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Andriole KP, Gould RG, Avrin DE, Bazzill TM, Yin L, Arenson RL.
Continuing quality improvement procedures for a clinical
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radiography-based radiology department. Proc SPIE 1995;
2432:588–599.
See also PHOTOGRAPHY, MEDICAL; RADIOLOGY INFORMATION SYSTEMS;
ULTRASONIC IMAGING.
PIEZOELECTRIC SENSORS
YANBIN LI
Department of Biological and
Agricultural Engineering,
University of Arkansas
Fayetteville, AR
XIAO-LI SU
BioDetection Instruments LLC
Fayetteville, AR
INTRODUCTION
Piezoelectric sensors are generally referred to as analytical
devices for detection of chemical or biological agents with
piezoelectric quartz crystals (PQCs) as transducers. The
origin of piezoelectric sensors can be traced back to 1880
when Jacques and Pierre Curie discovered normal and
converse piezoelectric effects (1). In the former, the appli-
cation of a mechanic stress to the surface of quartz and
some other crystals induces an electric potential across
the crystal; in the latter, conversely, the application of a
voltage across the crystal results in an internal mechanical
360 PIEZOELECTRIC SENSORS
strain. The converse piezoelectric effect is the basis of all
piezoelectric sensors.
PQCs have been widely used in oscillators and filter
circuits for high-precision frequency control. The use of
PQCs as a mass sensor is based on the work of Sauerbrey
(2), which established a linear relationship between the
decrease in resonant frequency and the increase in surface
mass loading of a PQC. Because of its high sensitivity for
mass detection, a piezoelectric sensor is usually called
quartz crystal microbalance (QCM). QCM was originally
and is still used to measure thickness of coatings in vacuum
and air. The first example for application of QCM to
analytical chemistry was reported by King in 1964 (3).
He coated PQCs with various materials and used them
as a sorption detector in gas chromatography to detect and
measure the composition of vapors and gases. The applica-
tions of piezoelectric sensors were limited to the determi-
nation of environmental and other gas species for a long
time. The liquid-phase measurements began in the 1980s
when new oscillator technology emerged and advanced to
make PQCs oscillate in solution as stably as in gas. Then,
numerous piezoelectric chemical sensors and biosensor
have been reported.
Piezoelectric sensors, characterized by their simplicity,
low cost, high mass-detection sensitivity, and versatility,
have found increasing applications in biomedical analyses.
The objective of this article is to briefly present the theory,
equipment, and applications of piezoelectric sensors in the
biomedical area. Interested readers are referred to some
key review articles (4–9) for more theoretical and technical
details of piezoelectric sensors.
THEORY
Typically, a piezoelectric sensor is fabricated by modifying
a PQC with a layer of sensing material, chemical or bio-
logical, that has specific affinity to a target analyte. The
specific binding between the sensing material and the
target analyte causes a change in the resonant frequency
of PQC that is proportional to the amount of target analyte
adsorbed or bound on the sensor surface, which can be
correlated to the concentration of target analyte in the
original sample.
A typical PQC consists of a quartz crystal wafer and two
excitation electrodes plated on opposite sides of the crystal.
The wafer is cut from a natural or synthetic crystal of
quartz. The electromechanical coupling and stresses
resulting from an applied electric field depend on the
crystal symmetry, cut angle, and electrode configuration
(4). Different modes of electromechanical coupling lead to
different types of acoustic waves, including thickness shear
mode (TSM), surface acoustic wave (SAW), shear horizon-
tal (SH) SAW, SH acoustic plate mode (APM), and flexural
plate wave (FPW).
The TSM device, widely referred to as QCM, is the
simplest and most popular piezoelectric sensor. Hence,
we will focus on TSM piezoelectric sensors in this article.
A TSM sensor typically uses AT-cut quartz as a piezo-
electric substrate, which has a minimal temperature coef-
ficient and is obtained by cutting quartz crystals at
approximately 358 from the z-axis of the crystal. Figure 1
AT-cut quartz crystal
Excitation electrodes
Leading wires
Holder
Figure 1. Schematic of an AT-cut piezoelectric quartz crystal.
shows the schematic of an AT-cut PQC. When an alternating
electric field is applied across an AT-cut quartz crystal
through the excitation electrodes, the crystal produces a
shear vibration in the x-axis direction parallel to the
electric field and propagation of a transverse shear
wave through the crystal in the thickness direction. The
resonant frequency of the vibration is determined by the
properties of the crystal and the contacting medium.
Mass Response
Most piezoelectric sensors are used as mass sensors that
are based on the Sauerbrey equation (2):
2F02
DF ¼ pffiffiffiffiffiffiffiffiffiffi
ffi DM (1)
A mq rq
where DF is the frequency change measured (Hz), F0 is the
resonant frequency of the fundamental mode of the crystal
(Hz), A is the area of the gold disk coated onto the crystal
(cm2), rq is the density of the quartz crystal (2.648 gcm3),
m q is the shear modulus of quartz (2.947  10 11
gcm1s2), and DM is the mass change (g). The Sauerbrey
equation is applicable only to a thin (1 mm) and elastic
film coupled to the crystal surface, where the mass loading
can be up to 0.05% of the crystal mass.
According to the Sauerbrey equation, the magnitude of
frequency decrease corresponding to a mass increase is
proportional to F0 2 ; i.e. the higher the fundamental resonant
frequency, the higher the mass sensitivity. Typical AT-cut
PQCs have F0 ¼ 5  30 MHz with a frequency resolution of
0.1 Hz and a mass sensitivity of 0.056  2.04 Hzcm2ng1.
Thinner crystal wafers have higher F0 and thus have higher
mass sensitivity, but they are also more fragile and thus are
more difficult to manufacture and handle. For one of the
most commonly used AT-cut PQCs, F0 ¼ 9 MHz, A ¼ 0.2
cm2, and the detectable mass is 1.09 ng per Hz, which is
approximately 100 times higher than that of an electronic
fine-balance with a sensitivity of 0.1 mg. As AT-cut piezo-
electric sensors can sensitively detect mass change at nano-
gram levels, they are frequently referred to as quartz crystal
microbalances (QCMs) or nanobalances.
However, as a mass sensor, the QCM does not have
selectivity. To make a selective QCM chemical sensor or
biosensor, the QCM must be coated with a film of chemical
or biological recognition material that is selective to a
target analyte.

