscopy of plant protoplasts. 63: 1493–1499

    Google Scholar 

  • Burt JPH, Pethig R, Gascoyne PRC & Becker FF (1990) Dielectrophoretic characterisation of Friend murine erythroleukaemic cells as a measure of induced differentiation. Biochim. Biophys. Acta 1034: 93–101

    Google Scholar 

  • Dalton C, Goater AD, Drysdale J & Pethig R (2001) Parasite viability by electrorotation. Coll. Surf. A: Physicochem. Eng. Asp. 195: 263–268

    Google Scholar 

  • Dopp E, Jonas L, Nebe B, Budde A & Knippel E (2000) Dielectric changes in membrane properties and cell interiors of human mesothelial cells in vitro after crocidolite asbestos exposure. Environmental Health Perspectives 108: 153–158

    Google Scholar 

  • Fuhr G, Müller T, Wagner A & Donath E (1987) Electrorotation of oat protoplasts before and after fusion. Plant Cell Physiol. 28: 549–555

    Google Scholar 

  • Fuhr G, Schnelle T, Hagedorn R & Shirley SG (1995) Dielectrophoretic field cages: technique for cell, virus and macro-molecule handling. Cell. Eng. Inc. Mol. Eng. 1: 47–57

    Google Scholar 

  • Fujimura T & Komamine A (1979) Synchronization of somatic embryogenesis in a carrot cell suspension culture. Plant Physiol. 64: 162–164

    Google Scholar 

  • Fujimura T & Komamine A (1980) Aspects of DNA, RNA and protein synthesis during somatic embryogenesis in carrot cell suspension cultures. Physiol. Plant 49: 255–260

    Google Scholar 

  • Fujimura T & Komamine A (1981) Changes in chromosomal proteins during early changes of synchronized embryogenesis in a carrot cell suspension culture. Z. Pflanzenphysiol. 102: 293–298

    Google Scholar 

  • Glaser R, Fuhr G, Gimsa J & Hagedorn R (1985) Electrorotation: capabilities and limitations. Studia Biophys. 110: 43–50

    Google Scholar 

  • Gascoyne PRC, Becker FF & Wang XB (1995) Numerical analysis of the influence of experimental conditions on the accuracy of dielectric parameters derived from electrorotation measurements. Bioelectrochem. Bioenerg. 36: 115–125

    Google Scholar 

  • Gimsa J, Fuhr G & Glaser R (1985) Interpretation of electrorotation of protoplasts. Studia Biophys. 109: 5–14

    Google Scholar 

  • Gimsa J, Müller T, Schnelle T & Fuhr G (1996) Dielectric spectroscopy of single human erythrocytes at physiological ionic strength: dispersion of the cytoplasm. Biophys. J. 71: 495–506

    Google Scholar 

  • Goater AD & Pethig R (1998) Electrorotation and dielectrophoresis. Parasitology 117: s177–s189

    Google Scholar 

  • Goldsworthy A & Mina MG (1991) Electrical patterns of tobacco cells in media containing indole-3-acetic acid or 2,4-dichloro-phenoxyacetic acid - their relation to organogenesis and herbicide action. Planta 183: 368–373

    Google Scholar 

  • Huang Y, Holzel R, Pethig R & Wang XB (1992) Differences in the AC electrodynamics of viable and non-viable yeast cells de-termined through combined dielectrophoresis and electrorotation studies. Phys. Med. Biol. 37: 1499–1517

    Google Scholar 

  • Hughes MP (2000) AC electrokinetics: applications for nanotechnology. Nanotechnology 11: 124–132

    Google Scholar 

  • Jones TB (1995) Electromechanics of Particles. Cambridge University Press, Cambridge

    Google Scholar 

  • Kakutani T, Shibatani S & Senda M (1993) Electrorotation of barley mesophyll protoplasts. Bioelectrochem. Bioenerg. 31: 85–97

    Google Scholar 

  • Markx GH & Davey CL (1999) The dielectric properties of biological cells at radio-frequencies: applications in biotechnology. Enzyme Microb. Technol. 25: 161–171

