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phase gradients to the electric field. The Clausius- For a positive value of Re[K(o)], the particle is
Mossotti factor is defined as attracted to high electric field gradients with pos-
itive dielectrophoresis (pDEP). With dielectro-
ep em phoresis it is possible to manipulate particles
K o (2)
ep 2em using a variety of techniques and applications.
polarizes, the dipole moment will align the parti- previously mentioned dielectrophoretic manipu-
cle with its longest nondispersed dipole parallel lation techniques can be applied to determine
to the field lines. Its orientation is a function of particle dielectric characteristics. By varying
the electric field frequency and the dielectric both the medium conductivity and the applied
properties of the medium and particle. frequency, the changes in a particles induced
Dielectrophoretic forces, though, can be used dielectrophoretic behavior can be visually
to not only rotate a particle, but trap it as well. observed.
There are two types of particle trapping, those The electric field can induce higher order
that utilize pDEP or nDEP forces. For example, poles, called multipoles, instead of the assumed
four electrodes can be positioned in a quadrupole dipole. This occurs when the electric field is
arrangement and, when the appropriate electric highly nonuniform, for example, when the elec-
field is applied, a particle or particles are trapped trode geometry is on the same length scale as the
in the electric field null at its center (Fig. 2c). particle or when a particle is in a field null. For
Particle trapping will be revisited later in this most applications, though, the dipole assumption
entry. Traveling wave dielectrophoresis is the accurately depicts the behavior of the particle.
linear application of electrorotation (Fig. 2d). However, multipoles can effect the particles
An AC electric wave is produced by applying interaction with the electric field and will react
an electric field that travels linearly along differently from an induced dipole. The electric
a series of electrodes. The particle will translate field can also induce some negative physiological
in the same or opposite direction as the traveling effects to the cell by inducing cell heating and
wave depending on the properties of the applied influencing transmembrane voltage. Transmem-
signal frequency and the dielectrics of the particle brane voltage can affect ion movement and dam-
and medium. age voltage-sensitive proteins. The electric field
itself can heat the surrounding medium in what is
Modeling and Dielectrophoretic Effects called Joule heating. Joule heating creates tem-
When incorporating dielectrophoresis in Lab-on- perature gradients that can directly heat the cell or
a-Chip systems, additional modeling parameters create localized convection currents. By working
and electro-physiological interactions need to be at the micrometer length scale, some of these
considered. Obviously, biological cells and some negative effects are minimized; however, they
particulates are not all completely spherical nor may still influence the system and cannot be
are they homogeneous. Typically cells are ignored.
modeled as multi-shelled particles with each
shell having its own respective conductivity and
permittivity. Additionally, nonspherical particles Key Research Findings
are modeled as ellipsoids. For each of these situ-
ations, the Clausius-Mossotti factor is extended This dielectrophoresis background only serves as
to include these geometrical and layered effects a brief overview; these technologies need to be
by applying polarization factors. These modifica- explored in more detail before incorporating
tions are described in detail elsewhere [24]. The them into a Lab-on-a-Chip system. Lab-on-a-
dielectric properties of some particles and cells Chip systems integrate techniques of small fluid
are unknown; however, dielectric techniques can and sample handling with detection or process
be applied to determine these characteristics. For capabilities. Dielectrophoresis can be incorpo-
example, when an induced particle changes from rated into these systems to manipulate, separate,
pDEP to nDEP, or vice versa, this is called its or trap cells as well as control small amounts of
crossover frequency. Similarly, a particle in fluid. This technology can be used to trap cells for
electrorotation that changes in rotational velocity additional analysis, separate cell types based on
or direction under different conditions will give dielectric properties, dispense picoliter droplets,
insight into its dielectric properties. Many of the or used for similar manipulative applications.
AC Dielectrophoresis Lab-on-Chip Devices 5 A
Electric Potential
dielectrophoretic trapping force can be estimated
with the hydrodynamic drag necessary to release
the particle. Similarly, interdigitated electrodes A
n DEP p DEP
can be used to separate two cells that experience
particle particle
opposite pDEP/nDEP forces due to their different
V V dielectric characteristics. Interdigitated electrodes
are one of the simpler electrode geometries, com-
AC Dielectrophoresis Lab-on-Chip Devices, pared to other trapping arrangements.
Fig. 3 Dielectrophoresis of particles with interdigitated
Other electrode geometries have been
electrodes (side view). Particles experiencing pDEP are
attracted to the edges of the electrode, while nDEP forces designed to capture individual cells using either
repel the particle to the middle of the electrode pDEP or nDEP forces. A number of these geom-
etries have been explored by Voldman and his
research group [5] and will be discussed here. For
However, dielectrophoresis itself cannot be used pDEP, geometries are created that generate
as a sensor, except to determine dielectric prop- extreme electric field gradients in the trapping
erties of cells or detect such changes in response region. A geometry that creates such a field gra-
to stimuli. The integration of sensory technolo- dient is illustrated in Fig. 4a. An electrode smaller
gies with dielectrophoresis is not discussed. than the diameter of the particle has the signal
Instead, specific novel dielectrophoresis applica- applied through it to a larger, receiving electrode.
tions are addressed in detail. The generated electric field would be similar to
that illustrated in Fig. 1, with a high gradient in
Trapping Particles proximity to the smaller electrode. Electric field
One of the more popular dielectrophoretic manip- nulls have also been generated with electrode
ulation techniques involves the trapping of indi- geometries for nDEP traps. Recall the quadrupole
vidual or groups of cells. Dielectrophoresis can be geometry in Fig. 2c. When an appropriate electric
thought of as electrical tweezers that will grab field is applied, a field null is created in the center
onto and position cells. Trapping can be accom- of the trap. The trapped particle is repelled from
plished by means of novel electrode geometries the surrounding regions of field gradients.
that utilize pDEP or nDEP forces to selectively Another nDEP trap is illustrated in Fig. 4b. The
capture a particle or cell. Some electrode geome- electric field is applied from the trapping elec-
tries induce both types of forces simultaneously; trode to the receiving electrode strip. A particle is
an example of such an arrangement involves inter- trapped within the generated null inside the
digitated electrodes (Fig. 3). Planar electrode fin- boxed electrode geometry. These previous
gers, whose width and gap separation are a few geometries will trap a particle near the surface
times larger than the diameter of the particles of of the substrate. Electrodes, though, can be
interest, have alternating applied voltages (either arranged three dimensionally to create traps that
180 out of phase or alternating + V/ground). This capture particles and cells in suspension. One
geometry creates a strong pDEP force at the edges example is a nDEP trap using two stacked quad-
of the electrodes and a field null above the center rupole geometries, resulting in an octopole trap
of each electrode strip. This arrangement provides (Fig. 4c). The electric field null is generated in the
both trapping capabilities and the ability to deter- midplane between these two electrode substrates,
mine unknown particle dielectric properties from and thus, a particle experiencing nDEP is cap-
visual observation. For example, at a particular tured in suspension. These are just a few exam-
frequency and medium conductivity, a particle ples of established trapping geometries, as others
will experience a pDEP force. However, for could be designed to suit a particular trapping
a different applied frequency and/or a change in application.
medium conductivity, the same particle could There are numerous trap geometries that exist
undergo nDEP. The strength of the for both pDEP and nDEP. These electrode
A 6 AC Dielectrophoresis Lab-on-Chip Devices
AC Dielectrophoresis
Lab-on-Chip Devices,
Fig. 4 Various electrode
geometries for
dielectrophoretic traps:
(a) pDEN trap (side view),
(b) nDEP planar trap
(top view), and (c) nDEP
octopolar trap
let
Electrodes
ut
varied to trap both single cells and groups of
O
cells alike. However, there are dielectrophoretic
interactions that cannot be ignored for trapping
Outlet
Inlet
applications. Multipoles may need to be consid-
ered for single-particle traps since the generated
field will have inhomogeneities on the scale of
Flow Direction
the particle itself. Also, particle-particle
dielectrophoretic interactions will influence the
AC Dielectrophoresis Lab-on-Chip Devices,
capabilities of multi-particulate traps. Dielectro-
Fig. 5 An illustration of a dielectrophoretic microsystem
phoresis, therefore, needs to be accurately that can selectively sort particles (top view). Recall that
modeled and thoroughly investigated prior to the electrodes are on the top and bottom surfaces of the
use in a specific application. microchannel and to not mechanically manipulate the
particles
Electrode-Based Systems
Microsystems have incorporated dielectro- selective biological investigations. An example
phoretic forces into microchannels to manipulate of a dielectrophoresis system and a description of
particles and cells with various electrode geome- its potential for biological analysis are explained
tries. Electrodes can be used to align target cells in [6]. These dielectrophoresis techniques can be
or separate particulate cells from the rest of the coupled with a variety of existing biological tools
sample, an example of which is illustrated in for innovative applications including fluores-
Fig. 5. These systems use a three-dimensional cence evaluation of cells in small populations,
arrangement of electrodes, with mirrored elec- cell sorting, long-term investigations of single
trode geometries aligned and separated by cells, cellular kinetics, and other similar
microfluidic structures. Thus the electrodes in Lab-on-a-Chip analyses.
Fig. 5 are on the top and bottom surfaces of the
microchannel and do not significantly impede or Insulating Dielectrophoresis
manipulate the fluid flow. This system is opti- Conventionally, the nonuniform electric fields
mized for a particular set of cells such that the for dielectrophoresis applications are produced
applied signals generate the strongest dielectro- directly with an arrangement of conductive elec-
phoretic forces; this system will operate with the trodes. However, a nonuniform electric field can
maximum possible applied flow rate for more be generated in microchannels without direct
effective processing. The system, though, will interaction with electrodes. For example, Cum-
be selective to a set of cells with the same or mings generated nonuniform electric fields using
similar dielectric properties. This process is obvi- remote electrodes placed at the inlet and outlet of
ously advantageous for an application that selec- the microchannel [7]. The electric field is applied
tively separates cells based on their dielectric across the channel and is deformed by the varia-
properties. Additionally, this system can simulta- tions in the channel geometry or by insulating
neously separate and trap targeted cells for structures or posts within the microchannel.
AC Dielectrophoresis Lab-on-Chip Devices 7 A
These mechanical structures within the channel Illumination
will affect the generated electric field as well as Source
dynamically change the flow field; these can be A
successfully coupled to produce an efficient par-
ticle concentrator within the channel. By using
remote electrodes, electrochemical reactions and Glass/ITO
S
other unwanted dielectrophoretic effects will not Photoconducter +++ i
occur within the section of particle collection or g
concentration. However, this technique relies on n
the application of higher voltages to produce the Channel a
necessary electric field gradient, with magnitudes l
Glass/ITO
in the hundreds to thousands of volts (outside the
range of conventional waveform generators).
AC Dielectrophoresis Lab-on-Chip Devices,
The driving mechanism for fluid flow for these Fig. 6 An example for a simple setup for optically
systems is electrokinetic in that it combines fea- induced dielectrophoresis. The optical source is illumi-
tures of electroosmotic and electrophoretic flows. nated through the ITO/glass substrate onto the photocon-
Insulating dielectrophoresis Lab-on-a-Chip sys- ductive layer, creating an electric field gradient in which
particles experiencing pDEP are attracted
tems use a coupled arrangement of electrokinetic
and dielectrophoretic phenomena to separate or
capture targeted particles. Consider such the photoconductive layers have higher conduc-
a dielectrophoresis system that has an array of tivities under illumination, thus distorting the
insulating posts within a microchannel. At a low otherwise uniform electric field. An illustration
applied electric field, there will be induced elec- of a simplified optically induced dielectro-
trokinetic motion but minimal particle manipula- phoresis system is shown in Fig. 6. The illumina-
tion effects from dielectrophoresis. However, at tion of the photoconductive layer produces
higher magnitudes the influence of insulating a virtual electrode and locally induced dielectro-
posts becomes apparent, inducing dielectro- phoretic forces. These forces can be used in
phoretic forces on the particles. At a certain pDEP or nDEP applications and have the distinct
threshold in the applied electric field, dielectro- advantage of being temporary, as opposed to
phoresis forces on the particles are greater than permanent microfabricated electrode features.
the electrokinetic forces, resulting in captured For example, trapped particles can be moved
targeted particles at the insulating posts. Releas- with the translation of the illumination pattern
ing these particles is just a simple decrease in and released by simply turning off the light.
signal magnitude. Structures other than posts These optical methods, though, have been
have been successfully used to similarly manip- developed to include image projectors to manip-
ulate and separate particles of different dielectric ulate cells using larger, patterned illumination
properties. areas. For example, virtual structures can be
used to direct cells or can be alternatively used
Optical Dielectrophoresis to pattern trapped cells. An investigation of opti-
An electrode-less approach to induce localized cally induced dielectrophoresis for cellular
dielectrophoretic forces can be created by light manipulation is found in detail elsewhere
illumination in optically induced dielectro- [8]. This investigation characterizes the control-
phoresis. This technique utilizes low-power opti- lability of these optical techniques. This tech-
cal beams to manipulate particles and cells. nique, though, should not be confused with
Consider an AC electric signal applied across optical tweezers (also called optical traps),
two parallel electrodes with a film of photocon- which manipulate cells with forces generated by
ductive layer in between. This generates a highly focused laser beam. Optical tweezers use
a uniform electric field across the film; however, high optical intensity that may cause damage to
A 8 AC Dielectrophoresis Lab-on-Chip Devices
Electrophoresis Definition
Electrothermal Effects
Joule Heating in Electrokinetic Flow: AC electroosmosis (ACEO) is a nonlinear elec-
Theoretical Models trokinetic phenomenon of induced-charge elec-
Lab-on-a-Chip Devices for Particle and Cell troosmotic flow around electrodes applying an
Separation alternating voltage.
