Biochimica et Biophysica Acta 1721 (2005) 130 – 138

http://www.elsevier.com/locate/bba

Boundary-element calculations for dielectric behavior of

doublet-shaped cells
Katsuhisa Sekinea,*, Yoko Watanabea, Saori Haraa, Koji Asamib
a
School of Health Sciences, Faculty of Medicine, Kanazawa University, 5-11-80 Kodatsuno, Kanazawa, 920-0942, Japan
b
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan

Received 25 February 2004; accepted 18 October 2004

Available online 30 October 2004

Abstract

In order to simulate dielectric relaxation spectra (DRS) of budding yeast cells (Saccharomyces cerevisiae) in suspension, the complex
polarization factor (Clausius–Mossotti factor) b for a single cell and the complex permittivity of a cell suspension e*sus were calculated with a
doublet-shaped model (model RD), in which two spheres were connected with a part of a ring torus, using the boundary element method. The
b values were represented by a diagonal tensor consisting of components b z parallel to the rotation axis (z axis) and b h in a plane (h plane)
perpendicular to the axis. The e*sus values were calculated from the complex permittivity of the suspending medium e*a and the components of
b. The calculation was compared with that of a conventional prolate spheroid model (model CP). It was found that model CP could be used
as a first approximation to model RD. However, differences existed in b z between models RD and CP; b z showed three relaxation terms in
the case of model RD in contrast with two terms in model CP. Narrowing the junction between the two spheres in model RD markedly
decreased the characteristic frequency of one of the relaxation terms in b z. This suggests that the structure of the junction can be estimated
from DRS. Effects of the shape change from model RD to a two-sphere model (model RD without the junction) were also examined. The
behavior of b z in the two-sphere model, the relaxation intensity of which was much lower than model RD, was quite similar to that in a
single-sphere model. These simulations were consistent with the experimental observations of the dielectric behavior of the yeast cells during
cell cycle progression.
D 2004 Elsevier B.V. All rights reserved.

Keywords: Clausius–Mossotti factor; Complex permittivity; Dielectric relaxation; Interfacial polarization; Saccharomyces cerevisiae; Simulation

1. Introduction permittivity of the suspending medium, and e 0 is the

Biological cells in suspension are polarized by external
electric fields as a result of interfacial polarization [1–6].
The electric dipole induced in the cell is due to charge
accumulation at the boundaries between the conductive
phases (cytoplasm and suspending medium) and the
insulating phase (cell membrane). When an AC electric
field is applied, the polarizability a of the cell depends on
the frequency f of the AC field. This is conventionally
expressed by the complex polarization factor (Clausius–
Mossotti factor) b defined as b=a/(3Veae 0) [5] or b=a/
(Veae 0) [7], where V is the cell volume, e a is the relative
* Corresponding author. Fax: +81 76 234 4360.
E-mail address: sekine@kenroku.kanazawa-u.ac.jp (K. Sekine).
0304-4165/$ - see front matter D 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.bbagen.2004.10.010
permittivity of vacuum. The dielectric relaxation spectra
(DRS) that provide information pertaining to the depend-
ence of b on f have been measured in order to estimate the
structural and electrical properties of the cells [1–16].
Experimentally, DRS are measured by two different
methods: the bsuspensionQ and the bsingle-cellQ methods
[14,15,17,18]. In the bsuspensionQ method, complex per-
mittivity of cell suspensions e*sus is measured by dielectric
spectroscopy (DS), where the complex permittivity is
defined as e*=e+j/(i2pfe 0) with relative permittivity e,
electrical conductivity j, and an imaginary unit i. The
bsingle-cellQ method analyzes single cells by dielectropho-
resis (DP), electrorotation (ER), and electroorientation (EO)
techniques [5]. These techniques utilize electromechanical
effects on the cells caused by various AC fields, i.e., the DP
 K. Sekine et al. / Biochimica et Biophysica Acta 1721 (2005) 130–138 131

