IMPEDANCE SPECTROSCOPY CELL ANALYSIS IN

MICROCHANNELS
2 1
Shady Gawad\ Stefan Metz\ Laurent Schild & Ph. Renaud
1Swiss Federal Institute of Technology, EPFL DMT-IMS, Switzerland.
2IPHARM, Universite de Lausanne, Switzerland.
shady.gawad@epfl.ch

Abstract
This paper presents a micromachined high throughput flow-cytometer capable of
discriminating cells using their electrical parameters as measured by impedance
spectroscopy. The system is based on a set of electrodes integrated in a microchannel to
measure the differential impedance in two successive segments as the cell passes
consecutively through each. This analyzer is intended to drive a sorting actuator
realising a subsequent cell separation. Size reduction and integration of functions are
essential in achieving precise measurements and high throughput.

Keywords: Cell, impedance spectroscopy, tlow-cytometry, electrodes, microtluidic.

1. Introduction
The device is based on a glass-polyimide mierofluidic chip with integrated channels
and electrodes microfabricated at the length scale of the particles (1-20 )lm) to be
investigated. A laminar liquid flow transports the suspended particles through the
measurement area. Each particle's impedance signal is recorded by a differential pair of
microelectrodes using the cell surrounding
Flow Profile
media as a reference(Fig.l). The integration ::::::>',
of microelectrodes in capillary channels ~.:)
0 _
allows manipulation and detection [1] at a ';c/ ~~
single particle level without the inherent r----/k..,A~-~EI-ect-ro-de-s-~X"l'c------/f,...--
complexity of optical tools. Therefore the ~ \S> ®
device can be considered for numerous
diagnostic and research applications in
oncology, haematology or toxicology;
where, such this cytometry chip would
permit automated and highly controlled Fig. 1 a) Side view of the mierochannel
measurement of biological cell showing a hydrodynamically focussed
characteristics at a much lower cost. particle passing over three electrodes (A,B
However, the use of transparent substrate and C). The impedance signal is measured
and cover materials allow the combination differentially (ZACZSc). b) Impedance
signal. As the distance between the two
of electric impedance and optical detection
measurements areas and time Ttf is known,
techniques (fluorometry).
the speed of the particle can be calculated. *

253
1.M. Ramsey and A. van den Berg (eds.), Micro Total Analysis Systems 2001, 253-255.
© 2001 Kluwer Academic Publishers.
2. Theory
The impedance change I::.Z due to a particle passing through the detection area of the
microchannel is measured and recorded for several excitation frequencies
simultaneously. Placement of the electrodes and concentration of the current lines
around the cell is optimized to achieve a high I::.Z/Z ratio. To fully understand the effects
of the electrode and channel geometry as well as the dispersion due to the position and
size of the measured particle, we have performed 3D FEM electrical simulations of the
detection channel [2]. These simulations were useful in determining the best trade-off
between electrode design and process technology, and for understanding the particle
position and size influence on the detection signal. It showed that the particle position
influence could be compensated using at
least two excitation frequencies and the
measured speed of the particle. +

3. Experimental
The chip was microfabricated on a glass
substrate using polyimide defined
channels and Ti-Pt electrodes (Fig.
2)[3]. The electronic detection circuit
produces a continuous excitation signal
by summing two or more sine waves
(l00kHz-15MHz, -100 mVp-p) at
frequencies of specific interest. This
method allows for a good signal-to-
noise ratio when combined with a
synchronous demodulation technique
(in-phase and quadrature) [4].
4. Results and discussion
a)
o
Fig. 2 a) The microfabricated chip with a
PDMS cover and molded fluidic connections.
b) Chip-on-chip configuration designed with
two outlets, top and bottom electrodes and an
experimental sorting chamber. *
5 & 8 um latex beads impedance
1.8
at a frequency ot 1.72 MHz
1.6

1.4
To characterize the device and verify
that it could in effect differentiate cell I 1.2 Sum

or particle subpopulations in a mixed ~

Bum

_.. .. . ',(- . . . . .
c 0.8
sample, we used calibrated latex beads ~ . ...
_.~
of 5 and 8 ~m (Serva) and human 0.6
....
.. ..- ._.. --
erythrocytes as well as erythrocyte 0.4

ghost cells (Le. cytoplasm replaced by 0.2

the PBS solution). The chip also 00 0,01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09 0.1
Signal amplitude IV]
provides a measure of each particle
speed, which is very useful in Fig. 3 Transit time vs. measured amplitude
determining the particle position in the signal of 5 and 8 ~m latex beads (2000
channel. Plotting of the particle transit recorded events). The particles were mixed in
time against the measured signal a standard PBS solution and passed through
amplitude shows that the average the measurement channel. *

254