Lab-on-a-Chip (General Philosophy)
Overview
References
Classical electrokinetic phenomena, such as elec-
1. Pohl HA (1978) Dielectrophoresis. Cambridge Uni- troosmotic flow and electrophoresis, are linear in
versity Press, Cambridge
2. Jones TB (1995) Electromechanics of particles. Cam-
the applied voltage and thus cannot produce any
bridge University Press, Cambridge net flow under alternating current
3. Morgan H, Green N (2003) AC electrokinetics: col- (AC) conditions. A variety of nonlinear electro-
loids and nanoparticles. Research Studies Press, kinetic phenomena, which persist in AC fields,
Baldock
have been known for decades in colloid science,
4. Hughes MP (2003) Nanoelectromechanics in engi-
neering and biology. CRC Press, Boca Raton but the focus has been on electrophoretic mobil-
5. Rosenthal A, Taff BM, Voldman J (2006) Quantita- ity and particle interactions. The advent of
tive modeling of dielectrophoretic traps. Lab Chip microfluidics has stimulated interest in the use
6:508515
of electric fields to drive fluid flows, without
6. Muller T, Pfennig A, Klein P, Gradl G, Jager M,
Schnelle T (2003) The potential of dielectrophoresis any moving parts. In this context, nonlinear elec-
for single-cell experiments. IEEE Eng Med Biol Mag trokinetics offers some unique advantages, such
22:5161 as the reduction of unwanted electrochemical
7. Cummings EB (2003) Streaming dielectrophoresis
for continuous-flow microfluidic devices. IEEE Eng
reactions (using AC voltages) and the ability to
Med Biol Mag 22:7584 drive fast, programmable flows at low voltages
8. Chiou PY, Ohta AT, Wu MC (2005) Massively par- (using closely spaced microelectrodes).
allel manipulation of single cells and microparticles In the late 1990s, Ramos et al. discovered
using optical images. Nature 436:370372
steady electroosmotic flow over a pair of micro-
9. Jones TB (2001) Liquid dielectrophoresis on the
microscale. J Electrost 5152:290299 electrodes applying an AC voltage and dubbed
10. Washizu M, Kurosawa O (1990) Electrostatic manip- the effect AC electroosmosis [1]. Around the
ulation of DNA in microfabricated structures. IEEE same time, Ajdari predicted ACEO flow over
Trans Ind Appl 26:11651172
periodic electrode arrays and showed how the
effect could be used for long-range pumping
[2]. As the performance of ACEO pumps has
advanced [3, 4], ACEO has also been exploited,
AC Electro-Osmotic Flow in conjunction with dielectrophoresis (DEP), in
different geometries to manipulate particles and
Martin Z. Bazant cells in microfluidic devices [57].
Department of Mathematics and Institute of
Soldier Nanotechnologies, Massachusetts
Institute of Technology, Cambridge, MA, USA Basic Methodology
A
Current tubes, width D z
Double layer
capacitance
d
Glass substrate Dz
Electrodes Distance, z
AC Electro-Osmotic Flow, Fig. 1 Equivalent RC circuit resistance (due to shorter current tubes) and thus charge
model for double-layer charging over a pair of electrodes. more quickly than the outer edges (Reproduced from [1])
The inner edges of the electrodes encounter less bulk
The main difference with other examples of charges would both change sign, resulting in the
ICEO, such as flows around metal colloids, is same ICEO flow.
that ACEO involves electrode surfaces, which Under AC forcing, the flow peaks when the
supply both the electric field and the induced oscillation period is comparable to the charging
screening charge, in different regions at different time (Fig. 2b). ACEO flow decays at higher fre-
times. For this reason, ACEO is inherently time- quencies, since there is not enough time for
dependent (as the name implies) and tied to the charge relaxation (Fig. 2a). It also decays at
dynamics of diffuse charge, as ions move to lower frequencies, since there is enough time to
screen the electrodes. completely screen the bulk electric field (Fig. 2c).
Perhaps the easiest way to understand ACEO Mathematical models of ACEO follow other
is to consider a pair of planar electrodes applying examples of ICEO, as described in the article on
a sudden DC voltage (which is analogous to nonlinear electrokinetic phenomena. A major
ICEO flow around a polarizable particle in simplification in the case of small voltages is to
a sudden electric field). As shown in Fig. 1, assume sinusoidal response to sinusoidal AC
charge relaxation can initially be described by forcing and solve only for the complex ampli-
an equivalent RC circuit, where the diffuse layers tudes of the potential and velocity components
act as capacitors, connected to current-tube at a single frequency o (Fourier mode) [2]. In this
resistors of varying length through the bulk solu- regime, the basic scaling of time-averaged ACEO
tion. Since the resistance is smaller (and the field flow is
larger) near the gap, the inner portions of double
layers on the electrodes charge more quickly than eV 2
the outer portions. As shown in Fig. 2, this causes <u> / h i2 (1)
ICEO flow to occur, directed outward from the 1 dL ooc ooc
gap, only when the electrodes are partially
screened, as the tangential field from the where V is the applied voltage, e and Z are the
unscreened outer portions acts on induced charge permittivity and viscosity of the liquid (both
on the inner portions. Note that the flow is inde- assumed constant), L is electrode spacing
pendent of the sign of the applied voltage: If the (roughly center to center), and d is the ratio of
polarity were reversed, then the field and induced the diffuse-layer to compact-layer capacitances
A 12 AC Electro-Osmotic Flow
AC Electro-Osmotic Flow, Fig. 2 The basic mechanism where the initial normal current is strongest and causes
of AC electroosmosis: electrochemical relaxation (top) the unscreened field lines to dip down and provide tangen-
and induced-charge electroosmotic flow (bottom) in tial components over the induced charge; the result is
response to a suddenly applied voltage across an electrode ICEO flow directed away from the electrode gap. (c)
pair. (a) At first the electric field has no tangential com- After the charging time passes, the electrodes are fully
ponent on the electrodes, since they are equipotential screened, leaving no electric field and thus no flow. An AC
surfaces, and thus there is no electroosmotic flow. (b) voltage can drive a steady time-averaged flow, similar to
Capacitive double-layer charging begins near the gap (b), if its period is comparable to the charging time
(both assumed constant). The peak frequency is [2, 8]. Although reactions surely play a role in
at the scale of the RC charging time ACEO, high-frequency flow reversal may be
attributable to other effects, such as ion crowding
D1 d in the diffuse layer [9].
oc / (2)
lL
Fluid Pumping
where l is the Debye screening length and D is Regardless of the details of flow generation, there
a characteristic ionic diffusivity. are some useful general principles to guide the
The relevant case of a large applied voltage, development of microfluidic pumps. ACEO flow
V kT/e = 25 mV, is not yet fully understood. over a small set of electrodes can be used for local
A variety of new effects could become important, fluid mixing or particle trapping at stagnation
such as steric effects of finite molecular sizes, points, but the flow decays quickly away from the
viscoelectric effects, discrete electrostatic corre- electrode surfaces. A symmetric, periodic array of
lations, dissociation and de-solvation kinetics, many interdigitated electrodes (of alternating
and/or Faradaic electrochemical reactions pass- polarity at each moment in time) similarly pro-
ing current through the surface. In particular, duces an array of counter-rotating convection
Faradaic charging (as opposed to capacitive rolls, but no net pumping of the fluid in one direc-
charging, described above) has been proposed tion. Instead, long-range pumping over an elec-
to explain experimentally observed flow reversal trode array requires broken symmetry within each
at high voltage [7], although this has not yet been spatial period to rectify the AC forcing.
borne out in a complete mathematical model of There are several ways to design ACEO
ACEO. Standard models of Faradaic reactions pumps by breaking symmetry in a periodic elec-
mainly predict suppression of the flow trode array. Ajdari originally suggested modulat-
(by short-circuiting of the double-layer charging ing either the electrode capacitance via
process) and flow reversal only in certain cases, a dielectric coating (Fig. 3a) or the surface height
at lower frequencies than in the experiments (Fig. 3b) with half the spatial period of the array,
AC Electro-Osmotic Flow 13 A
a FLUID b FLUID
x x
y y
A
AC Electro-Osmotic Flow, Fig. 3 Sketches of local (a) nonuniform surface coatings; (b) nonuniform surface
broken symmetries in a periodic electrode array which height (Reproduced from [2])
lead to global time-averaged ACEO pumping:
so that one side of each electrode drives stronger voltage. At low frequency, a similar induced-
ACEO flow compared to the other side and charge electroosmotic mechanism, which peaks
thus wins to produce net pumping over the at the RC frequency (2), is responsible for the
array [2]. In the first implementation of an flow [12]. At high frequency (or with a thick
ACEO pump, Brown, Smith, and Rennie opted dielectric coating on the electrodes), the classical
instead to break symmetry by using planar elec- Ehrlich-Melcher effect used to pump dielectric
trodes of different widths and gaps [10], and, until liquids, which peaks at the Debye frequency,
recently, this design was the only one studied s/e = D/l2, can also be observed [13]. Although
experimentally [3] or theoretically [8]. traveling-wave ACEO seems to produce slower
The performance of ACEO pumps can be flow than standing-wave ACEO with planar elec-
greatly enhanced by designing appropriate trodes, the possibility of designing suitable
non-planar electrode geometries. As recently non-planar electrodes has not yet been
predicted by Bazant and Ben [11], various 3D considered.
ACEO designs exhibit dramatically increased
flow rate without flow reversal, due to a special
geometry in which the nonuniform slip profile on Key Research Findings
the electrodes all contributes to flow in the same
direction. The basic idea is to create a fluid con- The groups of Ramos [1] and Green [15] have
veyor belt with electrodes, each having steps of pioneered the study of ACEO by developing the-
two different heights: On each electrode, the oretical models and measuring flows around pairs
region of desired forward flow is raised up, of microelectrodes in dilute KCl. Their work has
while the region of reverse flow is recessed ruled out electrothermal effects (below 5 V in
below, so as to recirculate in a vortex aiding the dilute electrolytes) and clarified the capacitive
forward flow (rather than fighting it, as in planar charging mechanism described above. The basic
designs). This can be accomplished with elec- model successfully predicts the scaling of ACEO
trodes having electroplated metal steps, as flow at low voltage and low salt concentration,
shown in Fig. 4, although other designs are pos- although it tends to overestimate velocities.
sible, such as flat electrode steps deposited on Following Ajdari [2], there has been extensive
a grooved surface (without the vertical metal experimental [3, 4, 10, 12, 13] and theoretical
surfaces). Simulations predict that 3D ACEO [8, 9, 11] work on ACEO flows driven by periodic
pumps are faster than planar pumps by more interdigitated electrode arrays. Velocities
than an order of magnitude, at the same voltage exceeding 100 mm/s have been demonstrated in
and minimum feature size, and thus can achieve water [4, 10] and dilute KCl [3] by applying
mm/s velocities with only a few volts. This sug- several volts at 1100 kHz frequencies using
gests using 3D ACEO pumps to drive flows in asymmetric arrays of planar electrodes with
battery-powered, portable, or implantable unequal widths and gaps, at the scale of several
microfluidic devices. microns. Somewhat slower flows have also been
Fluid pumping over electrode arrays can also achieved by applying traveling-wave voltages
be achieved by applying a traveling wave of with interdigitated planar electrodes [12, 13],
A 14 AC Electro-Osmotic Flow
AC Electro-Osmotic Flow, Fig. 4 Top: Simulations of design with stepped electrodes having a symmetric foot-
ACEO microfluidic pumps, showing the time-averaged print and the same minimum feature size; the reverse slip
flow over a pair of microelectrodes (dark regions) in one now recirculates in a vortex to create a fluid conveyor belt
spatial period of an interdigitated electrode array. (a) for the raised pumping flow from left to right (reproduced
A nearly optimal planar design with different electrode from [11]). Bottom: Scanning electron microscopy
sizes and gaps; the smaller electrode has the largest local images of each design fabricated in gold on glass with
slip velocity, but the larger electrode wins in overall minimum feature size (gap) of 5 mm (Unpublished, cour-
pumping from left to right. (b) A much faster 3D ACEO tesy of J. P. Urbanski using the methods of [4, 14])
although the electric connections are more a universal decay with concentration like
complicated. u / log(cmax/c) at large voltage [9]. As shown
A puzzling feature of ACEO in many situa- in Fig. 5b, the flow effectively vanishes above
tions with planar electrodes is that the flow direc- a bulk salt concentration of only cmax 10 mM,
tion can reverse (compared to the theory) with which is an order of magnitude smaller than most
increasing voltage, typically above 2 V. For biological solutions.
example, high-voltage flow reversal has been Nevertheless, advances have been made in
observed in dilute KCl for AC forcing [3] and applying ACEO flows to pumping, mixing, and
traveling-wave forcing [13] and in tap water for trapping particles in microfluidic devices. Exper-
AC forcing with a DC offset [7]. As shown in iments on 3D ACEO pumps with non-planar
Fig. 5a, the reverse flow is eventually faster than stepped electrode arrays have demonstrated an
the forward flow and seems to grow more quickly order of magnitude increase in flow rate versus
with voltage than the expected scaling u / V2. the fastest planar design, due to the robust fluid-
Another puzzling feature of ACEO is the conveyor-belt mechanism [4]. For a wide range
strong decay of the flow with increasing salt of near-optimal step heights, there is also no
concentration. For this reason, all experiments observed flow reversal [14]. Therefore, it is now
in the literature have used either very dilute possible to drive fast, reliable mm/s flows in
solutions (mostly KCl) or water (deionized or water or dilute electrolytes with less than 3 V,
from the tap). A few groups have studied the which can be provided by a small lithium battery
concentration dependence of ACEO in aqueous (Fig. 6).