measures the force Fe on the cells induced by non-uniform junction, which becomes narrower and is closed during
fields, ER measures the torque Te in rotating fields, and EO septum formation in cell division. In our previous studies
measures Te that causes orientation of non-spherical cells in [33,34,46], the dielectric behavior of yeast cells with and
uniform fields. without a bud was studied using DS, and the changes in
Theoretical formulas for the interfacial polarization [19– DS were also measured for the cells in synchronized cell
25] have been devised by solving Laplace’s equation, based culture. These results were qualitatively interpreted using
on the assumption that the charges accumulate within the conventional spherical and spheroidal models. In the
regions of infinitesimal thickness at interfaces [26,27]. The present study, we reexamine the results using more realistic
simplest model of biological cells is a spherical model with cell models.
a concentric shell in which the core and shell phases
correspond to the conductive cytoplasm and the insulating
cell membrane, respectively [21]. An ellipsoidal model with 2. Models and method of calculation
a confocal shell was developed to extend the formulas to
non-spherical cells [5,22–25]. Analytical solutions were 2.1. Models
available for both the models. In the ellipsoidal model, b
transforms into a diagonal tensor that consists of three The dielectric behavior of biological cells is mainly
components along the three semiaxes of the ellipsoid. Using attributed to the inhomogeneous structure of the cells, which
the ellipsoidal model, the DRS observations can be consists of the electrically conductive cytoplasm and the
interpreted as follows: Along with the Maxwell–Wagner insulating cell membrane. In the case of yeast cells, the cell
approach [24], e sus
* was given as a function of the complex wall [35–38] and the vacuole [47,48] also contribute to the
permittivity of the suspending medium e*a and the three dielectric behavior. However, in this study, at the initial
components of b. In the single-cell method [5], Fe in DP stages of the examination, we used simple models consist-
depends on the real part of b along one of the semiaxes of ing of only the homogeneous cytoplasm and the cell
the ellipsoid when the gradient of the AC field intensity is membrane. Effects of the cell wall and the vacuole should
parallel to this axis. In ER, when the model is axially be examined in future studies, if necessary.
symmetric, the Te for its rotation around the axis of Fig. 1 shows the realistic models used in this study. In
symmetry is a function of the imaginary part of b model RD, the mother and daughter (bud) cells are
perpendicular to the axis of symmetry. The EO is represented by two spheres: one (mother cell) has a radius
determined by the differences in the real parts of the b R m centered at Z m on the z axis and the other (daughter
components. In several experimental studies, the ellipsoidal
model with a concentric shell was used to analyze
experimental data for non-spherical cells, e.g., llama
erythrocytes were represented by an ellipsoid [28], human
erythrocytes by an oblate spheroid [24,28–31], and yeast
cells by a prolate spheroid [32–34]. A prolate spheroid with
two concentric shells was used to represent the cell
membrane and the cell wall of yeast cells [35–38].
The ellipsoidal models, for which analytical equations
are available, are useful for examining the effects of the
cell shape on DRS. However, these do not necessarily
represent the real cell shape and do not meet the
requirements of uniformity in shell thickness. Hence,
theoretical examinations using models of realistic shape
are necessary for more detailed and reliable analyses. In
order to perform such examinations, several numerical
methods have been proposed [39–45]. In a previous paper
[42], we applied a method in which the boundary element
method (BEM) was employed to rod-shaped models with a
shell of uniform thickness relevant to the fission yeast
Schizosaccharomyces pombe. Numerical calculation with
these models provided a relaxation term that could not be
expected from the conventional ellipsoidal models. In the
present study, this calculation method is applied to models
that correspond to the budding yeast Saccharomyces
cerevisiae. The budding yeast cells are doublet-shaped, Fig. 1. Realistic models for doublet-shaped cells. The thick lines
i.e., mother and daughter (bud) cells are connected at a surrounding the models indicate the shells.
132 K. Sekine et al. / Biochimica et Biophysica Acta 1721 (2005) 130–138

