Interdigitated array microelectrode capacitive sensor for detection

of paraffinophilic Mycobacteria
Andrew M. Sampson, Erik T. K. Peterson, and Ian Papautsky

Department of Electrical and Computer Engineering

814 Rhodes Hall, University of Cincinnati, Cincinnati, OH 45221

ABSTRACT

Mycobacterium Avium Complex (MAC) is an opportunistic pathogen that threatens public health and has high clinical
relevance. While culture-based and molecular biology techniques for identification are available, these methods are
prone to error and require weeks to perform. There is a critical need for improved portable lab-on-a-chip sensor
technology which will enable accurate and rapid point-of-care detection of these microorganisms. In this work, a new
capacitive sensing strategy is explored utilizing interdigitated array (IDA) microelectrodes and exploiting the
paraffinophilic nature of MAC. In this approach, paraffin wax is deposited over IDA microelectrodes to selectively
extract these microorganisms from samples. As bacteria consume the dielectric paraffin layer, the charging current of
the IDA capacitor changes to facilitate detection. Several IDA geometries were designed and simulated using CFD-
ACE+ modeling software and compared with mathematical models. Capacitance of fabricated devices was determined
using a charge-based capacitance measurement (CBCM) technique. Modeling and experimental results were in good
agreement. Detection of femto-Farad changes in capacitance is possible, making this a feasible technique for sensing
small changes in the paraffin for detection of paraffinophilic MAC.

Keywords: interdigitated array, IDA, capacitive sensor, Mycobacteria, CBCM, CFD-ACE+

1. INTRODUCTION

Interdigitated array (IDA) microelectrodes are becoming increasingly significant in many scientific areas including
biomedical, electrical, chemical, and materials processing, and are proving advantageous over other electrode
configurations. Driven by microelectromechanical systems (MEMS) fabrication technologies, developments have
resulted in a multitude of device applications such as tunable capacitors [1], accelerometers [2], surface acoustic wave
(SAW) transducers [3], comb drives [4], liquid crystal displays [5], strain sensors [6], dielectrophoretic cell separation
[7-8], electroporation [9], chemical sensors [10], batteries [11], label and label-free biosensors and immunoassays [12-
14], DNA detection [15], thermocapillary actuation [16], and material characterization techniques.

Material characterization employing electrodes allows quantification of chemical properties (i.e. electrochemical
activity) and/or electrical properties (i.e. permittivity, conductivity) of target materials through use of voltammetric,
amperometric, impedance or capacitive sensing techniques. IDA microelectrodes are advantageous in electrode-based
material characterization due to the unique geometry they possess. The conventional structure of an IDA electrode
consists of two interleaved, coplanar, comb structures forming a periodic arrangement of adajacent microstrips. A
potential difference applied across each comb structure creates a fringing electric field which extends outside the plane
of the IDA. Physical, chemical, and electrical properties of both the IDA and the surrounding material within the electric
field control the magnitude of faradaic, conductive, and capacitive currents. Passivating the IDA’s boundaries with inert,
electrically insulating materials allows only capacitive current (which accumulates on the IDA’s boundaries) and nearly
eliminates faradaic and conductive currents which only further complicate measurement. Utilizing this arrangement, any
change in current is proportional to a change in the permittivity of the surrounding material within the fringing electric
field. Example applications of IDA capacitive sensors include fluid monitoring [17], resin cure monitoring [18-19],
chemical sensors [20], humidity sensors [21], biosensors [12] and bioparticle detection [22].

Microfluidics, BioMEMS, and Medical Microsystems VI, edited by Wanjun Wang, Claude Vauchier,
Proc. of SPIE Vol. 6886, 68860X, (2008) · 0277-786X/08/$18 · doi: 10.1117/12.768686

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The focus of this work is the development of an IDA microelectrode capacitive biosensor for detection of
Mycobacterium Avium Complex (MAC). MAC is the group of organisms belonging to the genus Mycobacterium and
includes Mycobacterium avium and Mycobacterium intracellulare. MAC is the causative agent of various infections
including gastrointestinal and respiratory tract infections, with children, elderly, and AIDS patients at severe risk. MAC
has been identified in environmental water and hot water systems, soil, animals, hard cheeses, dental devices, and
cigarettes [23]. Current identification procedures for MAC rely on smear tests, culturing, or polymerase chain reaction
(PCR) [24]. Along with lengthy culturing times, the robust outer mycolic acid membrane has shown resistance to certain
staining techniques making this bacteria difficult and time consuming to identify with current detection methods.
Additionally, the bacterium has shown resistance to chlorine and some antibiotics with the emergence of multi-antibiotic
resistant (MAR) strains of Mycobacterium [25]. An improved lab-on-a-chip (LOC) sensing method is essential for a
rapid, accurate, point-of-care identification of these microorganisms. Due to the unique hydrophobic cell envelope
combined with the ability to process long chain aliphatic hydrocarbons, MAC will adhere to paraffin surfaces and are
capable of metabolizing the wax as their sole carbon source when grown in a carbon-free medium. This selective
capture technique, which exploits the organism’s paraffinophilic nature is known as paraffin wax baiting [26]. This
approach has been successfully tested in microfluidic devices and confirmed using fluorescent labels [27]. We describe
a new capacitive sensing strategy exploiting the paraffinophilic nature of MAC. In our approach, a thin film of paraffin
wax is deposited over IDA microelectrodes as a sacrificial baiting layer. As bacteria consume the dielectric paraffin
layer, the IDA microelectrode capacitance values change to facilitate detection.

