MEASUREMENT 2017, Proceedings of the 11th International Conference, Smolenice, Slovakia

Development of Experimental Platform for Investigation of Biological

Response of Cells to Weak Low Frequency Electromagnetic Fields
1
M. Teplan, 1I. Bajla, 2O. trbk, 3M. Cifra
1
Institute of Measurement Science, Slovak Academy of Sciences, Bratislava, Slovakia
2
Biomedical Center, Jessenius Faculty of Medicine, Comenius University, Martin,
Slovakia
3
Institute of Photonics and Electronics, Czech Academy of Sciences, Prague, Czechia
Email: michal.teplan@savba.sk

Abstract. The topic of our research is based on the perspective of electromagnetic principles
of living system operation. The overall objective of this research is to expand knowledge of
the impact of weak external low-frequency electromagnetic fields (EMF) on the functioning of
selected biological objects. The particular aim of the research is to design a new
experimental approach enabling efficient scanning through EMF parameters while searching
for a specific response of the investigated biosystem. Methods based on the impedance
spectroscopy are being developed to determine the response in the growth rate of cell cultures
in an aqueous medium as well as in electrical characterization of cell structures. For this
purpose, a complex experimental platform for quantitative characterization of the biological
response to specific parameters of weak low-frequency electromagnetic fields is under
development.
Keywords: EM Fields, Biological Response, Impedance Spectroscopy

1. Introduction
Biological electromagnetic activity is a new issue for the field of biology and medicine. Since
living cells contain free and bound electric charges in the form of ions and polar molecules,
they are sensitive to EM fields and currents. Generation of endogenous EM fields was
observed in a wide frequency range (0-1015 Hz) [1]. Yet, many processes in this field have to
be clarified. It is natural to expect that the qualitative and quantitative characteristics of these
fields reflect the actual state of the organism, resulting in an enormous diagnostic and
therapeutic potential.
In our project, we focus on interactions of LF EM fields with cells. As a suitable experimental
biological model, yeast Saccharomyces cerevisiae is used. It is a eukaryotic organism that is
used in abundance for research purposes as a basic model for the eukaryotic cell, while it is
relatively easy to be cultivated, and also it has wide application in industry. The spontaneous
electrical activity of S. cerevisiae was reported in Hz, kHz, and MHz regions [2]. The
frequency-dependent biological response of proliferation was observed in the frequency range
of 1-2 kHz, with the value of magnetic induction at mT level [3].
Our main aim is to develop an experimental platform for monitoring the biological response
of cells to weak low-frequency electromagnetic fields. The key method is based on impedance
spectroscopy that appeared to be appropriate for the characterization and monitoring of cell
cultures.

2. Subject and Methods

The experimental platform comprises basic, exposure, and monitoring part. The basic part
creates a base of an experimental system with a specific architecture and spatial arrangement.

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A cultivation chamber provides suitable and stable conditions for cells growth in aqueous
media and the flow pump enables circulation of biological material (Fig. 1). The exposition
part consists of Helmholtz coils for generating electromagnetic fields together with the power
amplifier, current source, and signal generator. The monitoring part deploys impedance
spectroscopy for characterization of electrical properties of cells, turbidimetry for continuous
measurement of solution turbidity, and monitoring of basic physical and chemical conditions
of biological sample, namely temperature, pH, and solution conductivity. Moreover, an
inverted light microscope connected to a digital camera plays a role in biological sample
control and quantitative characterization of morphology and local kinetics of cells.
The measurement of the optical density of aqueous solution represents an established method
for automatic monitoring of the yeast growth curve. The approach we are developing is
focused on impedance (dielectric) spectroscopy. The problem is that the measured impedance
values depend on the conductivity of the extracellular environment, which is difficult to
control, as its electrical properties depend not only on the initial conditions of an experiment
but also on the consumption of the nutrient medium during the metabolism process of
investigated cells. This issue was partially overcome: for monitoring purposes, an estimator
for yeast density was designed, taking into account the ratio of impedance at 10 MHz and 10
kHz [4].