cell) has a radius R d centered at Z d. The junction between
the spheres is represented by a part of a ring torus. In
order to systematically examine the effects of the shape
using model RD, the structural parameters were varied in
two series of calculations (see Table 1). In the series RDd,
the mother cell and the junction were unchanged and the
R d was changed to simulate the growth of the daughter
cell, and in the series RDj, the radius R j of the junction
was changed to simulate the septum formation between
the mother and the daughter cells. Model RT, which
consisted of two separate identical spheres of radius R m,
was used for examining electrical interactions between the
mother and the daughter cells that were still attached after
the septum formation was completed. In this model, the
distance 2Z m between the centers of the spheres was
changed, as shown in Table 1. The thickness T of the thin
shell that surrounds models RD and RT is uniform. The
values of R m, T, and e and j of the inner, outer, and shell
phases were the same as those used in the previous study
[34]: R m=1.5 Am, T=7 nm, e i=e a=80, j i=0.3 S/m, e s=5.5,
and j s=0, where the subscripts i, a, and s denote the
inner, outer, and shell phases, respectively.
In order to compare with these realistic models,
calculations were also carried out using the conventional
spheroidal (model CP) and spherical (model CS) models.
Model CP consisted of two confocal prolate spheroids. The
semiaxis of the outer spheroid in the xy plane was same as
R m, and the semiaxis R CPz along the z axis was varied. The
inner spheroid had semiaxes of (R m 2

K)1/2 in the xy plane
2
and (R CPz
K) 1/2
along the z axis with parameter K
representing a family of confocal surfaces. The K value
was determined for providing the same volume for the inner
spheroid as a spheroid of semiaxes of R m
T in the xy plane
and R CPz
T along the z axis, namely,


1=2
R2m
K R2CPz
K ¼ ðRm
T Þ2 ðRCPz
T Þ: ð1Þ
Model CS consisted of two concentric spheres of radii
R m and R m
T.
2.2. Method of calculation
The complex polarization factor b of a cell model was
evaluated by analyzing the electric potential / around the
model, induced by the external electric field E0. When the
model particle is placed in an uniform external electric field
E0(E x0, E y0, E z0), the particle is polarized and has an
induced dipole moment p, which is represented as p=aE0
with the polarizability tensor a. The / due to p at a point
r(x, y, z) distant from the particle can be expressed as /=pr/
(4pe ae 0jrj3) [49]. Hence, a and b are determined by
analyzing / around the particle.
The values of / for the models RD and RT were
calculated using the previously reported method in which
BEM was employed [41]. In series RDd of model RD, p
was placed at (0, 0, z 0) apart from the origin of the
coordinate system because the shape was asymmetrical with
respect to the xy plane. Hence, using b defined as b=a/
(Veae 0) [7], / is expressed as:
V

/¼ h i3=2 ½bh xEx0 þ yEy0
4p x2 þ y2 þ ð z
z0 Þ2
þ bz ð z
z0 ÞEz0 ; ð2Þ

where subscripts z and h denote the directions along the z

Table 1 axis and the h plane that is perpendicular to the z axis,
Structural parameters in models RD and RT relative to the mother cell
respectively. The / value for the other models that are
radius R m
symmetrical with respect to the xy plane is expressed by Eq.
Mother Daughter Daughter Junction
cell cell cell radius
(2) with z 0=0. Preliminary examinations indicated that four
position radius position significant figures were attained in the calculations of b

Z m/R m R d/R m Z d/R m R j/R m
when the surfaces were divided into more than 360 elements
in the case of model RD and 272 elements in the case of
Model RD (doublet-shaped)
RDd04 1.100 0.4 0.336 0.3
model RT. The calculations of b for the models CP and CS
RDd05 1.100 0.5 0.496 0.3 were carried out using the analytical equations [24,21].
RDd06 1.100 0.6 0.631 0.3 Following Asami et al. [24], e *sus of suspensions in which
RDd07 1.100 0.7 0.756 0.3 the particles are randomly oriented is given by the relation
RDd08 1.100 0.8 0.874 0.3
RDd09 1.100 0.9 0.989 0.3 e4sus
e4a P
RDd10 1.100 1.0 1.100 0.3 ¼ ð2bh þ bz Þ; ð3Þ
e4sus þ 2ea4 9
RDj01 1.100 1.0 1.100 0.1
RDj02 1.100 1.0 1.100 0.2 where P is the volume fraction of the particles. When Pb1,
RDj03 (RDd10) 1.100 1.0 1.100 0.3 Eq. (3) becomes a simple relation [33]
RDj04 1.100 1.0 1.100 0.4
RDj05 1.100 1.0 1.100 0.5 4 þ e4Dz ;
ðe4sus
e4a Þ=P ¼ 2eDh ð4Þ
Model RT (two spheres without junction)
RT025 1.025 1.0 1.025 –
where
RT100 1.100 1.0 1.100 –
RT400 1.400 1.0 1.400 – e4Dk ¼ e4a bk =3; ðk ¼ z; hÞ; ð5Þ
 K. Sekine et al. / Biochimica et Biophysica Acta 1721 (2005) 130–138 133