Viscosity-Density Effect
When a QCM is employed in liquid phase, in addition to
the mass change, it also responds to other factors such as
liquid density and viscosity, surface energy, viscoelasti-
city, roughness, and surface charge density. For a QCM
with only one side in contact with a Newtonian liquid, DF
is linearly proportional to the squared root of the product
of viscosity (hL) and density (rL) of the liquid (10):
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
3=2
DF ¼ F0 rL hL =pmq rq (2)
For a QCM with simultaneous mass and liquid loading,
DF can be expressed as (11)
2F02 pffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
ffi ðDM=A þ rL hL =4pF0 Þ
DF ¼  pffiffiffiffiffiffiffiffiffiffi
mq rq
In equation 3, the first term is equivalent to the
Sauerbrey equation, and the second term is equivalent
to Kanazawa equation. Equation 3 indicates the addi-
tive nature of mass and viscosity-density effects in
changing the resonant frequency. It also shows that it
is impossible to distinguish the mass effect from the
viscosity-density effect when only the resonant frequency
is monitored.
Some piezoelectric sensors are based on the density-
viscosity change rather than on the elastic pure mass
change. For example, a piezoelectric sensor was used to
detect E. coli based on the gelation of Tachypleus amebo-
cyte lysate (TAL) (12), and the detection range is
2.7  104  2.7  108 cellsmL1. Gee et al. (13) used a
piezoelectric senor for measuring microbial polymer
production and growth of an environmental isolate
obtained from river sediment contaminated with petro-
leum hydrocarbons. The increasing amount of produced
polymer corresponded to an increase in the viscosity of the
liquid, which was directly measurable as the fluid con-
tacts the surface of the quartz crystal in the sensor
system. These methods, although they lack specificity,
are advantageous in that coating on the PQC surface is
unnecessary.
Equivalent Circuit Analysis
Equivalent circuit analysis can provide more detailed
information about the surface/interface changes of a
piezoelectric sensor (11,14–17). A PQC can be repre-
sented by a Butterworth–Van Dyke (BVD) model
(Fig. 2), which is composed of a static capacitance (C0)
in parallel with a motional branch containing a motional
inductance (Lm), a motional capacitance (Cm), and a
motional resistance (Rm) in series. Each parameter has
its distinct physical meaning: C0 reflects the dielectric
properties between the electrodes located on opposite
sides of the insulating quartz crystal; Cm represents
the energy stored during oscillation, which corresponds
to the mechanical elasticity of the vibrating body; Lm is
related to the displaced mass; and Rm is the energy
dissipation during oscillation, which is closely related
to viscoelasticity of the deposited films and viscosity-
density of the adjacent liquid.
PIEZOELECTRIC SENSORS 361
Lm
C0 Cm
Rm
Figure 2. Butterworth–Van Dyke model of a piezoelectric quartz
crystal.
The BVD model can be described by the following
(3)
admittance equations:
YðvÞ ¼ GðvÞ þ jBðvÞ (4)
Rm
GðvÞ ¼ (5)
R2m þ ðvLm  1=vCm Þ2
ðvLm  1=vCm Þ
BðvÞ ¼  þ vC0 (6)
R2m þ ðvLm  1=vCm Þ2
where Y is admittance, i.e., the reciprocal of impedance. Y
is a complex quantity, its real part G is conductance, and
its imaginary part B is susceptance. Both G and B are a
function of the scanning frequency f (v ¼ 2pf) and the four
equivalent circuit parameters. These parameters are
determined by physical properties of the quartz crystal,
perturbing mass layer and contacting liquid, and can be
obtained by fitting the measured impedance/admittance
data to the BVD model using the admittance equations.
Figure 3 shows typical admittance spectra of an unper-
turbed 8 MHz AT-cut PQC in air. The fitted results of F0,
Rm, Lm, Cm, and C0 were 7.99 MHz, 9.6 V, 17.9 mH, 22.2
fF, and 8.2 pF (including parasitic capacitance in the test
fixture) for the quartz crystal (18).
High-frequency admittance/impedance analysis has
been extensively used in surface/interface studies. A sim-
pler way to provide insights into the viscoelastic properties
of a bound surface mass is to simultaneously monitor F0
and Rm or F0 and the dissipation factor D using a quartz
crystal analyzer that is much less expensive than the
impedance analyzer. This method has been applied to
study the behavior of adherent cells in response to chemi-
cal, biological, or physical changes in the environment.
For a QCM in contact with liquid, the change of motional
resistance was first derived by Muramatsu et al. (14) as
follows:
DR ¼ ð2pF0 rL hL Þ1=2 A=k2 (7)
where k is the electromechanical coupling factor.
Simultaneous measurements of DF and DR can differ-
entiate an elastic mass effect from the viscosity-induced
effect. DR is a good measure of the viscoelastic change. For