3. Results
SW platform enabling operation and control of the whole system together with acquisition,
pre-processing and analysis of multidimensional data is gradually built in Matlab, along with
its GUI. For optical inspection of samples, inverted light microscope (Kern OCL-2) suitable
for observation of live biological samples is integrated into the experimental platform. Digital
camera (FLC 1600H, CMOS pixel size 1.34 m) with a resolution of 16 MP offers for 60x
objective an effective magnification (without digital zoom) at the level of 5800 (Fig.1, part
C2). A typical size of the biological object under observation Saccharomyces cerevisiae cell
with 7 m diameter spans 4 cm (90 pixels; effectively with respect to the resolution limit 18
pixels). For analysis of static microscopic cell images, as well as movies, semi-automatic
image procedures are developed under Matlab and open programming system "Cellprofiler".
Turbidity measurements are realized by a sensor (Fig.1, part D1) with integrated LED light
source and are based on measurement of transmitted light attenuation sensed by a photodiode
detector.
A key part for impedance spectroscopy is a probe. For flow applications, we are developing
probe element that is inserted in between tubing. It consists of plastic body and conductive
electrodes (Fig. 1, part E1). The probe is designed as 3D printing object. The main advantage
is increased flexibility and simplicity during design and testing. 3D printing is continually
improving; currently with accessible spatial resolution at 20 m. 3D printing conductivity
filament materials are gradually approaching conductivity of metals. As an instrument serving
for measurement of impedance spectroscopy portable Stemlab is used. Basically, the device is
a 2-channel oscilloscope and generator based on dual core arm cortex processor, Ethernet
connectivity, and open source software platform. It is used together with LCR meter software
module and impedance analyzer extension board.

4. Discussion and Conclusions

The presented experimental platform will serve for investigation of cells in the liquid phase.
Thus all monitoring equipment is built for bulk samples. However, the flexibility of the
system is required; various elements and sensors may be added or replaced. Static observation

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instead of flow circuit or sequential switching between flow and static phase will be provided.
Later on, it is planned to modify the measurement system from observations of bulk samples
to investigation of planar structures of human cancer cell lines. The aim is to study electrical
processes in cells from the point of view of their diagnostic and therapeutic potential.

Fig. 1. Scheme with elements of the experimental platform arranged along a flow circuit. A cell suspension
together with stirring and magnetic field exposition with Helmholtz coils. B peristaltic pump
providing circuit flow. Optical unit: C1 - inverted light microscope. C2 sample of Saccharomyces
cerevisiae cells under 60x objective. Turbidimetry unit: D1 flow sensor, D2 data acquisition
Arduino-like board, Impedance spectroscopy unit: E1 3D printing flow sensor with built-in
electrodes, E2 - impedance analyzer extension board, E3 - Stemlab board with software impedance
module.

Flexibility also consists in the modification of the exposition part. While keeping the current
source and amplifier untouched, one can obtain DC or AC electric or magnetic field by
replacing coils or adding another suitable EMF applicator. Either separately, or as their
mutual combination, such configuration is suitable for investigation of effects of simultaneous
actions of several components of EMF as proposed in various interaction models.

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The advantage of 3D printing impedance probe is in the efficient testing of different geometry
and scale of the sensor. By miniaturizing of the sensor, we will approach towards electro-
impedance spectroscopy microfluidic chips with dimensions bellow 100 m suitable for
single cell applications [5]. For further construction variants, we plan to integrate 4-wire
impedance measurements together with electrodes suitable for application of a pulsed electric
field. Moreover, multichannel impedance measurement will be utilized in a flow setting in
such a way, that it will be realized before and after application of EMF, also with emphases to
repetitive pulsed EMF.
Besides concentration, the influence of external EMF can also impact certain electrical
properties of cells under investigation, while the respective changes are detectable by
impedance spectroscopy [5]. For these purposes, sensitivity analysis of impedance
measurement is under development, both experimentally and analytically. A spherical model
of cells with varying concentration [6, 7] and varying electrical properties of subcellular
structures like resistivity and capacity of the cell membrane together cytosol resistivity [5] is
involved. To expand information content from impedance approach, later on, nonlinear
dielectric technique will be utilized for yielding impedance response for broader frequency
interval.

Acknowledgements
The study was supported by the research grant 2/0138/16 from the VEGA Grant Agency,
Czech Science Foundation, project no. P102/15-17102S, bilateral exchange project between
Slovak and Czech Academies of Sciences, no. SAV-15-22, and by the "Biomedical Center
Martin" project (ITMS code: 26220220187, project co-financed from EU sources). Authors
also participate in COST Actions BM1309 and CA15211.

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