The complex quantities bk and e*Dk are expressed using

V ) and imaginary (bUk and eDk
their real (bkV and eDk U ) parts, as
bk ¼ bkV þ ibkW; ð6Þ
L
4 ¼ eDk
eDk W þ jDk =ði2pf e0 Þ;
V
ieDk ð7Þ
L
where k Dkis the limiting value of the conductivity part of
e*Dk at low frequencies.

3. Results

3.1. Dielectric spectra of doublet-shaped and prolate

spheroid models

Fig. 2A and B shows frequency dependence of e*Dz and

e*Dh for model RDj01 in the case where j a/j i=1. The
frequency dependence of (e *sus
e*)/P
a was obtained from
e*Dz and e*Dh using Eq. (4) (Fig. 2C). For comparison, these
figures also include the spectra of model CP that has the
same size as model RDj01, i.e., R CPz/R m=2.1 (model CP21).
As shown in Fig. 2A, marked differences exist in e*Dz
between models RDj01 and CP21; two relaxation terms are
found for model RDj01, however, one term is found for
model CP21. On the other hand, in e*Dh , the behavior of
model RDj01 is the same as that of model CP21 (Fig. 2B)

Fig. 3. Frequency dependence of the real (o) and imaginary parts (.) of b z
(=b zV+ ib zU) and e 4Dz for model RDj01 under the conditions (A) j a/j i=10
3,
(B) j a/j i=1, and (C) j a/j i=102. The vertical bars indicate the characteristic
frequencies of relaxation terms zLRD, zMRD, and zHRD.

and also that of model CS (not shown in this figure), which

is spherical in shape. The e sus * for model RDj01 is
represented by two relaxation terms (Fig. 2C). The low-
frequency relaxation term corresponds to that of e*Dz . The
high-frequency term includes both the relaxation in e * Dh and
the high-frequency term in e*Dz ; the two terms overlap each
other. On the other hand, e sus
* for model CP21 appears to be
a one-step relaxation, because the relaxation terms in e*Dz
and e *Dh are located in a similar frequency region. Similar to
* , the behavior of b z differed between the models RDj01
eD
and CP21; however, no differences were found in b h.
Therefore, hereafter we confine our discussion to e*Dz and
* (=e Dk
Fig. 2. Frequency dependence of e Dk V
ie DkW +jLRD /
Dk (i2pfe 0)) for the b z.
doublet-shaped model RDj01 under the condition j a/j i=1. (A) The real e DzV
U (.) parts along the z axis; (B) e Dh U (.) in
Fig. 3 shows the spectra of b z and e*Dz for model RDj01
(o) and imaginary e Dz V (o) and e Dh
the h plane; (C) the real (o) and imaginary (.) parts of (e*sus
e*)/P under the conditions j a/j i=10
3, 1, and 102. When j a/j i=1,
a
calculated with Eq. (4). The broken curves show the spectra of the there are two relaxation terms (Fig. 3B), whereas three terms
spheroidal model CP21 (model CP that has the same size as model RDj01). appear in cases where j a/j ip1 (Fig. 3A and C). The three
134 K. Sekine et al. / Biochimica et Biophysica Acta 1721 (2005) 130–138