KCl solutions [3, 15, 16]. These experiments Progress has also been made in exploiting
and recent work on electrokinetic motion of ACEO flows to manipulate colloidal particles
heterogeneous particles suggest that flows and biological cells in microfluidic devices
due to induced-charge electroosmosis exhibit [57]. The basic strategy is to use ACEO flow
AC Electro-Osmotic Flow 15 A
a 6
Electrode b 100
5 damaging 0.001 mM
zone 0.003 mM
A
80
0.01 mM
Voltage (Vrms)
4
0.03 mM
Velocity [m/s]
60 0.10 mM
3 0.30 mM
40 1.0 mM
2
20
1 0
0 20
103 104 105 106 100 101 102
Frequency/Hz Frequency [kHz]
AC Electro-Osmotic Flow, Fig. 5 Experimental data for damaging refers to bubble formation and inability to
ACEO pumping of KCl around a microfluidic loop, observe consistent flows. (b) Velocity versus AC fre-
one-fifth covered by the asymmetric planar electrode quency at 2 V peak to peak for different concentrations
array of Fig. 4a. (a) Contour plot of mean velocity versus (Reproduced from [16]). Both show high-voltage, high-
AC frequency and RMS voltage at a bulk salt concentra- frequency flow reversal, and the latter shows the strong
tion of 0.1 mM (Reproduced from [3]), where electrode decay with salt concentration
Umax [m/s]
1V Planer 8
100
200
50
100
0 0
100 50
1 10 100 0 2 4 6 8 10
Frequency [kHz] h [m]
AC Electro-Osmotic Flow, Fig. 6 (a) Comparison of 5 mm minimum feature size (reproduced from [4]). (b)
ACEO pumping of water around a microfluidic loop by Experimental optimization of the step height for 3D
planar and (non-optimal) 3D electrode arrays with similar ACEO pumping of dilute KCl (Reproduced from [15])
to draw particles to stagnation points on the elec- a DC bias voltage to the low-frequency AC
trodes, where they are trapped, presumably by voltage (50100 Hz) between adjacent electrodes
DEP (although the classical theory does not [7]. Particles are observed to collect only on the
seem to predict this effect). By increasing the positively biased electrode, as shown in Fig. 7b.
voltage, the ACEO flow can be reversed, and It has been suggested that opposing ACEO
particles are observed to move away from the flows are produced by the competition between
stagnation lines, overcoming any remaining trap- Faradaic charging on one electrode (positive
ping force. In this way, it is possible to write bias) and capacitive charging on the other
and erase suspended particles, bacteria, or (negative bias), but a quantitative theory remains
microalgae on microelectrodes, as shown in to be developed and confirmed systematically by
Fig. 7a. This effect can be enhanced by adding experiments.
A 16 AC Electro-Osmotic Flow
AC Electro-Osmotic Flow, Fig. 7 (a) Collection of particle trapping by asymmetric polarization ACEO on
E. coli bacteria in tap water along the stagnation lines of electrodes with positive DC bias at higher voltage
ACEO flow on Au microelectrodes at low frequency (>3 V) (Reproduced from [7])
(100 Hz) and moderate voltage (1 V). (b) Preferential
References
fluid oscillates adjacent to a certain obstacle or an interaction between the air and the duct wall,
interface. So, as pointed out by Riley [1], the term a steady recirculating flow takes place within
acoustic streaming is misleading. Instead of the duct. The net effect is that dust or particles
acoustic streaming, he insisted on using steady accumulate at nodes.
streaming to encompass the case of incompress- On the other hand, researchers interested in
ible fluid. Acoustic streaming in this entry means mass transport in water waves have shown that
both of these effects. a similar phenomenon occurs for incompressible
Another important issue that must be flow (see [4] for an example). The salient feature
addressed is whether or not the Stokes drift flow is when an obstacle oscillates with high fre-
should be included in the streaming flow cate- quency in the quiescent viscous fluid. In this
gory. The Stokes drift flow is purely kinematic case the steady streaming emerges from both
and so it is basically different from the Eulerian sides of the obstacle along the oscillating direc-
streaming flow, which is driven by the time aver- tion at small viscosity [5]. Increased importance
age of the Reynolds stress term. However, the of the acoustic streaming in small scales has been
mass-transport effect given by the Stokes drift addressed in [6].
flow is not weaker than the Eulerian streaming. In this article, we will describe in some detail
On the contrary, in the case of a progressive the fundamentals of the three types of streaming
wave, the Stokes drift flow plays a dominant flows associated with the microfluidic applica-
role (e.g., application of flexural plate waves to tions. First, the quartz wind which corresponds
pumping and mixing in microfluidics [2]). There- to the one-dimensional compressible flow will be
fore, the Stokes drift flow must be understood as introduced. Second, the Eulerian streaming flow
a kind of streaming flow. in two-dimensional space will be considered.
Unfortunately, little is known about the histor- Kundts dust phenomenon will also be explained.
ical background of the study of acoustic stream- Finally, we will address the flexural plate wave
ing. Therefore, instead of trying to give the and its net effect, i.e., the Stokes drift flow.
historical background of the general concept, we
will briefly survey the modeling and applications
of acoustic streaming in microfluidics. A survey Basic Methodology
of steady streaming flow has been given by
Riley [1]. One-Dimensional Compressible Flow Model:
In the early period, study mainly focused on Quartz Wind
the acoustic streaming of compressible fluids. In this and next subsections, we closely follow
The simplest and seminal phenomenon associ- the review by Riley [1] to describe the physical
ated with acoustic streaming is the quartz wind. mechanism of acoustic streaming. We first con-
When the air experiences a high-frequency sider the case when the sound is generated from
oscillation given by an ultrasound source, a source and travels along a certain direction,
a progressive wave is established in the air. Due say the x*-direction, in a space. The governing
to the attenuation of the wave, a nonzero time- equation for the motion of the compressible
averaged Reynolds stress is built in the region fluid is
close to the sound source, and this stress pushes
the air in the direction of wave propagation. The @r @r u
0 (1a)
resultant wind is called a quartz wind. Lighthill @t @x
[3] has provided a physical and simple analytical
treatment of this steady streaming flow. Another @u @u
1 @p 4 @ 2 u
u n (1b)
important phenomenon is the so-called Kundts @t @x r @x 3 @x2
dust pattern. When the air within a duct receives
a standing wave, a nonzero time average of the where u* is the flow velocity, p* the pressure, t*
Reynolds stress is built inside the duct. Due to the the time, r* the density of the fluid, and n the
Acoustic Streaming 19 A
kinematic viscosity of the fluid. The fluid particle We then have
is assumed to oscillate back and forth with ampli-
tude A0, frequency o, and wavelength l. We then @ 2 u0 @ 2 u0 4 @ 3 u0 A
take 1/o, l/2p, A0o, and r0A0o2(l/2p) as the 2 d 0 (7)
@t2 @x 3 @t@x2
reference quantities for the time, length, velocity,
and pressure, respectively. Further, we expand The solution of this equation is
the density as
u0 exp2dx=3 cos x t (8)
r r0 1 e r0 e r1
2
(2) Notice that the basic mode is progressive. We
need the next higher-order equation of Eq. 1b,
where r0 corresponds to the undisturbed fluid which is
density. We also expand the dimensionless veloc-
ity and pressure, respectively, as @u1 @p1 4 @ 2 u1 @u0
d u0 (9)
@t @x 3 @x2 @x
u u0 eu1 e2 u2 (3a)
The right-hand side term can be understood as
a dimensionless force per unit mass. Substituting
p p0 ep1 e2 p2 (3b) Eq. 8 into the right-hand side gives
and
@u0
u0 exp2dx0 =3 cos x0 t ex1
@x xx0
(14)
1
eud eexp4dx0 =3 (15)
2
@u 1 2 @U @ue 1 @ 2 U @U @U
eu u p u (17b) e U V (21)
@t Re @t @t 2 @Y 2 @s @n
A
where u u, v and is the two-dimensional where ue(s) denotes the tangential component of
gradient operator. We use 1/o, L, A0o, and the potential flow velocity evaluated at the solid
rA0o2L as the reference quantities for the time, surface. We expand the velocity components as
length, velocity, and pressure, respectively. Note before
that the reference length L used here represents
a typical dimension of the obstacle. The Reynolds p
U, V U0 , 2=Re V 0
number Re defined as
p
oL2 e U 1 , 2=Re V 1 . . . (22)
Re
n
The leading-order equation is
is assumed to be large. For the small value of e,
we expand @U0 @u0e 1 @ 2 U 0
u, p u0 , p0 eu1 , p1 . . . (18) @t @t 2 @Y 2
The leading-order equation of Eq. 17b is then whose solution takes the following form
@U 1 @u1e 1 @ 2 U1 @U 0 @U0
U0 V0
where the complex functions f p must
satisfy the @t @t 2 @Y 2 @s @Y
continuity equation 17a and i 1 denotes the (24)
imaginary unit. Note that the inviscid region
governed by these equations comprises most of Here our interest is the time-independent flow.
the flow domain except the thin layer near the Taking the time average of the above equation
solid boundary. over one period of oscillation gives
Since the potential flow solution (Eq. 20) does
not satisfy the no-slip condition on the solid sur- 1 @ 2 U1 @U 0 @U0
U0 V0 (25)
face, we must consider a thin layer (called the 2 @Y 2 @s @Y
Stokes layer) adjacent to the surface n = 0, where
n refers to the local coordinate normal to the wall. The solution of this equation yields the streaming
In this thin layer, we use the strained coordinate velocity at the edge of the boundary layer,
Y defined as
" #
p 3 # du0e #
du0e
n 2=Re Y U 11 1 iu0e 1 iu0e
8 ds ds
We also use the velocity components U and (26)
V along the local coordinates s and n, respectively
(the coordinate s is along the surface). Then the where the superscript # denotes the complex con-
boundary layer equation becomes jugate. This velocity then acts as a boundary
A 22 Acoustic Streaming
1
u ud u p 2 u (27)
Res
Acoustic Streaming, Fig. 4 Eulerian streaming flow occurring between two parallel plates when a standing wave
takes place inside the duct. Distance between the nodes corresponds to a half wavelength of the standing wave
a Heater
IDT
Silicon
l
Thermopiles
Aluminium
Zinc oxide
Polysilicon
dM
Silicon
nitride
f
b l
particle trajectory
flexural plate A0
Acoustic Streaming,
wave
Fig. 5 Microfluidic device
propagation
utilizing the flexural plate B0
wave. (a) Fabricated h* y*
fluid
device; (b) shape of the
flexural plate and the x*
coordinates for analysis [2]
between the two. Here we derive the equations as is c = ol/2p. A particle attached on the flexural
well as the solutions for the Stokes drift flow plate describes an ellipse with major A0 and
given by the flexural plate wave. The case of minor B0 = rA0 rotating clockwise when the
one-dimensional compressible flow has already wave travels toward the right-hand side as
been treated in this article. shown in Fig. 5 [2]. The ratio r is given from
Consider a viscous fluid confined between r = pd/l, where d is the membrane thickness.
a flat wall at y = 0 and a flexural plate at A typical value of r is 0.1. The spatial coordinate
y = h. The flexural plate oscillates showing x is along the wave propagation direction and y
a traveling wave with a high frequency o normal to it.
by a well-established piezoelectric vibration As before we use 1/o as the time scale, l/2p as
system. Figure 5 illustrates the geometry of the length scale, A0o as the velocity scale, and
the flow model investigated in this entry. The rA0o2l/2p as the pressure scale, and then the
wavelength is l and the wave propagation speed dimensionless continuity and momentum
A 24 Acoustic Streaming
0.0025
Re = 4760
For instance, at Re = 4,760 we obtain hc = 0.041
which corresponds to hc = 0.65 mm at
0.002 l = 100 mm. This is very small compared with
the wavelength, but it is still far larger than the
0.0015 amplitude of the oscillation of the plate,
q
Re=1400
A0 = 6 nm.