an elastic mass change, DR will be zero and DF will be
linearly proportional to the mass change in accordance
with the Sauerbrey equation. For a QCM with only one
side in contact with a Newtonian liquid, both DF and DR
are linearly proportional to the squared root of the product
362 PIEZOELECTRIC SENSORS
Conductance (S)
Figure 3. Typical conductance and suscept-
ance spectra of an unperturbed 8 MHz AT-cut
PQC in air.
of viscosity and density of the liquid. Therefore, a pure
viscosity-density change will result in a linear DF  DR
plot. In the presence of a viscoelastic change, the DR  DF
plot will lie between the pure viscosity-density effect line
and the elastic mass effect line or even above the former
(8,18).
EQUIPMENT AND EXPERIMENTS
Traditionally, a piezoelectric sensor (resonator) is driven
by a homemade oscillator, and the oscillation frequency is
measured by a frequency counter that is connected to a
recorder or computer for data collection. The Pierce oscil-
lator with a fundamental mode AT-cut resonator is the
most popular oscillator design type and operates with the
piezoelectric sensor as an inductive element. Now such
oscillators are commercially available. One of the suppliers
is International Crystal Manufacturing (Oklahoma City,
OK), which produces a standard (clock) oscillator for gas-
phase QCMs and lever oscillator for liquid-phase QCMs.
Highly sophisticated, automatic, and microprocessor-
controlled piezoelectric detectors or QCM instruments
are commercially available from several manufacturers
(19). The main commercial systems include QCA-917-
and 922 quartz crystal analyzers (Princeton Applied
Research, Oak Ridge, TN), EQCM 400 series electroche-
mical quartz crystal microbalances (CH Instruments, Aus-
tin, TX), EQCN-700 and -900 electrochemical quartz
crystal nanobalances (Elchema, Potsdam, NY), the PZ-
1000 immuno-biosensor system and PZ-105 gas phase
piezoelectric detector (Universal Sensors, Metairie, LA),
RQCM research QCM (Maxtek, Santa Fe Springs, CA), and
Mark series cryogenic and thermally controlled QCMs
(QCM Research, Lake Forest, CA). These systems are
designed to reliably measure mass change up to 100
mg with a resolution of 1 ngcm2. Most of them are
programmed and controlled by easy-to-use Windows-based
software (Microsoft Corporation, Redmond, WA). The QCA
922, designed for EQCM with a potentiostat or stand-alone
0.12 0.06
0.1 0.04
G, measured
G, fitted
0.08 B, measured 0.02
Susceptance (S)
B, fitted
0.06 0
0.04 –0.02
0.02 –0.04
0 –0.06
7.987E+06 7.990E+06 7.993E+06 7.996E+06 7.999E+06
Frequency (Hz)
operation, can simultaneously measure resonant fre-
quency and resistance of QCM. The RQCM can measure
crystal frequency and crystal resistance for up to three
crystals simultaneously. Moreover, high-frequency impe-
dance/admittance analyzers such as E4991A (Agilent
Technologies, Palo Alto, CA) can be used to obtain the
impedance/admittance spectra of the quartz crystal and
to acquire the equivalent circuit parameters by fitting the
impedance/admittance data to the BVD model as described
earlier.
Previously, for liquid-phase applications, the dip-and-
dry approach is made in the measurement of piezoelectric
sensors, in which the resonant frequency of the same
sensor is measured in gas phase before and after the
sample solution is dipped and dried. This approach does
not need a special fixture to mount the crystal; however, it
is unsuitable for automation and has poor reproducibility.
By mounting the crystal to a dip or well cell, like those from
Princeton Applied Research and International Quartz
Manufacturing, and letting only one side of the crystal
exposed to the test solution, it is possible to monitor the
frequency change in solution in real time.
Flow-through QCM biosensors have drawn increasing
attention due to its ease for automation. For example, Su
and Li (20) developed a flow-through QCM immunosensor
system for automatic detection of Salmonella typhimur-
ium. The QCM immunosensor was fabricated by immobi-
lizing anti-Salmonella antibodies on the surface of an 8
MHz AT-cut quartz crystal with Protein A method, and
then installed into a 70 mL flow-through detection cell. The
flow cell was composed of acrylic, with upper and lower
pieces held together by two screws with O-rings. One face
of the sensor was exposed to the 70 mL chamber that was
connected to a peristaltic pump and multiposition switch-
ing valve. The flow cell was designed to reduce the potential
of air bubbles remaining on the crystal after filling from the
dry state and to allow air bubbles in the liquid phase to pass
out without sticking to the crystal. The oscillation fre-
quency of the QCM sensor was monitored in real time
by a Model 400 EQCM system controlled by a laptop PC
 PIEZOELECTRIC SENSORS 363