terms of low-, middle-, and high-frequency are denoted by Table 2

zLRD, zMRD, and zHRD, respectively. In the case of the Relaxation parameters used to represent b z and e *Dz
conventional models CP and CS, b z and e*Dz contained one Model shape RD RT (two CP (prolate CS (single
doublet spheres) spheroid) sphere)
term when j a/j i=1 and two terms when j a/j ip1. The low-
and high-frequency terms were denoted by zLCP and zHCP, Maximum number 3 2 2 2
of relaxation terms
respectively, in the case of model CP, and in the case of
model CS, they were denoted by zLCS and zHCS, Low-frequency limit
b zV bLRD bLRT bLCP bLCS
respectively. These results suggest that b z and e*Dz for e Dz
V
z
e LRD
z
e LRT
z
e LCP
z
e LCS
Dz Dz Dz Dz
model RDj01 contain one additional relaxation term that j Dz j LRD
Dz j LRT
Dz j LCP
Dz j LCS
Dz
cannot be expected from the conventional models CP and Low-frequency relaxation term
CS. The difference in the number of relaxation terms Name zLRD zLRT zLCP ZLCS
between model RD and the conventional models was found fc f czLRD f czLRT f czLCP f czLCS
in all cases examined in the present study. m m zLRDc1 m zLRTc1 m zLCP=1 m zLCS=1
For further analyses of b z and e*Dz , each of the relaxation Intermediate value
terms was characterized by assuming the Cole–Cole type b zV bM1RD
z bMRT
z bMCP
z bMCS
z

relaxation [50]. The b z and e*Dz for model RD contain a e Dz

V e M1RD
Dz e MRT
Dz e MCP
Dz e MCS
Dz

maximum of three relaxation terms zLRD, zMRD and Middle-frequency relaxation term
zHRD, Name zMRD
fc f czMRD
m m zMRDc1
bLRD
z
bM1RD
z bM1RD
z
bM2RD
z Intermediate value
bz ¼ mzLRD
þ zMRD b zV bM2RD
1 þ ðif =fczLRD Þ 1 þ ðif =fczMRD Þm z
e Dz
V e M2RD
Dz

bM2RD
bHRD High-frequency relaxation term

þ z z
zHRD þ bHRD
z ; ð8Þ Name zHRD zHRT zHCP ZHCS
1 þ ðif =fczHRD Þm fc f czHRD f czHRT f czHCP f czHCS
m m zHRDc1 m zHRTc1 m zHCP=1 m zHCS=1
High-frequency limit
jLRD eLRD
Dz
eDz
M1RD
bVz bHRD bHRT bHCP bHCS
4 ¼ Dz þ
eDz z z z z
zLRD e Dz
V e HRD e HRT e HCP e HCS
i2pf e0 1 þ ðif =fczLRD Þm Dz Dz Dz Dz

The b z and e*Dz for model RD are represented by Eqs. (8) and (9). Similar
eM1RD
Dz
eM2RD
Dz relations containing two relaxation terms are used in order to represent the
þ zMRD b z and e*Dz for the other models.
1 þ ðif =fczMRD Þm

eM2RD
eHRD
þ Dz Dz
zHRD þ eHRD
Dz ;
1 þ ðif =fczHRD Þm
where f c is the characteristic frequency, m is the Cole–
Cole parameter, and the superscripts LRD, M1RD,
M2RD, and HRD refer to the values of b zV and e Dz V at
the low-frequency limit, between zLRD and zMRD,
between zMRD and zHRD, and at the high-frequency
limit, respectively. These relaxation parameters are
listed in Table 2. Values of m were close to 1 in
most of the cases examined in the present study. In
the cases of the conventional models CP and CS, b z
and e*Dz were represented by relations similar to Eqs.
(8) and (9) that contained two relaxation terms of the
Debye type (m=1), using the relaxation parameters shown in
Table 2.
3.2. Effects of external conductivity on relaxation
parameters
As shown in Fig. 3, the relaxation spectra of b z and e*Dz
depend considerably on the external conductivity j a.
Subsequently, we will examine the effects of j a on the
ð9Þ
relaxation parameters. This examination enables us to
compare model RD with the conventional models CP and
CS in detail. In this calculation, we used models RDj01,
CP21, and CS.
Fig. 4 shows the j a dependence of the relaxation
parameters for e*Dz . This figure suggests that, irrespective
of the difference in the number of relaxation terms between
model RD and the conventional models, several similarities
exist in their dielectric behavior; for example, as shown in
Fig. 4A, f czHRDcf czHCPcf czHCS over the entire j a/j i range.
In the case when j a/j ib10
2, f czLRDcf czLCPcf czLCS.
Further, when j a/j iz10
1, f czMRDcf czLCS. In Fig. 4B, the
values of eLRD HRD
Dz and e Dz
M1RD
over the entire j a/j i range, e Dz