0.001
0.0005
Re = 400
Key Research Findings
2f 0 2 Dm TSM Fabrication
Df 1=2 (1) TSM resonator fabrication usually utilizes photo-
Apiezo uq pq lithography [2]. The resonance frequency of the
Acoustic-Based Biosensors 31 A
quartz crystal depends on the shear modulus,
density, and material thickness [1]. After
polishing the quartz, photoresist is deposited, A
and the device is exposed to UV light through
a mask, which determines the active areas that
have to be metalized on the devices. On a TSM
resonator, they represent the location of the elec-
trode plates. A chemical interface is then added to
the remaining crystal surface, where molecules in
the liquid can bond to and be detected [2].
and are sensitive to their environment [8]. This noses mostly have the ability to detect very low
type of acoustic wave sensors consists of basic vapor concentrations [8]. If there is vapor on
components such as piezoelectric substrate the piezoelectric surface of the SAW device,
(crystals such as quartz, GaAs, or LiNbO3), the chemical substances in the vapor and in the
micrometallization patterns (electrodes), acoustic wave interact and the properties of the
interdigital transducers (IDT), and active thin wave changes. Sensitivity of the SAW detector is
films. Unlike BAW, which only interacts with dependent on the temperature of the piezoelectric
the environment at the opposite surface of the crystal and the vapor-pressure of the analyte.
material by traversing through it, SAW travels Chemical concentration of analyte of interest
along or near the surface of the piezoelectric can be tested by monitoring the frequency. The
material [3]. The piezoelectric device detects SAW chemical sensor requires no special support
small mass changes at the surface of the sensor gases and no consumables other than electrical
as a frequency response, as the electrodes on the power. Many of the SAW devices have been
surface of the piezoelectric substrate transmit and produced for military and defense applications.
receive acoustic waves [8]. The acoustic wave is They are used for on-the-spot vapor detection of
confined to the surface of the piezoelectric sub- plastic explosives containing nitro groups such as
strate, and the excited wave is propagated along trinitrotoluene (TNT) and cyclotrimethylenetri-
the crystal surface. The surface wave velocity nitramine (RDX) [8]. As a result, SAW sensors
changes due to the mass, or the viscosity changes are attractive for real-time detection and moni-
due to the binding reactions on the sensor surface. toring of toxic or hazardous gases and vapors, and
The range that SAW device operates depends on they are used in biology as a sensitive method for
the acoustic velocity of the crystal substrate and detection of biochemicals and biomolecular
IDT wavelength, and it usually lies between interactions [9].
lower MHz range to the GHz range [8]. These SAW devices operate at high frequencies,
devices are commonly used because of their abil- which lead to smaller, more sensitive devices
ity to directly sense the changes in mechanical that have limits in the picogram range [8]. SAW
and mass properties, and they have a broad range devices can be miniaturized by using photolitho-
of applications such as pressure, temperature, graphic techniques, and complex circuits can be
viscosity, mass, and chemical and biological present on the substrate surface. Miniaturization
sensing [8]. SAW sensors coated with organic makes them advantageous since the smaller
and/or inorganic materials can be used to analyze dimensions increases the sensing surface area
absorption on the surface of a device [9]. For and binding properties [6]. Moreover, there is
chemical sensing, the chemoselective material a dual-delay system with SAW sensors (Fig. 4).
must be tailored to the analyte of interest. During It is a layered structure and has three fundamental
its operation, SAW sensor responds to a gas or advantages. First, it compensates the influence of
vapor exposure in air by a shift in resonant signal temperature on the differential frequency and the
frequency and then recovers to the original signal air pressure. Second, it reduces the measured fre-
frequency, when the sensor is exposed to ambient quency of the system from MHz range to kHz
air free of the responding analyte [8]. The SAW range [6]. Third, SAW devices have the ability to
velocity changes according to the absorbed directly sense mass and mechanical changes in an
chemical substance, and it results in the alterna- environment, which makes them more sensitive
tion of the mass load, the elastic stress, and the than other electrical and thermal biosensors. Fur-
conductivity of the polymer coating on the SAW thermore, they have numerous advantages in bio-
device. logical applications [6]. These devices are also
SAW devices are also considered as elec- more cost-efficient than the optical biosensors
tronic noses. An electronic nose is a device and easier to handle. They are employed to study
that is composed of a chemical-sensing system biomolecular interactions and allow easy and
and a pattern-recognition system. Electronic direct detection of analyte of interest [8].
Acoustic-Based Biosensors 33 A
Acoustic-Based
f tr
Biosensors, Fig. 4 SAW
device dual-delay line fA
structure and its principal of MPc V Lw L
A
operation [24] VO
A A
f tr
f LiNoCB f0
f f0
f=f-f0
Bottom
Substrate
electrode
A
1. Thermal oxidation and etch 5. Ground/etch back and remove the box.
Acoustic-Based Biosensors, Fig. 6 Major steps of CMUT fabrication with wafer-bonding method (Reprinted with
permission from Ergun et al. [14])
not radiate significant amount of energy into liq- SH-APM sensors have also been applied in
uids, it minimizes damping in the liquid opera- immunochemical reactions. In order to observe
tion. Besides few exceptions, the SAW sensor antigen-antibody reactions through mass loading
induces considerable surface-normal displace- on the crystal surface, Dahint et al. [7] constructed
ment that radiates compression waves into the an immunosensor using a 150 MHz APM device
liquid and causes excessive damping. Since bio- on a ZX-LiNbO3 plate with a thickness of 0.5 mm.
sensors detect chemicals in liquid solutions, These sensors, which could detect mass as small as
BAW is often preferred over SAW for many to 200 pg/mm2 of crystal surface, marked a shift in
biosensing applications [2]. resonant frequency of around 1 kHz upon antigen
adsorption [7].
Biological Applications of SH-APM Sensors
SH-APM sensors have been utilized in various Biological Applications of TSM Resonators
biological applications. For example, Liren Many biosensors that employ TSM resonators are
et al. [15] presented a urease biosensor in which available on the market today. Depending on their
a 0.5 mm thick ZX-LiNbO3 substrate was used design, they can detect biological entities such as
and a resonant frequency approximately centered nucleic acids, viruses, bacteria, and cells. An
at 50 MHz. APM was generated and detected by example of TSM-based biosensor was presented
IDTs, with a period of 88 um and a center dis- by Davis et al. [16]. They used a coating of human
tance of 10 mm, on the same PZ crystal. The IgG and subsequently used a TSM resonator to
group also fabricated two pairs of IDTs on the detect the presence of sheep antihuman IgG as
substrate so that one would conduct biochemical marked by the decrease in the resonant frequency,
reactions while the other acted as a reference. signifying an increase in the mass loading from the
Using this device, they successfully measured attachment of the antibodies [16].
urea concentrations by detecting variations in TSM sensors can also be used as DNA-based
the resonant frequency [15]. sensors. Okahata et al. [17] studied the use of
A 36 Acoustic-Based Biosensors
Future Directions for Research wider areas due to two main limitations.
According to the radar equation, the
Advancements made over the past decades have radiofrequency signal changes with the fourth
culminated in the initiation and development of power of the distance between the radar unit and
acoustic wave biosensors. The ability to detect SAW sensor. As a result, high signal attenuation
biological molecules by these sensors may con- is governed by this equation. Second, only a few
tribute to treatment of many diseases such as HIV echo sounds can be distinguished because all
and cancer. In addition, these devices are prom- sensors in the detection range respond immedi-
ising in medical imaging given their ability to ately. Attaining this requires differentiated sen-
generate high-resolution pictures in real time. sors that respond within different time slots, and
However, there are still limitations to some of multiple sensors can be used with a limited num-
these devices that can be surmounted through ber of reflectors and limited identity-code
future research and though the potential of the capacity [25].
application of these devices to biological situa- In addition, generation of acoustic waves in
tions has been recognized, more work is left to be microstructures requires various processes and
done in order to employ them on a wider scale. technologies, with compatibility as one of the
Below are some limitations and potentials of the main limitations in the development of smart
devices. sensors. Restrictions of manual handling and fab-
rication process limit minimum thickness of the
SAW Devices devices to be about 100 microns. Adoption of
There are a number of potential limitations with silicon fabrication opens the possibility for the
the SAW sensors as they are today. One limita- integration of the acoustic devices and electronic
tion is that SAW sensors are sensitive to a variety driving circuitry, leading to the development of
of parameters [8]. In addition, the intensity of an smart sensors. This technique involves localiza-
acoustic wave can decrease in biological solution tion of specific fabrication steps to common areas
such that the sensing system may not work effi- to minimize the total number of process steps in
ciently. Furthermore, many SAW devices that are the fabrication process as well as to maintain
used in the gas-sensing applications may not be SAW and Lamb wave device integrity and
used to detect molecules in the liquid phase. quality [26].
When a fluid contacts the SAW surface, the lon- Most of todays acoustic wave generation and
gitudinal wave component pumps a compression focusing apparatus suffer from these further lim-
wave into the fluid [2]. In air, the energy losses itations: the piezoelectric arrays are expensive,
are not too effective. However, in many liquids, and it is hard to get high-level, well-distributed
all the energy is pumped out of the acoustic wave intensities. In addition, shocks are generated at
before it reaches the devices receiving trans- a rate of one or two shocks per second, whereas
ducer. SAW devices can perform well in liquids extracorporeal shockwave treatment (ESWT)
when they rely on surface waves with no longi- typically requires thousands of shocks per treat-
tudinal components like horizontally polarized ment. Acoustic lenses are mostly fragile and
shear waves, surface transverse waves, and the noneffective for larger apertures. Moreover, elec-
guided acoustic waves called love waves [2]. trohydraulic approach has disadvantages of
Recent advances in liquid sensors and immuno- nonuniform discharges, pain, and high noise
assays improved the detection limits. In addition, level.
surface technologies like functionalization with
fluorescent probes, DNA, and enzymes increased BAW Devices: TSM Resonator
the use of SAW devices in liquid samples The limitations of TSM resonators are mostly
[7]. There are also radar-type SAW sensors for related with the experimental setup, accuracy in
high-voltage applications that detect at long dis- modeling the sensor, and physical behavior of
tances. These sensors cannot be used scans of a viscoelastic polymer. Researchers should be
Acoustic-Based Biosensors 39 A
careful in model approximations, parameter esti- medium during nucleic acid analyses. Complex-
mations, and measurement errors. For acousti- ation of DNA may limit the flexibility of the
cally thin polymers, the thickness can be molecule strands and ionic strength changes or A
calculated with a small error, but the shear moduli interaction with the surface might vary extension
can hardly be extracted from measurements due and shape of the molecule. These may change
to negligible influence on the sensor behavior. On viscoelastic behavior and produce erroneous
the other hand, the coating thickness should be results from TSM resonators [4].
known because of the existence of multiple iden-
tical solutions for acoustically thick polymers CMUTs
[27]. In addition, for detailed interpretation of CMUTs have the potential to make ultrasound
sensor response in liquid requires more sophisti- a far more versatile and important imaging
cated experimental setup [4]. In addition, modality. This technology has made advances in
mechanical, optical, and electrochemical proper- acoustic matching and the microminiaturization
ties are helpful to characterize interactions at of electronics. It enables higher-frequency imag-
biological surfaces and interfaces. However, the ing, allows clinicians to view small features in the
needed equipment are expensive. While minia- body, and is useful for ultrasound imaging espe-
turized quartz arrays seem to present a solution cially in the area of volumetric in vivo imaging,
for low-cost sensor setups, miniaturization has its intravascular, and intracardiac research.
limitations because small electrode areas can
load enough molecules to achieve measurable
frequency shifts. As a solution, combination of Cross-References
automatized pipetting systems, microfluidics,
integrated sensor arrays, and multicomponent Biosensor
data analysis may open the way toward high- Biosensors Using Atomic Force Microscopes
throughput screening with acoustic wave Biosensors Using Lasers
sensors [4]. Biosensors Using Magnetics
Two major challenges are present in the appli- Biosensors Using Surface-Enhanced Raman
cation of TSM resonator as a biosensor. First, Scattering
cross-reactions such as unspecific protein adsorp- Piezoelectric
tion affect the accuracy of the results. This limits
the resonators use to binding sites that specifi-
cally bond to the targeted biological molecules
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Active Mixer 41 A
components within the micromixer itself, and thus turbulence does not occur. The Reynolds
e.g., magnetically actuated stirrers, or by the number of microfluidic flows in a conventional
application of an external force field, e.g., pres- straight microchannel is typically less than one, A
sure, ultrasound, acoustic, electrohydrodynamic, and the resulting Stokes flow is unidirectional.
electrokinetic, dielectrophoretic, magnetohydro- Even in curved or twisted microchannels, the
dynamic, thermal, and so forth [1, 2]. transverse component of the flow (i.e., the second-
ary flow) is very small compared to the streamwise
component since the inertial effects induced by the
Overview microchannel geometry are very weak. Therefore,
when the microchannels contain two or more fluid
Mixing occurs in many natural phenomena, streams, mixing of the different streams is domi-
including geophysical, ocean, and atmospheric nated by molecular diffusion effects. In microscale
flows, and is also an important step in industrial channels, the diffusive mixing process is very slow
processes involving chemical reactions, fermenta- compared with the rate at which the fluid is
tion reactions, combustion, and so forth. Tradition- convected along the channel. Typically,
ally, industrial mixing applications have always microfluidic flows have a high Peclet number,
been performed using large-scale apparatus. How- i.e., Pe = Ul/D, where U is the average flow
ever, in recent years, microscale devices velocity, l is the channel dimension, and D is the
(commonly referred to as microfluidic devices) molecular diffusivity. The diffusive mixing time is
have been proposed as a means of constructing given by tD ~ l2/D, and the mixing channel length
micro-total analysis systems (m-TAS) and lab-on- required to achieve an adequate species mixing
a-chip (LOC) devices. However, achieving a rapid increases linearly with the Peclet number
and efficient mixing of different reactants when (Lm
Pe l). For example, for an average flow
performing chemical and biological analyses in velocity of 500 mm/s, a channel dimension of
such microfluidic devices is highly challenging. 100 mm, and a molecular diffusivity of 10 mm2/s,
Therefore, the problem of developing enhanced the mixing time and length required to achieve
micromixing schemes suitable for microfluidic a complete species mixing are of the order of
applications has attracted significant interest 1,000 s and 0.5 m, respectively. However, many
within industrial and academic circles in recent practical biochemical applications, e.g., immuno-
years. As a result, the literature now contains assays and DNA hybridization, require the rapid
a large number of studies concerning micromixers mixing of macromolecules characterized by rela-
implemented using a variety of mixing strategies tively low molecular diffusivity. Therefore, the
[1, 2]. The micromixing schemes are categorized mixing performance obtained in typical
as either active or passive. The active mixing has microchannels is unsuitable for biochemical anal-
been defined in the above section. The passive ysis applications. Accordingly, a requirement
mixing refers to the mixing effect in systems exists for enhanced microfluidic mixing schemes
enhanced by virtue of their particular geometry to support the development of m-TAS and LOC
topologies, surface properties, or instability phe- devices. Typically, such schemes involve reducing
nomenon which occurs naturally under a static the diffusion length (i.e., l) and increasing the
(DC) electric field. The active mixing is easily interfacial contact area between the mixing
controlled using external mechanisms. Active species.