search from SciFinder Scholar (Chemical Abstracts) with

Figure 4. Components of an automatic QCM immunosensor
system (top) and the flow cell (bottom).
under the Windows environment. Both the pump and the
valve were controlled by a DVSP programmable timer. A
schematic diagram of the whole QCM sensor system is
illustrated in Fig. 4. The operation of this QCM sensor
system was totally automated. As shown in Fig. 5, negative
frequency shifts exist between every two neighboring phos-
phate buffered saline solution (PBS) baselines, which were
attributed to the specific adsorption of target bacteria onto
the biosensor surface.
APPLICATIONS
Piezoelectric sensors, as simple yet powerful tools, have
been extensively employed in detection of chemical and
biological agents as well as in the study of chemical, elec-
trochemical, and biological interfacial processes. An online
the keyword ‘‘quartz crystal microbalance’’ resulted in 4722
references, and 2203 of them are journal articles published
during 2000–2005. These studies were conducted to develop
(1) antibody/antigen-based biosensors (immunosensors) for
detecting biomacromolecules, viruses, cells, as well as small
molecules; (2) DNA/RNA probe-based biosensors (genosen-
sors) for in situ detection of nucleic acid hybridization; (3)
biosensors based on immobilized enzymes, proteins, recep-
tors, lipids, or whole cells; and (4) chemical sensors based on
inorganic or organic films for measurements of organic
vapors, metal ions, and drugs. Also piezoelectric sensors
reported in these studies were used for (1) studies of adsorp-
tion of biomolecules and living cells by bare QCM or QCM
with functionalized surfaces; and (2) QCM/EQCM investi-
gation/analyses of interfacial phenomena and processes,
including self-assembles monolayers (SAMs), films formed
using the layer-by-layer assembly technique, molecularly
imprinted polymers, biopolymer films, micellar systems, ion
transfer at and ion exchange in thin polymer films, doping
reactions of conducting polymers, electrodedeposition of
metals, and dissolution of metal films.
In the following sections, immunosensors and genosen-
sors, which are most relevant and important to biomedi-
cine, are chosen to discuss the applications of piezoelectric
sensors. Some review articles (4–8,21,22) are available for
more comprehensive information about applications of
piezoelectric sensors.
Immunosensors
One important feature of piezoelectric sensors is that
they can be designed as label-free immunosensors. The