3 M2RD
1
at j a/j iV10 and e Dz at j a/j iz10 , agree with the
relaxation parameters for model CP or those for both model
CP and model CS. Such similarities were also found in the
behavior of b z.
Table 3 shows the relation between the relaxation
parameters for model RD and those for the two conventional
models, i.e., model CP that has the same size as model RD
and the spherical model CS, derived from Fig. 4 and other
results obtained in the present study. As shown in this table,
 K. Sekine et al. / Biochimica et Biophysica Acta 1721 (2005) 130–138 135

Table 3
Relation between the relaxation parameters for model RD and those for the
two conventional models (model CP that has the same size as model RD
and the spherical model CS)
j a/j ib1 j a/j i=1 j a/j iN1
f czLRD cf czLCP* dependent on R j dependent on R j
f czMRD dependent on R j cf czLCS cf czLCS
f czHRD cf czHCP* not found cf czHCP*
e LRD LRD
Dz , b z ce LCP LCP
Dz , b z ce LCP LCP
Dz , b z ce LCP LCP
Dz , b z

e M1RD
Dz , bM1RD
z ce MCP MCP
Dz , b z dependent on R j dependent on R j
e M2RD
Dz , bM2RD
z dependent on R j ce HRD
Dz , bHRD
z ce MCP MCP
Dz , b z

e HRD HRD
Dz , b z ce HCP HCP
Dz , b z * ce HCP HCP
Dz , b z * ce HCP HCP
Dz , b z *
Effects of the junction radius R j are also noted in this table.
* Models CP and CS provide a similar value for the relaxation
parameter.

when j a is of the same order or higher than j i (behavior of

f czMRD at j a/j iz10
1 is shown in Fig. 4A). As noted in
Table 3, effects of the junction radius R j on the relaxation
parameters for model RD were found in the present study.
These will be discussed in Section 3.4.
According to the data shown in Fig. 4B, the j a
M2RD MCP
dependence of e Dz agrees qualitatively with that of e Dz ,
M2RD
irrespective of the R j dependence of e Dz in the low-j a
MCP HCP
region (j a/j ib1 in Table 3). Since e Dz ce Dz at j a/j i=1,
M2RD HRD
e Dz is also expected to be close to e Dz at this j a/j i
value. Hence, the intensity of the relaxation-zHRD given as

Fig. 4. Effects of j a on the relaxation parameters in e*Dz for model RDj01

(o, 4, 5, and 5), model CP21 (solid lines), and model CS (broken lines).

the values f czHRD, eLRD LRD

Dz , b z , eHRD
Dz , and b z
HRD
for model
zHCP LCP
RD are approximately the same as f c , e Dz , b zLCP, eHCP
Dz ,
and b zHCP for model CP, respectively, over the entire j a
range. In model CP, R CPz did not cause marked effects on
f czHCP, e Dz
HCP
, and b zHCP. Hence, f czHRD, e Dz
HRD
and b zHRD can Fig. 5. Effects of the daughter cell radius R d on b z and e*Dz examined
zHCS using the series RDd of model RD under the condition j a/j i=1 (4 and
also be approximated by f c , e Dz , and b zHCS, respec-
HCS
5). Effects of the length along the z axis R CPz on model CP (solid lines)
tively. Relations among the other relaxation parameters are also shown for comparison. The axial ratio q in the abscissa is defined
depend on j a, as shown in Table 3. It is notable that model as a ratio of the size of the models along the z axis to the mother cell
CS provides better approximations to f czMRD than model CP diameter 2R m.
136 K. Sekine et al. / Biochimica et Biophysica Acta 1721 (2005) 130–138