    Google Scholar 

  • Markx GH & Pethig R (1995) Dielectrophoretic separation of cells: continuous separation. Biotechnol. Bioeng. 45: 337–343

    Google Scholar 

  • Markx GH & Preil W(1991) Dielectric spectroscopy of plant cell and somatic embryo cultures. 1991 IAPTC Tagung der deutschen Sektion. IAPTC, Hamburg, Germany (p. 55)

    Google Scholar 

  • Markx GH, Pethig R & Rousselet J (1997a) The dielectrophoretic levitation of latex beads, with reference to field-flow fractiona-tion. J. Phys. D: Appl. Phys. 30: 2470–2477

    Google Scholar 

  • Markx GH, Rousselet J & Pethig R (1997b) DEP-FFF: Field-Flow Fractionation using non-uniform fields. J. Liq. Chrom. Rel. Technol. 20: 2857–2872

    Google Scholar 

  • Nomura K & Komamine A (1985) Identification and isolation of single cells that produce somatic embryos at high frequency in a carrot suspension culture. Plant Physiol. 79: 988–991

    Google Scholar 

  • Osuga K & Komamine A (1994) Synchronization of somatic embryogenesis from carrot cells at high frequency as basis for mass production of embryos. Plant Cell Tiss. Org. Cult. 39: 125–135

    Google Scholar 

  • Pohl HA (1978) Dielectrophoresis. Cambridge University Press, Cambridge

    Google Scholar 

  • Rathore KS, Hodges TK & Robinson KR (1988) Ionic basis of currents in somatic embryos of Daucus carota. Planta 175: 280–289

    Google Scholar 

  • Sukhorukov VL, Benkert R, Obermeyer G, Bentrup FW & Zimmermann U (1998) Electrorotation of isolated generative and vegetative cells, and of intact pollen grains of Lilium longiflorum. J. Membrane Biol. 161: 21–32

    Google Scholar 

  • Toonen MAJ, Hendriks T, Schmidt EDL, Verhoeven HA, van Kammen A & deVries SC (1994) Description of somatic embryo forming cells in carrot suspension cultures employing video cell tracking. Planta 194: 565–572

    Google Scholar 

  • Tsukahara M & Komamine A (1997) Separation and analysis of cell types involved in early stages of carrot somatic embryo-genesis. Plant Cell Tiss. Org. Cult. 47: 145–151

    Google Scholar 

  • Vasil IK (ed) (1984) Cell Culture and Somatic Cell Genetics of Plants. Academic Press, Orlando, Florida

    Google Scholar 

  • Wang XB, Huang Y, Gascoyne PRC, Becker FF, Hölzel R & Pethig R (1994) Changes in Friend murine erythroleukemia cell-mem-branes during induced differentiation determined by electrorota-tion. Biochim. Biophys. Acta 1193: 330–344

    Google Scholar 

  • Wang XB, Yang J, Huang Y, Vykoukal J, Becker FF & Gascoyne PRC (2000) Cell separation by dielectrophoretic field-flow-fractionation. Anal. Chem. 72: 832–839

    Google Scholar 

  • Wurtele ES, Keller GL, Nikolau BJ & Ulrich TH (1988) Quantitation of starch and ADP-glucose pyrophosphorylase in non-embryogenic cells and embryogenic cell clusters from carrot sus-pension cultures. J. Plant Physiol. 132: 683–689

    Google Scholar 

  • Zhou XF, Markx GH & Pethig R (1996) Effect of biocide con-centration on electrorotation spectra of yeast cells. Biochim. Biophys. Acta 128: 60–64

    Google Scholar 

Download references

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Correspondence to Gerard H. Markx.

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Falokun, C.D., Mavituna, F. & Markx, G.H. AC electrokinetic characterisation and separation of cells with high and low embryogenic potential in suspension cultures of carrot (Daucus carota). Plant Cell, Tissue and Organ Culture 75, 261–272 (2003). https://doi.org/10.1023/A:1025860601205

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