Methods to measure femtoFarad capacitances are often complicated, require a reference capacitor, or suffer from poor
resolution. Charge-based capacitance measurement (CBCM) was implemented for simplicity and the ability to detect
sub-femtoFarad changes in capacitance [28]. Data extracted using CBCM was compared with simulated values using
CFD-ACE+ and mathematical models developed by Igreja and Dias [29]. Capacitance was determined for nine different
dimensions of two patterns with and without a paraffin wax coverlayer. Comparison of values will allow verification of
an accurate, rapid, new sensing method for detection of MAC and will enable doctors to diagnose infections at point-of-
care for faster and more effective treatment.

2. METHODS
2.1 Fabrication
IDA microelectrodes were patterned and etched in 2200 Å layer of Cr/Au evaporated on a 1 mm thick soda-lime glass
microscope slide substrate (Fisher Scientific) by means of UV photolithography. Two IDA microelectrode patterns were
investigated as shown in Fig. 1 and are referred to as conventional-type IDA microelectrodes and square spiral-type IDA
microelectrodes. Nine IDA microelectrodes of each pattern were fabricated varying only microstrip width, W, and gap,
G. Values of W and G for both patterns ranged from 15 µm to 110 µm. For the conventional-type IDA, constants
include N = 30, L = 3 mm, Wlead = 60 µm, and Gend = 60 µm, where N is the total number of fingers, L is the length of
finger overlap, Wlead is the width of the leads, and Gend is the gap between the tip of the microstrips and the opposing
microstrips' lead. For the square spiral-type, N = 2, D = 15, and Gcenter = G where D is the number of fingers encountered
outward from the center to one side of the IDA (D = 16 on one side), and Gend is the gap at the center. Allowing the
preceding constants establishes W, G, periodicity of microstrips, λIDA, total area, A, and metallization ratio, η, as the only
variables in which λIDA and η are functions of W and G and are given by Equations (1) and (2):

λIDA = 2(W + G ) (1)

W
η= (2)
W +G

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 (a) (b)

(c) (d)
Fig. 1. Two IDA patterns were fabricated: conventional-type (a) and square spiral-type (b). Models were constructed for nine
different dimensions of each pattern and were simulated using Micromesh and CFD-ACE+ electric field solver. CFD
Micromesh models are shown in (c) and (d). Models shown are not to scale and represent a conventional-type IDA with W
= 100 µm and G = 100 µm and a square spiral-type IDA with W = 60 µm and G = 60 µm.

2.2 Simulation
Models were created and meshed in CFD Micromesh (ESI-CFD, Huntsville, AL), Fig. 1, and simulated using CFD-
ACE+ electric solver. The number of x-y cell counts varied for each electrode dimension as did the z-boundary and
z-cell count. The total 3-dimensional cell count for all IDAs was roughly 3.5 million cells. All models were
simulated at +5 V driving potential using CFD-ACE+ electric field solver with an average run time of about 20
minutes. Capacitance values were extracted for all models on a soda lime glass substrate (εr = 7.2 F/m, ρ = 2500
kg/m3) with both air (εr = 1 F/m, ρ = 1.2 kg/m3) above electrodes and a 750 nm paraffin wax (εr = 2.2 F/m, ρ = 898
kg/m3) coverlayer above electrodes as shown in Fig. 2.

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Previously shown, the largest changes in capacitance occur nearer the electrode and saturate at about r = 0.2 [29].
Where r is the ratio of coverlayer thickness, tcoverlayer, to IDA periodicity as shown in Equation 3:

t cov erlayer
r= (3)
λIDA
The paraffin coverlayer was kept well below this saturation value to allow larger changes in capacitance for smaller
changes in the coverlayer height. Models were designed to match fabricated devices with a thermally evaporated
paraffin wax film.

Air

Paraffin / Air

J\IT
ti
+Vss — IDA +Vss

Glass

Fig. 2. Representative crossectional view of IDA microelectrodes indicating material and geometrical design parameters
used for both IDA patterns.