M2RD HRD Table 4

e Dz
e Dz is too weak to be found in the DRS. This
situation is also represented in Table 3. Relaxation parameters for model RT under the condition j a/j i=1
Model RT025 RT100 RT400 CS
3.3. Effects of the size of daughter (bud) cell
Z m/R m 1.025 1.1 1.4
f CzLRT/MHz 3.19 3.15 3.04 2.92
In order to simulate the dielectric behavior of yeast cells
bLRT 1.360 1.381 1.436 1.500
z
while their daughter cells are growing, the daughter cell
radius R d in model RD was changed. The calculations were
100bHRT
z 6.459 6.465 6.478 6.888
carried out using the structural parameters in series RDd of e LRT
Dz 694 715 772 840
model RD under the condition j a/j i=1. In Fig. 5, the
e HRT 1.722 1.724 1.727 1.836
Dz
relaxation parameters are plotted against the axial ratio q,
which is defined as the ratio of the size along the z axis to
10j LRT
Dz /(S/m) 1.360 1.381 1.436 1.500
the diameter of the mother cell (2R m). The f czMRD is Values of the relevant parameters for model CS are also shown for
independent of q and is same as f czLCS ( f czLCP at q=1 in Fig. comparison.
5A). The behavior of b zLRD and e DzLRD is same as that of
radius R j in order to understand the dielectric behavior of
b zLCP and e LCP
Dz (Fig. 5B and C). These results suggest that
cells during cell division. The calculations were carried out
Table 3 is also valid in the case R dpR m. With respect to the
using the structural parameters in series RDj of model RD
relaxation parameters dependent on R j, f czLRD decreases
under the conditions j a/j i=10
3, 1, and 102. They showed
with an increase in q, similar to f czLCP (Fig. 5A). However,
that R j affected the relaxation parameters and that its effects
the slope of f czLRD for q is steeper than that of f czLCP. The
were dependent on j a, as shown in Table 3. The effects of R j
relaxation parameters that refer to M1RD (b zM1RD in Fig. 5B
M1RD are shown in Fig. 6. The characteristic frequencies, f czMRD at
and e Dz in Fig. 5C) and HRD (b zHRD and e Dz HRD
not
j a/j i=10
3 and f czLRD at j a/j i=1 and 102, all decreased
shown) are almost independent of q.
linearly with decreasing R j/R m. The relaxation parameters
that refer to M2RD and M1RD showed almost linear
3.4. Effects of junction radius
relations with R j/R m.
The junction between the mother and daughter cells
3.5. Dielectric behavior of two attached cells
narrows during the septum formation in cell division.
Hence, we examined the effects of the change in junction
After completion of septum formation, the mother and
daughter cells remain attached for some time. Thus, it is
interesting to examine whether the dielectric behavior of the
two attached cells differs from that of completely separated
cells. In this calculation, we used model RT in which two
spheres are closely placed, and the distance between the
spheres was changed by varying the Z m/R m ratio (see Fig. 1
and Table 1), under the condition j a/j i=1. In order to
examine the effects of j a, additional calculations were

Table 5
Relaxation parameters for model RT100 under the conditions j a/j i=10
2
and j a/j i=102
j a/j i 10
2 102
Model RT100 CS RT100 CS
f CzLRT/MHz 0.104 0.080 4.28 4.27
f CzHRT/MHz 20.4 24.7 4.97103 4.62103

