ARTICLE IN PRESS

Journal of Electrostatics 63 (2005) 823830

www.elsevier.com/locate/elstat

High-gradient electric eld system for the

dielectrophoretic separation of cells
Zurina Z. Abidin, Gerard H. Markx
School of Chemical Engineering and Analytical Science, University of Manchester, Sackville Street,
P.O. Box 88, Manchester M60 1QD, UK
Available online 25 March 2005

Abstract
A high-gradient electric eld strength system, consisting of a chamber created from two
concentric cylindrical electrodes and lled with glass beads, was constructed for the study of
the dielectrophoretic separation of cells. The glass beads distort the electric eld patterns and
generate sites of high electric eld gradients, which can capture cells by dielectrophoresis. In
order to experimentally determine the efcacy of the system, yeast cells were injected into a
constant ow through the system, and the number of cells trapped measured. The effects of
changes in voltage, bead size and the size of the inner and outer electrodes on the trapping
efciency of the dielectrophoretic separation system were investigated. In addition, simulation
and electric eld analysis were carried out using FEMLAB. Results indicate that the trapping
of cells occurs by a combination of mechanical trapping and dielectrophoresis. Analysis of the
results allow predictions to be made for the optimum values of the voltage, bead size and size
of inner and outer electrode for any high-gradient system, in order to generate sufcient
electric eld gradient for dielectrophoretic cell collection whilst reducing non-specic
mechanical trapping.
r 2005 Elsevier B.V. All rights reserved.
Keywords: High-gradient electric eld; Dielectrophoresis; Cell separation; Electric eld model

Corresponding author. Tel.: +44 161 200 64394; fax: +44 161 200 64399.

E-mail address: gerard.markx@manchester.ac.uk (G.H. Markx).

0304-3886/$ - see front matter r 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.elstat.2005.03.078

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1. Introduction
Robust and improved cell separation techniques are important in many areas in
cell biology, biotechnology and others. The discovery of dielectrophoresis (DEP), the
induced motions of matter suspended in a uid in a non-uniform electric eld [1], has
provided new techniques for the separation of cells in the basis of their electrical
properties. The theory of dielectrophoresis has been described in a number of books
and reviews [24].
High-gradient electric eld strength separation (HGES) is one of the methods that
utilized the DEP concept. Here materials are placed between electrodes. When the
electrodes are energized, regions of highly non-uniform electric eld are induced
around the materials, and result in a net DEP force being exerted on any particle in
the uid. This method has shown potential in ltration and separation when Fritsche
[5] and Lin et al. [6] both developed a high-gradient dielectric separator and lter. In
later work, Wakeman et al. [7] used a HGES-DEP system for the ltration of
contaminants in oil, while Suehiro et al. [8] investigated the ability of a HGES-DEP
system to recover biological cells from wastewater. However, to date a systematic
investigation of the effect of column properties has been lacking. In this work we will
describe the study of some of the characteristics of the HGES system.

2. Materials and methods

2.1. HGES column
The HGES column consisted of coaxial cylindrical electrodes. The diameter of the
inner electrode varied between 0.6 and 2 mm, while the outer diameter varied
between 2.7 and 6 mm. The column had a length of 50 mm. Co-axial connectors were
used as the top and bottom cover to seal the column. The top co-axial connector had
a small rod in the middle for attaching and centring the inner electrode (see Fig. 1).
Connected to
voltage

Syringe for
feed flow

earthed

Small rod
for
electrode
attachment

Co axial
connector
for centering
Central electrode

Cylindrical
outer
electrode

Nylon filter to
support matrix

HGES
column
Rubber tubing
for protective
cover
(i)

Product
(ii)

Frame/cover
for bottom
area from
plastic

Fig. 1. High gradient electric eld strength column for DEP separation of cells. Top section was shown in
part (i) and bottom section in part (ii).