20

Time (min)
PBS
0
0 50 100 150 200 250

-20
Frequency shift (Hz)

PBS
5.6 x106
-40
5. 6 x 107
PBS
-60

-80

-100

-120 5. 6 x 108 CFU/mL PBS

Figure 5. Typical output of an automatic QCM immunosensor system for detection of S.

typhimurium.
364 PIEZOELECTRIC SENSORS
immunosensors taking advantage of antibody-antigen
affinity reaction are among the most promising biosensors
due to their high specificity and versatility. Conventional
immunosensors generally involve the formation of a
sandwich immuno-complex consisting of an immobilized
primary antibody, captured target analyte, and labeled
secondary antibody followed by an optical or electrochemi-
cal measurement to detect the label directly or indirectly.
Piezoelectric immunosensors do not need a labeled anti-
body and are thus much simpler and easier in operation
than the sandwich immunosensors.
The first piezoelectric immunosensors is reported by
Shons et al. (23), who modified a quartz crystal with bovine
serum albumin (BSA) and used it to detect anti-BSA anti-
bodies. Since then, numerous piezoelectric immunosensors
have been reported for the detection of various analytes
from small molecules to biological macromolecules, whole
viruses, and cells. In brief, a piezoelectric immunosensor is
fabricated by immobilizing a specific antibody/antigen on
the surface of an AT-cut PQC. When the immunosensing
surface is exposed to a sample solution, a binding reaction
occurs between the immobilized antibody/antigen and its
complementary part (target analyte). The binding event
can be monitored in situ by QCM based on the change of
surface mass loading and/or other properties such as vis-
coelasiticity, and thus, the target species is quantitatively
detected. Figure 6 illustrates the stepwise assembly and
the principle of piezoelectric immunosensor for direct
detection of the binding of target analyte and immobilized
antibody.
Microbial detection is probably the most common area in
which piezoelectric immunosensors are applied. Current
practice for effective treatment of infectious diseases without
abuse of antibiotics relies on rapid identification of specific
pathogens in clinical diagnostics. Nevertheless, conventional
methods for microbial detection are inadequate due to being
tedious and laborious. Although traditional culture methods
hypothetically allow the detection of a single cell, they are
extremely time-consuming, typically requiring at least 24
Au electrode
Surface
modification
Quartz crystal
Sample analysis
Figure 6. Mechanism of a piezoelectric immuno-
sensor for direct detection of the binding of target
analyte and immobilized antibody.
hours and multistep tests to confirm the analysis. Even
current rapid methods such as enzyme-linked immunosor-
bent assay (ELISA) and polymerase chain reaction (PCR) still
take several hours to generate only tentative results and
require skilled personnel.
Numerous piezoelectric immunosensors have been
reported for rapid and specific detection of pathogenic
bacteria as alternatives to the conventional methods since
the pioneer work of Muramatsu et al. (24), which involved
the determination of Candida albicans with an AT-cut
PQC coated with a specific antibody. Piezoelectric immu-
nosensors have been developed for detection of different
bacteria including S. typhimurium, S. paratyphi, E. coli, E.
coli K12, Chlamydia trachomatis, Yersisinia pestis, Can-
dida albicans, and Shigella dysenteriae (25–27). The lower
limits of detection typically ranged between 105 and 107
cells mL1 along with a detection time of tens of minutes to
several hours.
QCM has been used to detect various infectious viruses.
In the study by König and Gratzel (28), Herpes simplex
types 1 and 2, Varicella-zoster virus, Cytomegalovirus, and
Epstein–Barr virus were detected using a reusable QCM
immunosensor with a detection range from 104 to 109 cells.
They reported that a similar QCM immunosensor could
detect Rotavirus and Adenovirus with a linear detection
range from 106 to 1010 cells (25) as well as hepatitis A and B
viruses (29). Kosslinger et al. (30) demonstrated the fea-
sibility of detecting HIV viruses using a QCM sensor.
Antibodies specific to the HIV were absorbed on the crystal
surface, and a serum sample could be detected in 10 min in
a flow QCM system.
A piezoelectric immunosensor was developed for the
detection of cortisol in a range of 36–3628 ppb (31). Cortisol
antibodies were covalently bound onto the Au electrode of
a 10 MHz crystal with a water-insoluble polymer and
thyroxine antibodies.
Piezoelectric immunosensors have also been frequently
reported for determination of biological macromolecules. For
example, Kurosawa et al. (32) constructed a high-affinity
· Silanized layer
· polymer membrane
· Langmuir-Blodgett film
· Protein A
· Self-assembled monolayer
Antibody
immobilization
Immobilized Antibody
Target analyte
Surface mass /
viscoelasticity change
Time