bLRT
z 1.381 1.500 1.381 1.500

bMRT
z
2.904
2.407 1.364 1.480

100bHRT
z 6.465 6.888 6.465 6.888

e LRT
Dz 715 840 715 840
e MRT
Dz 74.7 62.4 10.78 15.48

e HRT
Dz 1.724 1.836 1.724 1.836
Fig. 6. Effects of the junction radius R j on the relaxation parameters Values of the relevant parameters for model CS are also shown for
examined using the series RDj of model RD. comparison.
 K. Sekine et al. / Biochimica et Biophysica Acta 1721 (2005) 130–138
carried out under the conditions j a/j i=10
2 and 102 by
keeping Z m/R m unchanged (Z m/R m=
1.1). The spectra of
b z and e*Dz for model RT were similar to those for model CS,
and were represented by the sum of two relaxation terms,
using the relaxation parameters listed in Table 2. Tables 4
and 5 show values of the relaxation parameters. These tables
show that the spectra of b z and e*Dz for model RT are
approximately the same as those for model CS.
It is observed from Table 4 that b zLCSb0 and b zHCSb0.
This means that the external electric field E 0 causes the
induced dipole moment of the shelled sphere in the direction
opposite to E0 at the low- and the high-frequency limits
under the condition j a/j i=1. In the calculations of b z and e*Dz
for model RT, the two shelled spheres were aligned parallel
to E0. Hence, in these calculations under the condition j a/
j i=1, the dipole moment of one of the spheres depressed the
effects of E0 on the adjacent sphere. As shown in Table 4,
intensities of b zLRT and b zHRT for model RT are smaller than
those of b zLCS and b zHCS for model CS, and the intensities of
b zLRT and b zHRT decrease with a decrease in the distance
between the spheres in model RT. These are attributable to
the interaction between the spheres in model RT. As
suggested from Table 5, the same situation occurs in b zLRT
and b zHRT at j a/j i=10
2, and b zLRT, b zMRT, and b zHRT at j a/
j i=102. On the contrary, b zMCSN0 when j a/j i=10
2 (Table
5). In this case, the intensity of b zMRT was larger than that of
b zMCS because the effects of E0 were enhanced by the
interaction between the two spheres in model RT.
4. Discussion
In a previous experimental study of the doublet shape of
S. cerevisiae [34], the complex permittivity of the cell
suspension, e*sus, showed two relaxation terms of approx-
imately the same intensity, separated from each other by
approximately one decade in frequency. This result was not
well simulated by the prolate spheroid model (model CP),
particularly with respect to the large difference in the
characteristic frequency between the two relaxation terms.
In the present paper, we have demonstrated that this
difference can be explained using the doublet-shaped model
(model RD) as illustrated in Fig. 2C, which was obtained
under the condition j a/j i=1. At this j a/j i value, the
characteristic frequency of relaxation-zLRD, f czLRD ,
decreases with a decrease in the junction radius R j (Fig.
6A). Hence, the presence of a narrow junction shifted the
low-frequency relaxation term in e*sus to a frequency region
lower than that expected from model CP.
In a synchronized cell culture of S. cerevisiae, Asami et
al. [46] found that the permittivity of the culture broth
showed periodic changes with time at low frequencies
below 0.3 MHz. In each permittivity cycle that corre-
sponds to one cell division cycle, the permittivity slowly
increased and then decreased rather rapidly. This result can
be clearly interpreted by the present calculations with the
137
doublet-shaped model (model RD) and the two-sphere
model (model RT) under the condition j a/j i=1. The
increase in the permittivity is attributable to the increase
in the permittivity value at the low-frequency limit of e Dz V ,
LRD
e Dz , which results from the increase in the daughter cell
radius R d in model RD (Fig. 5C) with the growth of the
daughter cell. The rapid decrease in the permittivity is
caused by the septum formation during which the low-
frequency relaxation changes as follows. At early stages,
the low-frequency relaxation term in e*sus shifts to lower
frequencies with narrowing of the junction, due to the
decrease in f czLRD with that in R j (Fig. 6A). Subsequently,
the intensity drastically decreases after completion of the
LRT
septum formation because e Dz for the attached mother
and daughter cells (model RT) is considerably lower than
LRD
e Dz for model RD with R d=R m (for example, compare
Table 4 and Fig. 2A).
The dependence of the relaxation parameters on the
junction radius R j (Fig. 6) indicates that DRS can be utilized
in characterizing the junction. The R j effects depend on j a.
The R j dependence of f czLRD is shown in Fig. 6A under the
conditions j a/j i=1 and 102. The f czLRD deviates from f czLCP
in the range j a/j iz10
1, as shown in Fig. 4A. These
suggest that f czLRD depends on R j when j a is of the same
order or higher than j i. Under these j a conditions,
relaxation-zLRD markedly appears in e*Dz (Figs. 3B and
C). Hence, it is possible to characterize the junction by
analyzing the characteristic frequency of the low-frequency
relaxation term in e*sus. On the other hand, when j a is
considerably lower than j i, R j affects relaxation-zMRD
( f czMRD at j a/j i=10
3 in Fig. 6A), which markedly appears
in b z (Fig. 3A). Hence, the bsingle-cellQ methods that
directly measure b under low-j a conditions are favorable
for obtaining information regarding the junction.
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