Mathematical models have been developed by Igreja and Dias [29], using conformal transformations and the partial
capacitance method and predict capacitance for the conventional-type IDA electrodes. These models do not predict
capacitance for square spiral-type IDA electrodes. Models were solved for the various dimensions and coverlayers using
Mathematica (v.6, Wolfram Research Inc.) and compared with experimental data and CFD simulations for both air and
paraffin coverlayers.

2.3 Measurement
CBCM was achieved with a dual N-channel and dual P-channel matched MOSFET integrated chip (ALD1103PBL,
Mouser Electronics). The measurement circuit is shown in Fig. 3. Two non-overlapping square wave signals, Vp and
Vn, controlled current flow allowing only one direction of current between the supply and ground (source to drain on the
P-MOSFET and drain to source on the N-MOSFET). During t4 – t3, Vn allows the N-MOSFET to conduct, shorting the
drains and the IDA to ground and discharging the IDA. At this time, Vp is held at the supply voltage level, Vss, to
prevent the supply shorting to ground. Allowing t6 – t1 > t4 – t3 ensures that only one MOSFET is conducting at a time.
The IDA is then allowed to charge during t8 – t7, while the N-MOSFET is held off. DC current injected during t8 – t7
was measured through the P-MOSFET for both the IDA in the circuit, IIDA, and the IDA disconnected, Iref. Capacitance
was extracted using the following relationship in Equation 4 [28]:

C IDA =
(I IDA − I ref )
(4)
(Vss * f )

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 Ct
Ct
t

mA
Capacitor

(a) (b)
Fig. 3. CBCM technique demonstrating the square wave signals (a) applied to each gate of the CBCM circuit (b). Utilizing
two non-overlapping signals at each MOSFET gate enables DC current sampling and extraction of the IDA capacitor
impedance.

CBCM operation was verified and calibrated through comparison of commercial capacitor (0.5 pF, 1pF, and 10pF)
measured values utilizing an HP LCR 4236B capacitance meter at 10 kHz and 100 kHz and with CBCM at various
frequencies between 1 kHz and 130 kHz. 70 kHz gating frequency was employed with CBCM in order to allow good
capacitance resolution while maintaining total DC current within the current sampling device’s (Keithley 2400
Sourcemeter) lowest error range (1 µA ± (0.029% + 300 pA)).

3. RESULTS AND DISCUSSION

Experimental results were compared with simulation results for all IDA microelectrodes and with mathematical model
results for conventional-type IDA patterns. Total IDA capacitance was calculated for IDA microelectrodes in air and is
shown in Fig. 4. CFD-ACE+ simulations and mathematical models matched well with experimental data with less than
10% difference in calculated total capacitance. Total capacitance of IDAs was within calibrated range of l to 10 pF with
standard deviation of about 1 fF for n = 5.

Conventional-type IDA Square Spiral-type IDA •Eopolosoot2

•CFD-ACE+
6
• Fopol 11,1,1
• CFD-ACE+

MothlCo,otooI Modli

a.
C,

liii
C
CC 3

CC

'LI
10-30 15-65 15-105
I
50-30 95-25

Width - Gap (pm)

65 50110 90-70 90-110
1525 1565 15-105 50-30 90-30

Width - Gap (pm)

55-65 55-105 90-20 00110

(a) (b)
Fig. 4. Capacitance extracted with CBCM circuit at 70kHz for (a) conventional-type, and (b) square spiral-type IDA
microelectrodes. Width and gap was varied for nine different microelectrodes of each pattern.

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Deposition of a 750 nm paraffin wax coverlayer demonstrated CBCM’s ability to detect femtoFarad changes in
capacitance as shown in Fig. 5 which shows the change in capacitance due to the coverlayer. Experimental values
agreed better with CFD-ACE+ simulations over mathematical models. Limitations on mesh resolution in the 750 nm
paraffin coverlayer and electrostatic solver solutions are possible reasons for inconsistency with experimental values.
Simulations are more accurate for equal width and gap IDA microelectrodes (η = 0.5) due to x-y plane mesh layout in
Micromesh and are still somewhat accurate outside of this metallization ratio. A graph of capacitance vs. η is shown in
Fig. 6. Mathematical model predictions are most accurate within 0.3 < η < 0.6 but are less consistant outside of this
range. IDA microelectrodes with a small microstrip gap and a large width (η > 0.7) and paraffin coverlayer are the least
consistent mathematical model. Mathematical model errors arise from the fact that the model is only a predicted value
and fails to accurately calculate the electric field distribution as G → 0. This is possibly due to the field extending
farther out of the paraffin coverlayer than the transformation method predicts, accounting for the increase in predicted
capacitance change.