mixers are eminently suitable components for Although it is difficult to induce turbulence
reconfigurable microfluidic systems. (so-called Eulerian chaos) in microchannels, the
The characteristic size of the microchannels in mixing performance obtained in low Reynolds
microfluidic devices is very small (typically of the number flow regimes can be enhanced via the
order of several 100 mm), and hence the flow is chaotic advection mechanism (or so-called
dominated by viscous forces. As a result, the flow Lagrangian chaos). Chaotic advection occurs in
is restricted to the low Reynolds number regime, regular, smooth (from a Eulerian viewpoint)
A 42 Active Mixer
flows [3] and provides an effective increase in and by a momentum equation modified to include
both the interfacial contact area and the concen- the body force term, i.e.,
tration gradient by inducing a reduction in the
striation thickness and hence in the diffusion u 0 (1)
length. As a result, both the mixing time and the
mixing length are considerably reduced. In the and
case of a linear reduction in the striation thickness
@u
Basic Methodology rf p m2 u F: (3)
@t
dr
ur, t, (6)
dt Key Research Findings
where r is the location of the tracer particle and Many active mixing approaches have been pro-
u(r, t) is the Eulerian velocity field. The tracer posed in recent years. Broadly speaking these
particle position in the unsteady flow field at approaches can be classified in terms of whether
A 44 Active Mixer
Active Mixer,
Fig. 1 Pressure
perturbations induce lobe-
like distortions of the
interface and facilitate
rapid mixing in the device
with one pair of side
channel
or not they use moving parts to generate a mixing exponent data were applied as mixing indices to
effect. In those micromixers which do use mov- evaluate the intensity of the chaotic mixing
ing parts, mixing is commonly achieved using effect. The optimal operating parameters were
piezoelectric membranes, gas bubbles, and so then determined by plotting the Lyapunov expo-
forth. However, since the integration of these nent data against the amplitude and frequency of
moving parts within the overall microfluidic sys- the perturbation.
tem is challenging, and therefore expensive,
researchers have paid increasing attention to Electrohydrodynamic (EHD) Mixer
active micromixers containing no moving ele- In electrohydrodynamic (EDH) mixers, a mixing
ments. Numerous proposals for active mixing effect is created by applying an external electrical
approaches have been presented in recent years, field to a bulk flow containing both an electrical
and space constraints prevent their detailed conductivity gradient and a permittivity gradient.
discussion here. Hence, the following provides In the micromixer proposed by Moctar et al. [6],
no more than a high-level review of some of two fluids with identical viscosity and density but
the more fundamental techniques. For a compre- different electrical properties were injected into
hensive discussion of active mixing schemes, the mixing channel by syringe pumps, and the
interested readers are directed to the review electrodes were arranged in such a way that the
presented in [1, 2]. electrical field was perpendicular to the species
interface, as shown in Fig. 2a, creating a transver-
Pressure Perturbation Mixer sal secondary flow. The mixing effects induced
In pressure perturbation mixers, the species are by both DC and AC electrical fields were
perturbed by pulsing their velocities in some way. explored in a series of experimental investiga-
For example, Niu and Lee [5] presented an active tions. From Fig. 2b, c, d, it can be seen that the
mixer comprising a main channel and multiple mixing becomes more chaotic as the intensity of
side channels, in which the mixing species were DC electric field increases. The results revealed
stirred by pulsing the velocities of the fluids that the application of an appropriate voltage and
introduced through the side channels, as shown frequency to the electrodes yielded a satisfactory
in Fig. 1. The resulting stretching and folding of mixing performance within less than 0.1 s over
the fluid elements in the main and side channels a short mixing distance, even at Reynolds num-
induced a chaotic advection mixing effect. bers as low as 0.02.
A detailed particle simulation study was
performed to examine the respective effects of Electrokinetic-Based Mixer
the amplitude and frequency of the flow pertur- Oddy et al. [7] presented a micromixer in which
bations on the species mixing performance. AC electrical fields were used to induce the elec-
Poincare maps, constructed using the simulation trokinetic instability (EKI) phenomenon in
data, showed that for a given parameter set, the microscale channels at very low Reynolds num-
fluid flow exhibited both chaotic and quasi- bers, as shown in Fig. 3. EKI is similar to
periodic responses. Additionally, Lyapunov electrohydrodynamic instability, which can be
Active Mixer 45 A
Active Mixer,
Fig. 2 From [6] (a)
Configuration of
electrohydrodynamic A
mixer. Experimental
images of mixing (b) initial
condition, (c) after the
application of a DC electric
field of intensity
E = 4 105 V/m and
(d) E = 6105 V/m
Active Mixer,
Fig. 4 Numerical and
experimental images of
mixing for different
switching frequencies for
a constant driving electric
field of 100 V/cm
flow fields at various periods was analyzed by wire electrodes and the circular side-wall elec-
performing particle tracking simulations (i.e., trode and between the second wire electrode and
Poincare maps). The results indicated that the side-wall electrode, as shown in Fig. 5. The
a more chaotic mixing effect was achieved at particle tracing results revealed that the intensity
lower values of the switching frequency. of the chaotic mixing effect increased with
increasing T, resulting in a satisfactory mixing
Magnetohydrodynamic (MHD) Mixer result within 40 periods.
Various researchers have employed the magneto-
hydrodynamic (MHD) effect to implement
micromixers. For example, Bau et al. [9] devel- Future Directions for Research
oped an active micromixer which used either DC
or AC electrical and magnetic fields to generate It seems likely that future micromixer designs
Lorentz forces with which to stretch and fold the will continue the current trend for implementing
species fluids in a mixing chamber. The results species mixing without the use of moving
indicated that an adequate mixing result was mechanical parts. Furthermore, it seems reason-
achieved within several seconds. A more sophis- able to speculate that the use of electrokinetic
ticated MHD mixer was developed by Yi forces to induce a mixing effect will become
et al. [10] based upon the two blinking vortex increasingly common since this technique greatly
arrangement used to induce chaotic advection. simplifies the microfabrication process and
The proposed micromixer comprised a small reduces the cost and complexity involved in
cylindrical mixing chamber with an electrode embedding active mixers within microfluidic sys-
deposited on its side wall and two copper-wire tems. Electrokinetic forces have been widely
electrodes positioned eccentrically on the lower employed in active mixing schemes in recent
surface of the chamber. The chamber was orien- years, a case in point being the EKI mixer
tated such that its axis was parallel to the direc- presented by Oddy et al. in [6]. However, current
tion of an applied magnetic field, and a mixing EKI-based mixers suffer the drawback of a high
effect was generated by applying a potential dif- electrical voltage requirement. Accordingly,
ference of period T alternately between one of the low-voltage, AC electrokinetic techniques are
AFM 47 A
References
nFAD 1
I C (2)
d
Amperometric Detection, Fig. 4 (A) Illustration of the (b) is 40 mm in diameter. (C) Photograph of Bio-LSI chip.
cross section of a unit cell. The surface of the LSI was After wire bonding, the conductive wires were insulated
modified by metal (Au/Pt/Ti) and insulated by epoxy by PDMS. (D) Photograph of Bio-LSI chip unit. The
polymer (SU-8 3005) to define the electrode area exposed acrylic wall for the sample solution is set on the substrate.
to the solution. (a) Contact hole to expose Al pad. (b) (E) The photograph showing the experimental setting of
Sensing point. (B) Microgram of unit cells. Al pads (a) the Bio-LSI chip unit, RE and CE. (F) Complete view of
can be seen under the Au layer. The size of a sensing point the settings for the Bio-LSI system reproduce from [4]
analysis. Recently, LSI-based amperometric sen- Bio-LSI is a promising platform for a wide
sor has been demonstrated for bio-imaging appli- range of analytical fields, including diagnostics,
cation. Kumi Y. Inoue et al. developed an environmental measurements, and basic bio-
LSI-based amperometric sensor with 400 mea- chemistry. This chip has a mature manufacture
surement points as a platform for electrochemical technology and is able to be mass produced in
bio-imaging and multi-point bio-sensing [4]. The industry. Consequently, the LSI-based ampero-
system consists of a 10.4 610.4 mm CMOS metric sensor is promising and has great commer-
sensor chip with 20 20 unit cells. The whole cial interests for bio-detection and biochemistry.
size of the chip is shown in Fig. 4c. A very wide
current detection range is successfully achieved FET-Based Amperometric Sensor
from 1 pA to 100 nA from the systematic The field effect transistor is a nonlinear physical
level. Figure 4 shows the detailed structure of device which is able to work as an amplifier. By
the chip and experiment setup for concentration tuning the gate voltage, the current change
imaging. between drain and source can provide an ampli-
The OsHRP-coated LSI was spotted with fied signal to characterize the changes of gate
glucose oxidase and used for bioelectrochemical voltage. By inserting an amperometric sensor
imaging of the glucose oxidase (GOx)-catalyzed between the drain and source, the information at
oxidation of glucose. Figure 5 shows the transi- the gate is recorded and analyzed. The commer-
tion of the redox current-based images of H2O2 in cial metal-oxide-semiconductor field effect tran-
the flow streams with the OsHRP LSI chip. sistor (MOSFET) has been used to design
When the H2O2 reached the chip, H2O2 was oxi- microfluidic sensor for particles counting. Xu
dized by HRP followed by the oxidation of Os3+ D. et al. has developed the MOSFET-based par-
to Os2+. The reduction current of Os3+ to Os2+ at ticle counter which can provide a current modu-
the 400 measurement points can be monitored by lation with ultrahigh sensitivity, due to its
a real-time image with a color scale at 400 points. nonlinear amplifying property [5]. The detection
Figure 5 shows clear and rapid changes of H2O2 limit of volume changes can reach to 0.006 %. In
flowing over the LSI chip. recent years, the electrical properties of graphene
A 52 Amperometric Detection
Amperometric Detection, Fig. 5 Color maps indicat- point, a reduction in current was observed according to
ing the current intensities of 400 electrodes obtained by the Os3+ generation by an HRP reaction toward H2O2. The
the 3 s injection of 10 mM H2O2 on the OsHRP modified time-dependent transition of the maps is shown in
Bio-LSI chip in PBS flow stream. An inlet and outlet of the 16 images. The operational potential was set to 0.0 V
flow were set in the upper left and lower right directions of versus Ag/AgCl/sat.KCl reproduce from [4]
the figure, respectively. When H2O2 reached a sensing
have been extensively studied, and graphene- microfluidic systems on glass or flexible PET
based sensor with good sensitivity has been substrates, which were used to detect flow veloc-
developed for many bio-detection applications. ity in the microfluidic channel as shown in Fig. 6
Due to the fact that graphene has a 2-D structure [6]. The counter-ions within the EDL will move
and every atom is exposed on its surface, the simultaneously under the pressure-driven fluids
electrical properties of graphene are very sensi- flow, which leads to a convection streaming cur-
tive to the change of the charge environment rent or streaming potentials between the two ter-
induced by the reaction or adsorption of analytes minals of the microfluidic channel. So the fluid
on the surface. Based upon the merits, graphene is flow velocity can be detected by characterizing
a promising material for highly sensitive sensors. the streaming current or streaming potentials with
The graphene-FET-based amperometric sensor is a SGGT integrated in the microfluidic channel.
able to provide even higher sensitivity due to the Figure 6a shows the schematic structures of
nonlinear properties of FET which has been a solution-gated graphene transistors on a sub-
widely used in the sensing of gas molecules, strate, where the graphene film is partially
DNA, and flow velocity. covered by a poly(dimethylsiloxane) (PDMS)
For example, Rong Xiang et al. has developed channel. One Ag/AgCl wire was injected into
solution-gated graphene transistor (SGGT) in the microchannel as the gate electrode.