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825

A needle was stuck into the top plug for the inlet ow. The bottom cover had a
30 mm pore size nylon lter (Millipore, UK) as matrix support, and had a small hole
in it for the outlet ow. Glass beads were used as the matrix and obtained from
Sigmund Lindner GmbH in Germany. The glass beads had size distributions of
4070, 90150 and 150250 mm.
2.2. Cells
Saccharomyces cerevisiae yeast (unknown strain) was grown overnight in 100 ml
MYGP (malt 3 g l 1, yeast extract 3 g l 1, glucose 10 g l 1, mycological peptone
5 g l 1) broth at 35 1C in a shaker with a speed of 150 rpm. The cells were centrifuged
and washed four times using deionised water to reduce the conductivity. Cell
concentrations were measured using UV spectrophotometer (PYE Unicam, Philips;
model 8600) in a cuvette of 1 cm path length.
2.3. Experimental procedure
A standard frequency generator (Thurlby-Thandar, TG120) with a self-built highfrequency amplier was used to supply voltages up to 60 Vpk pk. One milliliter of the
yeast suspension was injected into the HGES-DEP column, and a signal with varying
voltage and a frequency of 1 MHz was applied in order to attract the yeast cells to
high electric eld regions by positive dielectrophoresis. Next, a deionized water ow
of 2 ml min 1 was fed into the chamber using a Sage pump syringe (Model 355, USA)
to wash away the cells which were not attracted. The outlet suspension was collected
for 30 min, and the cell concentration in the outlet suspension was then measured.
2.4. Electric field calculations
Calculations of electric eld strength, E, at dc conditions for the HGES system
was done using FEMLAB software version 2.3 (Comsol Ltd). The simulations were
done for glass beads size 50, 80, 120 and 200 mm at voltage and frequency of
40 Vpk pk and 1 MHz, respectively, with the inner and outer electrode diameter being
4.7 and 2.0 mm.

3. Results and discussion

Initial experiments were performed with a chamber with an inner electrode with a
diameter of 2 mm, and an outer electrode with a diameter of 4.7 mm. The voltage
was varied, while the frequency was xed at 1 MHz. The bead size was 4070 mm.
The results in Fig. 2 showed that a signicant number of cells are trapped when no
electric eld is applied. However, with increasing voltage an increasing number of
cells are trapped selectively by the electric eld. It is likely that the HGES column
acted similar to a deep bed ltration system, and that some of the cells were trapped
in the interstices between the beads.

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4.0E+06

no of cell trapped

3.5E+06
3.0E+06
2.5E+06

Mechanical trapping at 0 V
2.0E+06

40-70 micron - 52%

1.5E+06

90-150 micron - 48%

1.0E+06

150-250 micron - 44%

5.0E+05
0.0E+00
0

10

20

30

40

50

voltage (V)

Fig. 2. Cell capture in HGES system at different voltages with different bead sizes distributions. Bigger
beads give more mechanical trapping.

25.0

40-70 micron
90-150 micron

% electrical yield

20.0
150-250 micron
15.0

10.0

5.0

0.0
0

10

20

30

40

50

voltage (V)

Fig. 3. The % electrical yield in a HGES system with different bead sizes. The effect of bead size on cell
capture by the electric eld is small.

An increase in the bead size is likely to reduce the number of mechanically trapped
cells, as the pores between the beads become bigger. However, a change in the bead
size will also change the electric eld non-uniformities generated around and between
the beads, and hence affect the electrical trapping yield. To investigate this, the
previous experiments were repeated with different bead sizes distributions of 90150
mm and 150250 mm to give results as in Fig. 2. Increasing the bead sizes result in a
reduction of the mechanical ltering of cells. Comparison of the electrical trapping
yields using different bead sizes (Fig. 3) shows that changing the bead size has little
effect on the electrical yield. This result was surprising, as it could be expected that,
similar to the situation with microelectrodes [9], particles with a size of a small
number of multiples of the diameter of the particle to be captured would generate
non-uniform electric eld patterns that are most suitable.