Square Spiral-type IDA

Conventional-type IDA
750 nm paraffin wax coverlayer
750 nm paraffin wax coverlayer • Fepen menta
• Exper me
•c DACEC 0.00
MatEemat ca Made
0.07

0.06

0.05

0.04
0.4

IL]1Ee
0.3 0.03

0.2 0.02

0J I — I I
1030 1565 15105 5030 C525 5565 50110 C070 C0110
0.01

15-25 15-65 15-105 50-30 90-30 55-65 55-105 90-70 90-110

Width - Gap (pm) Width - Gap (pm)

(a) (b)
Fig. 5. Change in capacitance of conventional-type (a) and square spiral-type (b) IDA microelectrodes after depositing a 750
nm film of paraffin wax over the microelectrodes. FemtoFarad changes in capacitance were detectable using the
CBCM technique.

Conventional-type IDA

U
C
c 3
• Experl menta

2 • CFD ACE*

Moheo co Mode

o o.i 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9

n
Fig. 6. Dependence of metallization ratio, η, on total capacitance of a conventional-type IDA.

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 Conventional-type IDA
750 nm paraffin wax coverlayer Square Spiral-type IDA
750 nm paraffin wax coverlayer
• caper menta
• CFD ACES

Mathemat cal Mode

00
0.06

0.05
E
0.04
a
C 0.03

0030 1565 15105 5030 0535 5565 50110 0070 °0110 15-25 15-65 15-105 50-30 90-30 55-65 55-105 90-70 90-110

Width - Gap (pm) width - Gap (pm)

(a) (b)
Fig. 7. Change in capacitance of conventional-type (a) and square-spiral-type (b) IDA microelectrodes normalized to total
area of the IDA microelectrode following deposition of the 750 nm paraffin layer.

Change in capacitance per total area of the IDA microelectrode is shown in Fig. 7. The largest change per area is due to
the smallest λIDA of IDA. This is due to an electric field penetration depth of roughly 30% λIDA [30]. Consequentially,
more of the electric field is contained within the very thin paraffin coverlayer for the small λIDA IDA microelectrodes and
therefore the largest changes are seen by these IDAs due to the paraffin (εr = 2.2) replacing air (εr = 1). Additionally,
conventional-type IDA microelectrodes have greater capacitance change per area than square spiral-type IDA
microelectrodes due to better confinement of the field within the coverlayer for the conventional-type IDAs.
Improvements to simulation accuracy are possible if mesh cell count can be increased. Increasing mesh cell count
allows simulations to converge on one solution. However, experimental error must also be eliminated. Sources of error
include power supply error, parasitic capacitance of IDA leads and surrounding IDA microelectrodes, and variations in
the paraffin deposition layer and metal layer thicknesses. Simulation errors are mainly attributed to inability to properly
mesh at high resolution and large bounding volume due to computer memory limitations. Mathematical errors lie in the
fact that they are only estimated models of capacitance and can only be improved by means of better mathematical
predictions.

4. CONCLUSIONS
Detection of femtoFarad changes in capacitance of an IDA capacitive biosensor for detection of MAC has been verified.
Various IDA patterns and dimensions were investigated to ascertain the best configuration and experimental values
matched well with simulated and mathematical models. To assist in detection of small changes in capacitance arising
from changes of a very small film of paraffin, large electric fields at a short distance are appropriate. Small values of
IDA wavelength, λIDA, allow the field to concentrate within this small film, and have greater effect on the IDA
capacitance. Incorporating these small wavelength IDA microelectrodes into a microchannel enables the possibility for a
high resolution capacitive biosensor for MAC detection. Alternatively, replacement of paraffin (εr = 2.2) with a water
layer (εr = 78), instead of with air (εr = 1) facilitates a larger change in capacitance (>10 pF) as the water replaces
paraffin during MAC baiting and paraffin consumption. A larger change in capacitance eliminates the noise associated
with current sampling at low signal levels. Additionally, eliminating the parasitic capacitance of CBCM cable leads and
the IDA circuit switch will allow more accurate, high signal-to-noise detection. Reducing parasitic capacitances due to
cables will allow clocking speeds above 100 kHz and higher sensitivity. Proper operation also requires a high quality
paraffin wax film to prevent water from contacting the microelectrodes and introduction of faradaic currents. This is
possible with deposition of an insulating layer, but may decrease the sensitivity of the sensor as the sacrificial layer is
moved farther above the plane of the microelectrodes. Immobilization of the target bacteria at high electric field
concentrations (i.e. corners and small localized volumes) will also increase the sensitivity of the sensor.
Dielectrophoresis may need further investigation as it may have beneficial or non-beneficial effects on the MAC

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biosensing strategy. Overall, the most important factors for high sensitivity detection of MAC are the IDA
microelectrode dimensions, such as periodicity, and paraffin coverlayer thickness.

ACKNOWLEDGEMENTS

This work was supported by the National Science Foundation (CBET-0428600) and the University of Cincinnati
Institute for Nanoscale Science and Technology.

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