Amperometric Detection 53 A
Amperometric Detection, Fig. 6 (a) Schematic dia- characteristics (IDSvs VG, VDS = 0.05 V) of a SGGT
gram of a SGGT integrated in a microfluidic chip. (b) in a flexible microchip filled with static 1 mM KCI solu-
Raman spectrum of single-layer graphene on a glass sub- tion at different bending status. (df) The photographs of
strate. The insert shows the photograph of the graphene in the flexible microchip at different bending status (d) flat,
the microfluidic chip. Scale bar is 0.5 mm. (c) Transfer (e) outcurve, and (f) incurve reproduce from [6]
The large-area single-layer graphene can be syn- with the flow velocity stepping up gradually.
thesized on copper foil with the chemical vapor The channel current increased with the increase
deposition (CVD) method. The graphene film of flow velocity. If flow direction is changed from
was then transferred onto a glass substrate or the gate to the graphene, the current profiles will
flexible PET substrate (Fig. 6df). The Raman be as shown in Fig. 3c. It is observed that the
spectrum of the graphene film indicates that the channel current IDS decreased with the increase
graphene film is approximately single layered as of flow velocity. The current changes of the
shown in Fig. 6b. The channel length of device for either flow direction were shown in
a graphene transistor between source and drain Fig. 3d. Since the current changes resulted from
electrodes is 1 mm, which is slightly wider than the modulation of the effect gate voltage by the
the width of microfluidic channel (0.8 mm), as convection steaming potential induced by
shown in Fig. 1b. pressure-driven flows, the shift of the effective
The experimental result of the device is shown gate voltage under different flow velocity was
in Fig. 3 with Ag/AgCl gate electrode set at extracted according to the current change shown
fixed gate and drain voltages (VG = 0.15 V, in Fig. 3d, which shows an approximated linear
VDS = 0.05 V) in alternate static and flowing relationships between the shift of the effective
of 10 mM KCl solution. Figure 3b shows the gate voltage, and the flow velocity for both flow
channel current profiles as a function of time directions. Consequently, by calibrating the
A 54 Amperometric Detection
Amperometric Detection, Fig. 7 Time-dependent is from graphene to Ag/AgCl gate electrode. (c) The flow
channel current of a SGGT characterized in alternate static direction is from Ag/AgCl gate electrode tographene. (d)
and flowing KCl solution with different velocities. The change of channel current and the shift of effective gate
VG = 0.15 V, VDS = 0.05 V. (a, b) The flow direction voltage as functions of flow velocity reproduce from [6]
device with standard fluids flow, the device can electrochemical properties of the material such
provide very high sensitivity for characterizing as graphene, SWNT, etc., are expected to further
the flow velocity (Fig. 7). develop to overcome the detection limits. For the
The ultrahigh detection sensitivity enables the second aspect, parallel analysis of multiple bio-
graphene promising materials for amperometric logical samples can provide rapid detection plat-
sensor in the lab-on-a-chip applications. The form which is a promising technology in the point
graphene-based sensor is now at its early stage of of care diagnosis.
invention, and lots of new graphene-based amper-
ometric sensors with stable performance are
expected in the near future. On the other hand, the Cross-References
mass production of graphene with good quality
also remains to be enhanced in future development. Amperometric Techniques
Future work shall be focused on two aspects: (1) 1. Christian N et al (2012) Implantable enzyme ampero-
metric biosensors. Biosens Bioelectron 35:1426
increase of the sensitivity of the amperometric
2. Bard AJ, Faulkner LR (1980) Electrochemical
microfluidic sensor and (2) multiple analysis of methods: fundamentals and applications. Wiley,
biomolecules. For the first aspect, new New York
Amperometric Techniques 55 A
3. Ney H et al (2009) Fabrication of a multichannel (or working) electrode is placed at the end of
PDMS/glass analytical microsystem with integrated the separation stream so as to permit detection of
electrodes for amperometric detection. Lab Chip
9:115121 sample components as they emerge and pass over A
4. Kumi Y et al (2012) LSI-based amperometric sensor the electrode surface. The applied potential must
for bio-imaging and multi-point biosensing. Lab Chip be sufficient to cause facile oxidation or reduc-
12:34813490 tion of the target analytes; and, in general, the
5. Xu D et al (2007) Ultrasensitive fluidic sensors with
amplifications from both fluidic circuits and magnitude of the measured current is directly
MOSFETs. Appl Phys Lett 91:013901 proportional to the analyte concentration.
6. He R et al (2012) Solution-gated graphene field effect
transistors integrated in microfluidic systems and used
for flow velocity detection. Nano Lett 12:14041409
Overview
species, oxidized or reduced at the surface of the coated onto the inner walls) for HPLC or differ-
sensing electrode and therefore is directly pro- ential electrical migration of the charged analytes
portional to the concentration of the molecule toward the electrokinetic drive electrodes for
detected as defined by Faradays law (Eq. 1). CE. Many of these separation techniques are
being downsized using microfabrication
dQ dN methods, referred to collectively as microfluidics.
it F (1)
dt dt
!
0:059 aw ad
E E0 log C Db (3)
j nj aaA aB
Analyte Sample
Flow Direction
Detection
Current A
Amperometric Techniques, Fig. 2 Schematic of amperometric detection cell (general representation for lab-on-a-
chip (LOC) applications)
A 58 Amperometric Techniques
iRu Rc
Z FW C DLW i =0 Z FA C DLA
Amperometric Techniques, Fig. 3 Electrical model of the auxiliary electrode; CDLA is the double-layer capaci-
three-electrode cell (ZFW is the faradaic impedance at the tance at the auxiliary electrode; RC is the resistance of the
working electrode; CDLW is the double-layer capacitance electrochemical cell; Ru is the uncompensated resistance;
at the working electrode; ZFA is the faradaic impedance at and i is the current)
Amperometric a
Techniques,
Fig. 4 Schematic of EC
electrode placement for the Working
Electrode
different detection
electrode configurations:
(a) end-channel detection; b c
(b) in-channel detection;
and, (c) off-channel
Working
detection Electrode Decoupler Working
Electrode
well-defined analyte plugs maintained within the migrating analyte so as to maximize the delivery
capillary immediately begin to spread radially of the sample to the detection electrode surface.
upon exiting and moving into the much larger
volume detection reservoir. Therefore, care
must be taken that the detection electrode, in Key Research Findings
this case, is located close to the exit of the chan-
nel, typically 25100 mm, but not so close as to The first microfluidic lab-on-a-chip (LOC)
interact unduly with the residual CE voltage that devices were reported by Harrison and Manz in
does extend slightly beyond the separation chan- the early 1990s. These devices consisted of sim-
nel. A detailed study of the residual post-capillary ple channels, several cm in length and 10100 mm
CE field, and the effect of end-channel electrode in width and depth, etched into glass microscope
placement, has been reported by Klett slides or silica plates via conventional
et al. [9]. The in-channel approach has been micromachining techniques [10]. In this work,
employed by researchers; however, this approach samples were injected onto the chip and moved
is limited due to bubble generation in the channel, as required through the channel network by elec-
which can significantly decrease the signal-to- troosmotic flow; separations were carried out by
noise (S/N) ratio of the electrochemical signal, electrophoretic mechanisms common to bench-
as well as the high electric field can significantly scale CE instrumentation; and target analytes
effect the resting background current generated at were detected and quantitated via fluorescence
the surface of the working electrode. detection. Subsequent studies by these and other
In regard to electrode geometry, the EC groups demonstrated unique advantages for the
detector response is proportional to electron lab-on-a-chip approach: high separation efficien-
generation (or consumption) during the desired cies, short analysis times, minute sample and
redox reaction. Therefore, increasing the area of reagent consumption, high sample throughput,
the detection electrode will increase the detec- and easy automation.
tion signal. The tradeoff is that the background In 1996, the first use of amperometric detection
noise increases linearly as well. Due to the con- in microchip CE devices was reported by Wooley,
fined geometry of the microchannel, in-channel Lao, Glazer, and Mathies [11] who used off-chip
electrodes are limited to simple rectangular CE-driving electrodes together with a photolitho-
shapes determined by the channel width and graphically patterned Pt electrode placed just out-
exposed electrode area. This configuration is side the CE channel for on-chip detection of
sufficient to maximize the contact of the analyte catechol neurotransmitters by direct oxidation
with the electrode surface. For an end-channel and DNA fragments and PCR products indirectly
electrode configuration, however, a simple rect- by EC monitoring of an electroactive intercalating
angular planar electrode is not the only option. agent. In another end-channel approach, Baldwin
The conventional planar geometry can be and colleagues [12] were the first to demonstrate
altered to match the shape of the radially a lab-on-a-chip device that incorporated both CE
Amperometric Techniques 61 A
high-voltage electrodes and all amperometric EC electrode) and allow the EC-working electrode
detection electrodes directly onto an integrated, to be placed in-channel where higher efficiency
microchip platform. Key features of their device separations can be realized. Similarly, Lai A
were the formation of a shelf or ledge at the end et al. [16] integrated a Pd-film decoupler with
of the separation channel to restrict the height a series-dual electrode to significantly enhance
of the detection reservoir near the working the oxidation current when switching between
electrode and the radial pattern of the working the single electrode mode and dual-series mode.
electrodes. These features enabled optimization In another work, Wu et al. [17] electrodeposited
of EC detection by decreasing the electrical resis- Pt nanoparticles on a Au ground electrode to
tance and CE voltage drop in the detection region construct the decoupler and achieved a limit of
and increasing the residence/contact time of the detection of 0.125 mM at a S/N = 4 for dopamine
analytes with the working electrode, respectively. with a Au working electrode.
Another novel CE microchip end-column Compared to alternative detection approaches,
approach has been reported by Ertl et al. [13], amperometric methods are ideally suited to
which integrates sheath flow into the CEEC sys- microfabrication processes since the electrodes
tem to increase analyte velocity in the detection themselves have the ideal dimensions
reservoir as it exits the CE separation channel. (110 mm) for incorporation onto a microchip.
Their study further elucidated the importance of Most importantly, patterning and construction
the spacing between the working and reference of EC microelectrodes are extremely compatible
electrodes, with optimum detector performance with mainstream microfabrication technologies
achieved for a spacing of
20 mm. Using an such as photolithography and sputter-coating.
electrically isolated potentiostat with a floating As a result, electrodes (and electrochemical
working electrode, Martin and coworkers [14] cells) of different shape, size, location, and
successfully detected catechol with minimum composition can be patterned directly onto
band-broadening effects by placing the working glass, silica, and other substrates with a high
electrode just inside the separation channel but degree of accuracy using the same photolitho-
without any decoupling. Off-channel EC detec- graphic techniques employed to fabricate the
tion approaches have been also explored in micro- microchips CE channels, thereby allowing the
chip platforms. For example, the Lunte groups detection operation to be fully integrated onto
demonstrated two different methods for the chip (Fig. 5). In addition, multiple electrodes
decoupling the CE field from the EC detection in different CE channels or multiple electrodes
system. In the first, decoupling was achieved by with different response characteristics can be
laser-cutting a series of 20, 30 mm holes in the roof created to allow the device to fit the intended
of the CE channel into the upper glass cover plate application optimally (Fig. 5b). This level of
and then filling them with cellulose acetate [8]. In integration does not exist with other detection
the second, a 500-mm-wide Pd decoupler strip was approaches and has several performance and
fabricated via photolithography and sputter- application advantages. For example, with LIF,
coated onto the glass substrate that formed the multichannel detection requires either the use of
bottom of the CE channels [15]. When placed multiple fluorescence systems or a means of scan-
directly in the CE channel and used as the CE ning a single fluorometer across multiple spots on
cathode, the Pd was able to absorb the H2 gas that the chip.
is generated during normal CE operation and Chip-based amperometric EC detection tech-
leads to the formation of bubbles in the channel. niques have been utilized in several noteworthy
The key advantages of these approaches analysis situations most commonly in clinical
(and others like them) are that they allow micro- (e.g., immunoassays and protein analysis) and
chips to operate at higher field strengths (up to environmental sensing. A recent review by
1,700 V/cm) with superior limits of detection Vandaveer et al. [19] provides an excellent over-
(
25 nM for dopamine with a carbon fiber view of these applications.
A 62 Amperometric Techniques
Amperometric Techniques, Fig. 5 Examples of differ- electrodes (Reprinted with permission from Woolley
ent electrode geometries that can be fabricated via photo- et al. [11]) and (b) multiple radial working electrodes
lithographic techniques: (a) rectangular working (Reprinted with permission from Keynton et al. [18])
Cross-References Cross-References
Anisotropic Etching
Synonyms
Definition
Different etchants used in anisotropic wet etch silicon due to their compatibility with
etching have specific etch rates for each crystal- metals involved in on-chip circuitry, whereas
lographic plane in the material being removed. It alkali hydroxide etchants (NaOH, CsOH) are
is the difference in etch rates between planes used for their faster etch rates [3]. A commonly
that produce shapes within the material being used alkali hydroxide etchant for silicon is
etched that appear to follow the planes within potassium hydroxide (KOH) which provides
the crystalline structure as seen in Fig. 1. For a high level of selectivity between h100i and
more information on the crystalline structure h111i planes. Materials that are highly resistant
of silicon and how miller indices are used in to these etchants, such as silicon nitride and
designating crystal planes, see The MEMS silicon dioxide, are often utilized as etch stops
Handbook [1]. It is commonly accepted that in more complex designs. Detailed specifica-
the h111i plane etches the slowest regardless of tions for each etchant including etch rates in
the etchant used. However, the crystal plane that specific crystal planes and recommended tem-
etches the fastest depends on the etchant com- peratures, concentrations, and etch stops are
position [2]. Ammonium hydroxide etchants available in the literature such as those of
(NH4OH and TMAH) are frequently used to Nguyen and Wereley [2].
a b
c d
Anisotropic Etching,
Fig. 2 Examples of
undercutting that is
possible for (a, b)
odd-shaped masks and
(c, d) misaligned masks
relative to crystal planes
Anisotropy 67 A
Careful design and alignment of the mask trench bottom, exposing the substrate to the
must be ensured when involving directional chemical etchant [4]. Several companies spe-
dependence in the etching process. Because the cialize in the production of microfabrication A
planes will continue indefinitely until they reach equipment for various reactive ion etching pro-
a surface or another plane, undercutting of the cesses. These processes, commonly referred to
masking material is possible. A shape that does as deep reactive-ion etching (DRIE), are capable
not align with the correct planes can produce an of producing channels with a depth on the order
opening that circumscribes the original shape if of 500 mm and aspect ratios on the order of
etched long enough as seen in Fig. 2. The angle 30:1 [5].
produced between the slow-etch plane and the
surface after etching is dependent on surface
orientation and the crystalline structure of the Cross-References
material. Such angles are found in the literature
and are utilized in design to achieve accurate Bulk Micromachining
depths and specific slopes (including vertical). Dry Chemical Etch
Equipment used in wet etching can be readily Fabrication of 3D Microfluidic Structures
found in most laboratories. The material being Isotropic Etching
etched is suspended within a beaker containing Plasma Etching
the etchant while a magnetic stirrer ensures con- Reactive Ion Etching (RIE)
tinuous transport of reactants to and products Silicon Micromachining
from the surface of the material. A simple Sputter Etching
temperature-controlled hotplate regulates the Wet Chemical Etch
optimum temperature.