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To further investigate the effect of particle size, a two-dimensional reconstruction

of the electric eld pattern in a column packed with different bead sizes was made
using FEMLAB. In addition, the electric eld around a single bead with no
neighbours, at a xed distance from the inner electrode surface, was calculated. The
results illustrated in Figs. 46 show that the highest electric eld strength occurs at
the contact points between the beads. With smaller beads, the region with high
electric eld strengths is relatively small. As the bead size increases the highest
electric eld strength at the contact point declines, but the electric eld strength

x10-4

Max: 2.6e+00
x104
2.5

Surface: electric field (E)

2
2

1
0

1.5

-1

-2
0.5
-3
-1.7

-1.6

-1.5

-1.4

-1.3

-1.2

-1.1

-1

-0.9

-0.8
x10-3

0
Min: 0

Fig. 4. The electric eld distributions around beads size 50 mm. Maximum of electric eld strength occurs
at the points of contacts of the beads. As the distance from the centre increases, the value of the electric
eld strength decreases.

x10-4

Max: 2.6e+00
x104
2.5

Surface: electric field (E)

2
2

1
0

1.5

-1

-2
0.5
-3
-1.7

-1.6

-1.5

-1.4

-1.3

-1.2

-1.1

-1

-0.9

-0.8
-3

0
Min: 0

x10

Fig. 5. The electric eld distributions around bead size 200 mm. The electric eld further away from the
points of contacts of the beads for this size is slightly stronger than 50 mm beads.

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Electric field calculated along
edge of a bead
electrode

3.0E+04

Electric field (V/m)

2.5E+04
2.0E+04

bead

1.5E+04
1.0E+04
5.0E+03

50
80 micron
120 micron
200 micron

0.0E+00
0.001

0.0011

0.0012

0.0013

distance from centre of electrode (m)

Fig. 6. Electric eld analysis using single bead. The smaller the beads, the greater the electric eld strength
at point of contact of beads. The nearly exponential rate of decay of the electric eld is also larger for
smaller beads as the distance from the centre increases.

30.0

% electrical yield

25.0
20.0
15.0
10.0
5.0
0.0
0

0.0005

0.001

0.0015

0.002

0.0025

inner electrode diameter (m)

Fig. 7. The electrical yield decreased with an increase in the dimension of the inner electrode diameter
at 40 V.

further away from the contact point declines less strongly. Thus, when the particle
size increases, the local electric eld may become less, but the volume in which the
cells may be captured increased. This situation is similar to that of microelectrodes of
different sizes, where an increase in electrode size and distance results in less rapidly
declining electric elds [10].
In order to investigate the effect of size of the electrodes used in the HGES, two
sets of experiments were performed: one with a xed outer electrode of 4.7 mm inner
diameter, but the diameter of the inner electrodes was varied from 0.6 to 2 mm, and
another one in which the size of the inner electrode was set at 1 mm, but the diameter
of the outer electrode was varied from 2.7 to 6 mm. The columns were lled with

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50.0
45.0

% electrical yield

40.0
35.0
30.0
25.0
20.0
15.0
10.0
5.0
0.0

0.002

0.004

0.006

0.008

outer electrode diameter (m)

Fig. 8. The electrical yield at 40 V decreased with the increase in the outer electrode diameter since electric
eld became weaker and only persisted near electrode surfaces.

4070 mm beads, and the trapping efciencies of the column were measured as
described previously. The results in Fig. 7 showed that when the outer electrode
diameter is kept constant, the larger the radius of the diameter of the inner electrode,
the smaller the yield of the HGES column. Conversely, when the diameter of the
inner electrode is kept constant, the column efciency decreased with the diameter of
the outer electrode (Fig. 8). These results can readily be explained by the changes in
the electric eld strength with changing column geometry [1].

Acknowledgements
We wish to thank Sigmund Lindler for the glass beads and Universiti Putra
Malaysia for nancial support.

References
[1]
[2]
[3]
[4]

H.A. Pohl, Dielectrophoresis, Cambridge University Press, Cambridge, 1978.

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(1997) 426432.
[5] G.R. Fritsche, Electrostatic separator removes FCC catalyst nes from decanted oil, Oil Gas J. 75
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(1982) 10031017.
[7] R. Wakeman, G. Butt, An investigation of high-gradient dielectrophoretic separation, Chem. Eng.
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[8] J. Suehiro, G. Zhou, M. Imamura, M. Hara, DEP lter for separation and recovery of biological cells
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[10] G.H. Markx, R. Pethig, J. Rousselet, The dielectrophoretic levitation of latex beads with reference to
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