piezoelectric immunosensor using anti-C-reactive protein
(CRP) antibody and its fragments for CRP detection. When
anti-CRP F(ab’)2-IgG antibody was immobilized on the
PQC, the detection limit and the linearity of CRP calibra-
tion curve were achieved at concentrations from 0.001 to
100 mgdL1 even in serum samples.
Antibody immobilization is vital in successful develop-
ment of a piezoelectric immunosensor. The current immo-
bilization methods are mainly based on a silanized layer,
polymer membrane, Langmuir–Blodgett film, Protein A,
and SAM. Protein A, due to its natural affinity toward the
Fc region of IgG molecules, has been commonly used to
orient antibodies for immunoassays. The orient immobili-
zation has the advantage that it does not block the active
sites of the antibodies for binding of target antigens. The
procedure for antibody immobilization based on Protein A
is simple. Briefly, the Au surface of PQC is coated first with
Protein A, and then the antibody is bound to the immobi-
lized Protein A directly. The SAM technique offers one of
the simplest ways to provide a reproducible, ultra-thin, and
well-ordered layer suitable for further modification with
antibodies, which has the potential of improving detection
sensitivity, speed, and reproducibility.
Analytical applications of piezoelectric immunosensors
based on the direct binding between immobilized antibo-
dies/antigens and target analytes are attractive, owing to
the versatility and simplicity of the method. However, the
sensitivity of theses approaches is relatively low due to the
relatively small numbers of analyte entities that can spe-
cifically bind to the limited number of antibody/antigen
sites on the surface. Some amplification techniques have
been investigated for the sensitivity of piezoelectric immu-
nosensors. Ebersole and Ward (33) reported an amplified
mass immunosorbent assay with a QCM for the detection of
adenosine 5’-phosphosulfate (APS). The enzymatic ampli-
fication led to significant enhancement of the detection
sensitivity; levels of approximately 5 ngmL1 (1014 M)
APS reductase could be detected, whereas the direct bind-
ing of APS reductase at even more elevated concentrations
could not be measured. A sensitive QCM immunosensor
was developed by incorporating the Au nanoparticle-
amplified sandwiched immunoassay and silver enhance-
ment reaction (34). Au nanoparticle-promoted silver (I)
reduction and silver metal deposition resulted in about a
two-orders-of-magnitude improvement in human IgG
quantification. Su and Li (18) described a piezoelectric
immunosensor for the detection of S. typhimurium with
simultaneous measurements of the changes in resonant
frequency and motional resistance (DF and DR). In the
direct detection of S. typhimurium, DF and DR were pro-
portional to the cell concentration in the range of 105 to 108
and 106 to 108 cellsmL1, respectively. Using anti-Salmo-
nella magnetic microbeads as a separator/concentrator for
sample pretreatment as well as a marker for signal ampli-
fication, the detection limit was lowered to 102 cellsmL1
based on the DR measurements.
Genosensors
Piezoelectric genosensors are fabricated by immobilizing a
single-stranded (ss) DNA/RNA probe on the PQC surface.
PIEZOELECTRIC SENSORS 365
Au electrode
S
S
S
S
HS
thiolated DNA probe target DNA
Quartz crystal
Figure 7. Illustration of piezoelectric genosensor for in situ
detection of DNA hybridization.
Specific hybridization between the immobilized DNA/RNA
probe and its complementary strand in sample causes a
change in the resonant frequency of the QCM. Various
methods have been used for the immobilization of DNA
probes onto the QCM surface. Among these, the SAM
method is most commonly used because it offers an
ordered, stable, and convenient immobilization. Thiolated
oligonucleotides can directly form a SAM on the gold
surface of the QCM electrode via the Au-thiolate bond.