Anisotropic dry etching is a method for cre-
ating high aspect ratio trenches with nearly ver- References
tical walls. The two common techniques utilize
a protective coating along the vertical walls of 1. Gad-el-Hak M (2002) The MEMS handbook. CRC,
Boca Raton
an etched trench such that the bottom of the
2. Nguyen N, Wereley S (2002) Fundamentals and appli-
trench etches at a faster rate allowing for large cations of microfluidics. Artech House, Norwood
aspect ratios. Each is independent of crystal 3. Lobontiu N, Ephrahim G (2005) Mechanics of
orientation. The first method forces the reactant microelectromechanical systems. Springer, New York
4. Franssila S (2004) Introduction to microfabrication.
gases to condense on the sidewalls of the trench
Wiley, Chichester
by lowering the temperature of the materials 5. Tabeling P (2006) Introduction to microfluidics.
with liquid nitrogen. Ions are then accelerated Oxford University Press, Oxford
toward the trench by means of an electric field.
This physical shower of ions results in an etch-
ing effect. After ion bombardment removes the
bottom layer, it can be etched further while Anisotropy
the sidewalls are protected by the cryogenically
frozen inhibitor. Another novel approach, Definition
commonly referred to as the Bosch process
(named after its inventor, Robert Bosch), alter- The directional dependency of a measurable
nates additive and subtractive microfabrication property of a material.
techniques. During the additive stage, chemical
vapor deposition is used to coat the sidewalls
of the trench with a polymeric passivation Cross-References
layer. Similar to the previous method, ion
bombardment is then used to sputter off the Sputtering for Film Deposition
A 68 Anode
Anode
Glass
Cathode
Definition +
UB
Cross-References
Anodic Bonding, Fig. 1 Schematic setup for anodic
Electrophoresis bonding of silicon to glass
T = 400 C A
Current
T= 350 C
Time
Anodic Bonding, Fig. 2 Current profiles during anodic Anodic Bonding, Fig. 3 Microreactor chip assembly by
bonding at different temperatures anodic bonding of silicon and glass
attractive force then pulls the surfaces into direct cavity can occur. This may have detrimental
contact, which explains why the bonding always effects in some device applications [3].
starts at the point contact of electrode and spreads The bonding strength for silicon-glass anodic
radially. The radial spreading of the bonding front bonding is measured typically at 1020 MPa.
makes it impossible for air to become trapped Some have reported between 30 and 40 MPa at
between the bonding surfaces. The electrostatic higher bonding voltage [2]. The pull test has
force depends on a number of factors, such as the demonstrated that bonding strength is higher
total applied voltage, the width of the gap, the than the fracture strength of the glass, i.e., the
concentration, and the distribution of ions in break in the pull test occurs only in the glass and
the glass, whether under the high field the negative not at the silicon-glass interface.
ions will drift out of the glass or the positive ions Anodic bonding of glass and silicon has
will drift into the glass and possible formation of found many applications in microsystems,
surface charge on the glass (the effect of such MEMS, and microfluidics, including the fabrica-
charge would partially shield the charge in the tion of pressure sensors, accelerometers,
glass, thus reducing the field in the gap). micropumps, and other fluid handling devices.
The chemistry of anodic bonding is an oxida- A typical microfluidic device is shown in Fig. 3,
tion process at the interface. Due to the high elec- which is a microreactor chip to perform chemical
trostatic field, oxygen anions leave the glass and synthetic reactions [4]. The microreactor was
create Si-O-Si bonds, i.e., a thin layer of SiO2 is made by wet or dry etching of silicon to form
formed. This gives the strong bond between sili- microchannels and sealed by anodic bonding of
con and glass, and the bonding becomes irrevers- a glass slide on top with inlet and outlet holes.
ible. The formation of thin SiO2 layer at the Anodic bonding can provide an airtight seal to
bonding interface has been verified by Rutherford ensure proper function of fluidic flow in the
backscattering spectroscopy. It has also been dem- microchannels. Silicon is a preferred material in
onstrated that the rate of oxygen release is directly microfluidic systems because micromachining of
proportional to the average current passing silicon is the most mature fabrication technique.
through the bonding area. In areas where silicon Complex structures can be made in silicon by
and glass are in direct contact due to electrostatic either wet chemical or reactive ion etching.
attractive force, the oxygen anions will oxidize the Anodic bonding of silicon with glass can also
silicon into SiO2 and form the anodic bond. The help in the building of multistack microsystems
oxygen is therefore totally consumed. However, in to perform complicated functions. Figure 4 shows
areas where silicon is not in direct contact with an example of a micropump made by silicon-glass
glass, for example, in the case of an etched cavity micromachining and anodic bonding [5].
into the glass or silicon, oxygen evolution into the This is a glass-silicon-glass multistack system.
A 70 Anodic Bonding
Anodic Bonding,
Fig. 4 Schematic cross Pyrex
section of a micropump
assembled by anodic
bonding of glass-silicon- Actuation cavity Poly-Si
Contacts
glass
Si
Pumping chamber
Output
Pyrex
Input Microvalve
The top Pyrex glass provides a hermetically sealed The transparency of glass allows optical
cavity so that the polysilicon heater can heat up the inspection or detection of fluidic flow/reaction
air in the cavity. Expansion of the hot air in the in microfluidic devices. It also facilitates
cavity will compress the silicon membrane to per- alignment of microstructures in glass with
form a pumping action in the pumping chamber. microstructures in silicon.
The bottom Pyrex glass seals the pump chamber as Although glass-silicon sealing or assembly is
well as provides the input and output holes for the most common form of anodic bonding, the
fluid flow in and out of the pump chamber. technique can also be applied to the sealing of
Anodic bonding is the primary method for many metals with a variety of glasses. Successful
packaging silicon microstructures, for example, seals have been made with borosilicate glass,
packaging of pressure sensors, accelerometers, soda lime glass, potash soda glass, aluminosili-
and solar cell panels, because of its relatively cate glass, fused silica, and fiber optical glass
low bonding temperature. With bonding temper- [1]. Strong seals have also been made to some
ature being below the glass transition point, the ceramic materials. For those metals and alloys
glass thermal expansion coefficient remains con- whose thermal expansion coefficients are
stant, which enables stress-free packaging of sil- matched to the above glasses, successful anodic
icon microstructures (provided the glass has bonding has also been made, such as tantalum,
a good thermal expansion match with silicon titanium, Kovar, Niromet 44, and to semiconduc-
over the process temperature range). In addition, tors such as germanium and gallium arsenide, in
with the low bonding temperature, there is no addition to silicon [1].
measurable flow of glass, which ensures sealing
around silicon microstructures without any loss
of dimensional tolerances. Apart from low bond- Basic Methodology
ing temperature, there are other attractive fea-
tures of anodic bonding: Although the principle of anodic bonding is sim-
Since glass is an electrical insulator, parasitic ple, successful bonding depends on appropriate
capacitances are extremely small. selection of bonding materials, good preparation
The bonding process can be readily performed of bonding surfaces, optimization of bonding
in vacuum, allowing hermetic sealing or zero conditions, and inspection of bonded samples.
pressure reference cavity to be formed These are explained in the example of silicon-
(or sealing with special gas mixtures). glass anodic bonding.
Anodic Bonding 71 A
Material Selection to this standard. Although anodic bonding is fairly
Silicon material is normally of standard wafer tolerant to particles and contaminations on the
sizes and thicknesses. As the anode part of the bonding surfaces, they still need to be cleaned A
bonding pair, the silicon should be low in resis- prior to anodic bonding, unless the wafers are
tivity. However, some applications require an straight out of the box. A study has demonstrated
insulating layer to prevent electrical leakage in that an H2SO4 + H2O2 acidic clean of bonding
silicon. In this case, a thin layer of silicon dioxide surface can significantly reduce the bonding time
or silicon nitride should be deposited on the sili- compared to a simple cleaning with acetone
con surface. The insulating layer does not prevent [6]. Thorough cleaning is even more important
anodic bonding of silicon with glass. However, for structured wafers. Whether the bonding sur-
a thicker insulating layer can increase the bond- faces are hydrophobic or hydrophilic can also
ing time dramatically. For example, for a 0.5 mm- have an influence on bonding. At low temperature,
thick oxide layer, a bonding time of approxi- a hydrophilic surface is easier to bond than
mately five times longer is required [2]. For a hydrophobic surface. However, such difference
anodic bonding with glass, p-type silicon wafers disappears at high bonding temperature [7].
(boron doped) are preferred, as a high electro-
static field is much easier to build up, particularly Process Parameters
at low temperature, at the silicon side of a silicon- The basic process parameters for anodic bonding
glass interface for p-type silicon than for n-type are temperature, voltage, clamping pressure,
silicon (phosphor doped) [6]. However, for high- environment, and bonding time.
temperature anodic bonding (>400 C), the dop- Bonding temperature is normally in the range
ing type of silicon is no longer a factor. 300500 C. Higher temperature promotes
Borosilicate glasses such as Corning 7740 better mobility of positive ions in glass,
Pyrex glass, Schott Borofloat 33 glass, and resulting in a faster bonding process. How-
Hoya SD-2 glass are commonly used for anodic ever, the temperature must be below the soft-
bonding with silicon. They are of almost the same ening point of glass (glass transition
coefficient of thermal expansion (CTE) as that of temperature).
silicon. They also have the necessary electrical Typical bonding voltage is in the range of
conductivity at the bonding temperature. How- 2002,000 V. High voltage helps to drive the
ever, some special applications require the glass positive ions in glass toward the cathode, leav-
to be high in resistivity (Corning 7070 glass) or ing a negatively charged region at the bonding
high in spectral transmission (Pilkington CMZ interface. The electrostatic force at the inter-
glass). Glasses that have a CTE different from face pulls the silicon and glass in direct con-
silicon or a low concentration of mobile positive tact, resulting in oxygen ions in glass
ions are still able to be anodically bonded with oxidizing the silicon to form strong chemical
silicon, but with special process conditions. For bonds. Figure 5 shows the bonding time as
example, to bond Corning 7070 glass with sili- a function of bonding voltage for anodic bond-
con, much higher temperature and voltage are ing of Corning 7740 glass with silicon [6]. It is
needed. To bond a Fisher glass slide whose CTE apparent that higher voltage results in a shorter
is very different from that of silicon, the bonding bonding time. For glasses with low concentra-
temperature has to be much lower (150 C), and tion of sodium ions (Na+), both high tempera-
bonding time is much longer (60 min) [6]. ture and high voltage are necessary for
a successful bonding. Higher voltage is also
Preparation of Bonding Surfaces needed for thicker glass. High voltage also
A well-polished surface is essential for a successful helps to reduce the temperature necessary for
bonding. The average surface roughness (Ra) a good bonding.
should be less than 5 nm. Commercial silicon Standard silicon wafers are not completely flat
wafers and glass substrates are normally polished but with 2040 mm bowing and wrapping.
A 72 Anodic Bonding
where ea is the permittivity of air, V is the applied Anodic bonding is a fairly mature technique.
voltage for anodic bonding, Eeff is a material Many basic issues have been dealt with. Future
constant which characterizes the elastic stiffness researches will mostly focus on widening its appli-
of the materials, a is the half width of the channel, cations, particularly in microelectronic and
and d is the channel depth. A similar problem microsystem (MEMS) packaging. Packaging can
happens when packaging pressure sensors, significantly influence the performance of final
micropumps, and accelerometers by anodic devices and systems. Therefore, anodic bonding
bonding. Most of these devices have has to be integrated as part of the manufacturing
micromachined silicon membranes, diaphragms, process, and its influence should be fully taken into
or suspended beams. These micromechanical account at the design and manufacturing stages.
structures may stick to the glass when they are
too flexible and too close to the bonding interface.
Although anodic bonding is a low-temperature
Cross-References
process compared to fusion bonding which
requires temperatures up to 1,200 C, the bonding Wafer Bonding
process can still induce thermal residual stress in
the silicon-glass assembly, causing bowing of the
bonded wafer. The thermal expansion of Pyrex
References
glass is not a constant but changes with tempera-
1. Wallis G, Pomerantz DI (1969) Field assisted glass-
ture. The higher the bonding temperature, the metal sealing. J Appl Phys 40(10):39463949
larger the mismatch between silicon and glass. 2. Obermeier E (1995) Anodic wafer bonding.
For Corning 7740 Pyrex glass, the crossover Electrochem Soc Proc 957:212220
3. Roger T, Kowal J (1995) Selection of glass, anodic
point with silicon (where the thermal expansion
bonding conditions and material compatibility for
of silicon is exactly the same as that of Corning silicon-glass capacitive sensors. Sens Actuator
7740 glass) is around 300 C. Experiments A4647:113120
showed that bonding made at this temperature 4. Cao E, Gavriilidis A, Cui Z (2002) Application of
microengineered reactors in chemical engineering.
can be maintained flat within 5 mm [3]. The
MST News 3(02):1416
crossover point is also dependent on the thickness 5. Acero MC, Plaza JA, Esteve J, Carmona M, Marco S,
of glass. Experiments indicated that for 0.5 mm- Samitier J (1997) Design of a modular micropump based
thick glass (Corning 7740), the crossover temper- on anodic bonding. J Micromech Microeng 7:179182
6. Lee TMH, Lee DHY, Liaw CYN, Lao AIK, Hsing IM
ature is at ~315 C, while it becomes ~260 C for
(2000) Detailed characterisation of anodic bonding
1.5 mm-thick glass [7]. process between glass and thin-film coated silicon sub-
It is known that glass cannot be anodically strates. Sens Actuator 86:103107
bonded to glass. However, research has found 7. Cozma A, Puers B (1995) Characterization of the elec-
trostatic bonding of silicon and Pyrex glass.
that this can be realized by depositing an interme-
J Micromech Microeng 5:98102
diate layer. The intermediate layer can be 8. Shih WP, Hui CY, Tien NC (2004) Collapse of
polysilicon, amorphous silicon, silicon nitride, or microchannels during anodic bonding: theory and
silicon carbide [9]. This has opened an easy route experiments. J Appl Phys 95(5):28002808
9. Berthold A, Nicola L, Sarro PM, Vellekoop MJ
to construct glass-based microfluidic systems
(2000) Glass-to-glass anodic bonding with standard
which are widely used for capillary electrophore- IC technology thin films as intermediate layers. Sens
sis. Other investigations into anodic bonding have Actuator A82(13):224228
A 74 Anthrax
Definition Synonyms
Antibody
Cross-References
Definition
AC Electro-Osmotic Flow
Electrokinetic Motion of Polarizable Particles
Antibodies are immune system-related proteins
Electroosmotic Flow (DC)
called immunoglobulins. An important compo-
Electrophoresis
nent of the immune system, antibodies are
Nonlinear Electrokinetic Phenomena
found in the blood of all vertebrates. The synthe-
Stotz-Wien Effect
sis, or manufacture, of antibodies is initiated
Unbalanced AC Field
when a foreign substance, referred to as an anti-
gen, enters the body. Lymphocyte cells respond
to the foreign substance by making an antibody
with a molecular arrangement that fits the shape
of molecules on the surface of the substance so
Applications of Induced-Charge
that the antibody combines with it. Common
Electrokinetic in Microfluidics
antigens are the protein components of bacteria
Yasaman Daghighi
and viruses.
Physical Sciences, Sunnybrook Research
Institute, Toronto, ON, Canada
Antigen Synonyms
Applications of Induced-Charge Electrokinetic in represent the external electric field directions, (b) geome-
Microfluidics, Fig. 1 (a) The computational domain of try and dimensions in a non-dimensional format, and (c)
the proposed ICEK microvalve with three microchannels the location of the four electrodes
and one suspended Janus particle in it. E1, and E2,
field. The dimension of this microvalve and elec- Applications of Induced-Charge Electrokinetic in
trolyte properties are listed in Table 1. To avoid Microfluidics, Table 1 Constants used for our numeri-
cal simulation
the gravity effect, the density of the fluid and the
heterogeneous particle are assumed to be equal. Parameters Values
Using the mathematical model described in the Dielectric constant, e 80
entry Induced charge electrokinetic motion of Permittivity of vacuum, 8.854 10 12
e0 (C V 1 m 1)
particles, the motion of the heterogeneous parti-
Viscosity, m (kg m1 s1) 0.9 10 3
cle in this microvalve can be numerically Density, r (kg m3) 998
simulated [5]. Particle diameter, Dp (mm) 20
Diffusion coefficient, Di (cm2 S1) 1.52 106
Induced-Charge Electrokinetic Micromixer Molar concentration at entrance 10
Mixing two or more fluid streams in A, CA (mol/m3)
microchannels without using microstirrer or Molar concentration at entrance 0
mechanical micromixers depends on their molec- B, CB (mol/m3)
ular diffusions; since, the nature of flow in
microchannels is laminar no convective term improve the mixing results can be achieved in
assists the mixing process. Thus, disturbing flow different ways such as the following: (i) using
to change the flow regime from laminar to turbu- complicated geometries and specific controlling
lent is a practical approach to develop a reliable systems, (ii) using mechanical microstirrers to
mixing process. Changing the flow pattern to agitate the flow, and (iii) implementing
Applications of Induced-Charge Electrokinetic in Microfluidics 77 A
Applications of Induced-
Charge Electrokinetic in
Microfluidics,
Fig. 2 Schematic diagram A
of the proposed micromixer
with two microchannels
and one suspended fully
conducting particle inside
the microchamber
electrokinetic methods for mixing. The induced- particle and the induced vortices by the resultant
charge electrokinetic concept and induced micro- applied DC electric field. The main function of
vortices can be employed to design an electric the electric field E2 is controlling the position of
micromixer with high performance [6, 7]. The the particle. Using the mathematical model
simplicity of such an ICEK mixing method has described in the entry, Induced charge electro-
attracted considerable attention in the fields of kinetic motion of particles, the motion of the
microfluidics and LOC [8, 9]. heterogeneous particle in this micromixer can
To improve the efficiency of the current be numerically simulated [10].
micromixer, a new nonmechanical micromixer The concentration field of different fluid
is introduced in this entry which works based streams is described by the concentration
on the ICEK phenomena. Consider one equation
microchamber which is connected to two
microchannels. The inlet channel is placed on @Ci
the left-hand side of the chamber and delivers U Ci Di 2 Ci i 1, . . . , n (1)
@t
two parallel streams of two different fluids into
the microchamber from entrances A and
B (Fig. 2). On the right-hand side of the where Ci is the concentration of the fluid stream
microchamber, there is a microchannel which is i and Di is its diffusion coefficient. The boundary
used as an outlet. A circular fully conducting conditions are
particle is placed in the middle of the
microchamber. The density of this particle is C Ci at inlet
(2a)
chosen to be exactly the same as the fluid which i, i 1, . . . , n entrances A and B
is filled in the micromixer. The electrolyte prop-
erties and dimension of this micromixer are listed
in Table 1. @Ci
0 at all the walls (2b)
Four electrodes are used to control this @n
t
0
micromixer. E1 is responsible for generating the
electroosmotic flow to transport the fluid in and and the initial concentration in the system is
out of the mixing chamber, in addition to induc-
ing the vortices. To achieve the mixing function,
it is necessary to control the position of the Ci jt0 0 through the microchannel (3)
A 78 Applications of Induced-Charge Electrokinetic in Microfluidics
Key Research Findings microvalve requires 3.7 s to switch the flow direc-
tion without any leakage. This operation period
ICEK Microvalve can be controlled by changing the applied electric
Applying positive voltage to electrode 1 and neg- field; e.g., when applying 400 V/cm, the
ative voltage to electrode 2, the electric field E1 is microvalve requires only 2.3 s to switch the
generated from left to right. As a result of the flow direction. This switching time will be
applied electric field, vortices are induced around increased to 7.82 s when the applied electric
the conducting hemisphere of the heterogeneous field is reducing to 100 V/cm.
particle (Fig. 3a) and push the particle to move The flow rate of the fluid that is pumped out of
toward the microchannel B (Fig. 3b). Once the this microvalve is adjustable. This adjustment is
heterogeneous particle reaches the entrance of possible by using different values for E*1 or E*2 in
microchannel B and blocks it (Fig. 3c), the elec- the valid range without affecting the blocking
trodes 1 and 5 will be simultaneously activated to position of the heterogeneous particle. The
produce the electric field E1 = 200 V/cm. This numerical simulations show that while the dom-
particle does not move as long as the electric field inant electric field is E1 = 200 V/cm, the addi-
E1 is applied. The goal of blocking the desired tional electric field, E*2, has to be between
microchannel without leakage in a short period of 29 V/cm and 46 V/cm in order to have a no
time is achieved (Fig. 3c). However, since the leakage microvalve. Similarly, applying the dom-
electric field direction is from left to right, it is inant DC electric field of E2 = 200 V/cm, the
not possible to direct the fluid to flow into the additional electric field, E*1, can vary from
microchannel C (Fig. 3c). To reach this goal, 49 V/cm to73 V/cm to pump the flow toward
a second DC electric field (perpendicular to microchannel B, while the microchannel C is
E1 = 200 V/cm) is applied, E*2 = 35 V/cm. This blocked by the heterogeneous particle.
second electric field is used to drive fluid flow
through the microchannel C. This is chosen suf- ICEK Micromixer
ficiently weaker than E1 so that it will not affect Consider a microchamber described in Fig. 2
the blocking of microchannel B (Fig. 3d). The which has no particle inside. Two parallel
heterogeneous particle tends to align itself with streams of fluids enter the chamber, once the
the direction of the resultant DC electric fields DC electric field E1 is applied. One stream is
(summation of E1 and E*2) but still blocks the a buffer solution carrying a sample species with
microchannel B. a sample concentration of CA = C0. The other
To change the flow direction, E1 will be turned stream is a pure buffer solution with zero sample
off, while E2 = 200 V/cm is applied using elec- concentration, CB = 0 (mol/m3). The only mixing
trodes 3 and 4. Figure 3eg show that the hetero- mechanism (to mix these two parallel laminar
geneous particle rotates to align itself with the flows) is the diffusion at the interface region
new applied DC electric field and moves toward along the micromixer. As demonstrated in
the microchannel C and eventually blocks Fig. 4a, at the exit of the chamber, the two fluids
it. Once microchannel C is blocked, the electric are poorly mixed. The resultant mixture is not
voltage is applied to electrode 6 simultaneously acceptable to be used in microfluidic devices.
to provide the same E2 = 200 V/cm, and keeps Releasing a fully conducting particle into this
the heterogeneous particle at the entrance of microchamber, the ICEK micromixer is pro-
microchannel C (Fig. 3h). By applying voltages duced. The induced vortices will be formed
to electrodes 1 and 2, a weak DC electric field, around the conducting surface once the electric
E*1 = 50 V/cm, is provided which manipulated field is applied. These four induced vortices are
fluid flow toward the microchannel B (Fig. 3i). circulating the fluid across the streamlines of the
The simulation study showed that under the two different fluid streams and boost the mixing
applied DC electric field of 200 V/cm, this ICEK process. The fully conducting particle moves
Applications of Induced-Charge Electrokinetic in Microfluidics 79 A
Applications of Induced- a b
Charge Electrokinetic in
Microfluidics,
Fig. 3 Sequences of A
E1=200V/cm E1=200V/cm
a microvalve which is
operated by induced-charge
electrokinetics using C N/C C N/C
a heterogeneous particle
inside it. The flow pattern
and vortices are shown by t1 = 0.0 s t2 = 1.1 s
E2=0 V/cm E2=0 V/cm
plotted streamlines. The
normalized vectors show c d
the direction of the fluid.
The diameter of the
heterogeneous particle and E1=200V/cm E1=200V/cm
the microchamber are
20 and 40 mm, respectively.
The height of the C N/C C N/C
microchamber is 40 mm.
The results are presented at
the xz plane crossing the t3 = 3.3 s t4 = 3.6 s
E2=0 V/cm E*2=50 V/cm
middle of the 3D
microvalve at different time e f
steps. The zeta potential on
the nonconducting
hemisphere of the E1=0V/cm E1=0V/cm
heterogeneous particle is
set to 50 mV, while the N/C N/C
C C
zeta potentials on the
nonconducting
microchannel and
microchamber walls are set t5 = 4.9 s E2=200 V/cm t6 = 5.7 s E2=200 V/cm
to 15 mV g h
E1*=50 V/cm
N/C
C
Applications of Induced-Charge Electrokinetic in time steps. (d) Comparison of the concentration distribu-
Microfluidics, Fig. 4 Flow pattern and concentration tion at the outlet of the micromixer at t=12s. E1 represents
distribution in the micromixer. The red lines inside the the horizontally applied electric field and is equal to
microchamber represent the streamlines of the flow field. 40V/cm. DC electric field (E1=40V/cm) is applied from
The color bar shows the concentration distribution. The left to right. C represent conductive material. Zeta
figure shows a micromixing chamber (a) without a particle potential of channel and chamber walls is 50 (mV)
inside, (b and c) with a conducting particle at different
toward the exit of the chamber driven by the conducting particle away from the right-side wall
applied electric field E1. However, the particle of the microchamber. At the same time, two
stops moving when it gets close to the right-hand vortices on the left-hand side of the fully
side wall of the microchamber. The motion of the conducting particle push the particle to the right.
rotating fluid (caused by vortices) is restricted by When these forces are balanced, the particle
the microchamber wall. Thus, the momentum of remains at an equilibrium position. Conse-
the fluid motion converts to pressure, which quently, the particle will not block the entrance
results in a repellent force between the wall and of the right microchannel. As shown in Fig. 4c the
the particle. This repellent force pushes the fully induced vortices around the fully conducting
Applications of Induced-Charge Electrokinetic in Microfluidics 81 A
particle significantly enhance the mixing process. symmetrical to the center line of the
The concentration profile at the exit of the microchamber all the time (Fig. 4b, c). Each of
microchamber is plotted in Fig. 4d for these two these induced vortices covers only one-quarter
investigated cases. These numerical simulations region of the microchamber, and they are located
illustrated that using a conducting particle inside in one stream only. In order to improve the
the micromixer enhances the mixing process. mixing result, it is desirable to have each induced
Under the applied only one DC electric field, vortex be located across the boundary of the two
E1, from left to right, the induced vortices are streams. Thus, such a vortex can circulate fluid
A 82 Arnoldi-Based Model Order Reduction
Cross-References Synonyms
Anisotropic Etching
Artificial Neural Networks
Plasma Etching
Reactive Ion Etching (RIE)
Synonyms
RIE Lag
ANN
Assay
Definition
Synonyms
An artificial neural network is an information
processing paradigm inspired by way the human Test
nervous system processes information. It consists
of an interconnected group of processing ele-
ments (neurons) which can exhibit a complex Definition
global behavior, determined by the network
topology (the scheme used to connect elements A method for the determination of the level of a
together), and the algorithm (the rules) employed given analyte in a given sample or set of samples.
to specify the values of the weights connecting
the nodes, the connections between the neurons.
Cross-References
Based on training rules (whereby the weights of
connections are adjusted on the basis of presented
Lab-on-a-Chip Devices for Chemical Analysis
patterns), ANNs can be trained to generate an
arbitrary number of outputs in response to an
arbitrary number of inputs.
Atomic Force Microscope (AFM)
Cross-References Definition
AFM Cross-References
Fabrication of Self-Assembled Catalytic
Nanostructures Piezoelectric Microdispenser