Figure 7 is an illustration of the piezoelectric genosensor
for DNA hybridization detection.
DNA/RNA probes have been applied to detect patho-
genic microorganisms in clinical samples. Bacterial and
viral pathogens are detectable because of their unique
nucleic acid sequences. The DNA/RNA probing process is
usually preceded by PCR amplification as target nucleic
acid may be present in a sample in very small quantities.
Most PCR formats are followed by the detection of ampli-
cons using gel electrophoresis or the membrane-based
hybridization method. The former lacks of sensitivity;
the latter is more specific, but it requires multistep proces-
sing and is thus time-consuming.
Over the past decade, many attempts have been made to
develop biosensors for sensitive and reliable detection of
DNA hybridization. Fawcett et al. (35) were the first to
develop a piezoelectric DNA sensor. Piezoelectric genosen-
sors, due to their simplicity and cost effectiveness, have
been recently applied to detect gene mutation, genetically
modified organisms, and bacterial pathogens. Su et al. (36)
described a piezoelectric DNA sensor for detection of point
mutation and insertion mutations in DNA. The method
involved the immobilization of ssDNA probes on QCM, the
hybridization of target DNA to form homoduplex or hetero-
duplex DNA, and finally the application of MutS for the
mutation recognition. By measuring the MutS binding
signal, DNA containing a T:G mismatch or unpaired base
was discriminated against perfectly matched DNA at a
target concentration ranging from 1 nM to 5 mM.
The sensitivity of piezoelectric genosensors may be
improved through optimizations of probe immobilization
or by means of signal amplification using anti-dsDNA
antibodies, liposomes, enzymes, or nanoparticles. A nano-
particle is an effective marker for mass amplification as it
has relatively greater mass in comparison with a DNA
molecule. Amplified with nanoparticles, the limit of DNA
detection by QCM can be lowered for several orders to as
low as 1016 M (37). Mao et al. (38) developed a piezo-
electric genosensor for the detection of E. coli O157:H7
with nanoparticle amplification, in which thiolated
ssDNA probes specific to E. coli O157:H7 eaeA gene were
366 PIEZOELECTRIC SENSORS
immobilized onto the QCM surface through self-assembly
and streptavidin conjugated Fe3O4 nanoparticles were used
as ‘‘mass enhancers’’ to amplify the detection signal. As low
as 1012 M synthesized oligonucleotides and 2.67  102
cellsmL1 of E. coli O157:H7 could be detected by the
piezoelectric genosensor.
CONCLUSIONS
Piezoelectric sensors, as sensitive mass sensors or QCMs,
have been applied to detect and measure a broad variety of
biomedical analytes in both gas and liquid phases based on
the adsorption/desorption of target analyte(s) on/from the
sensor surface, in which the selectivity is controlled by the
sensing material. Piezoelectric sensors are more than pure
mass sensors as they are also capable of detecting subtle
changes in the solution–surface interface that can be due to
density-viscosity changes in the solution, viscoelastic
changes in the bound interfacial material, and changes
in the surface free energy. The most attractive advantage
of QCMs is that they are suitable for label-free detection
and flow-through, in real-time detection. However, using
the direct detection approach, the sensitivity of QCM is
inadequate for some applications in biomedical analysis
such as detection of low levels of pathogens and other
biological agents in clinical samples. The lack of sensitivity
may be addressed by employing amplification techniques
as introduced earlier, by using piezoelectric films and bulk
silicon micromaching techniques to manufacture high-
frequency QCMs (21), or by designing devices of other
acoustic wave modes such as SAW, APM, and FPW.
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