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Volume 17
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7 January 2017
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DOI: 10.1039/C9LC01112A
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ARTICLE

AHSWYE Microfabricated electrochemical sensing devices

748YIHURJ4NRGEK HF7B
Fumihiro Sassa,a Gokul Chandra Biswasb and Hiroaki Suzukic
Electrochemistry provides possibilities to realize smart microdevices of the next generation with high functionalities.

Lab on a Chip Accepted Manuscript

Received 00th January 20xx,
Accepted 00th January 20xx
DOI: 10.1039/x0xx00000x
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Electrodes, which constitute major components of electrochemical devices, can be formed by various microfabrication
techniques, and integration of the same (or different) components for that purpose is not difficult. Merging this technique
with microfluidics can further expand the areas of application of the resultant devices. To augment the development of
next generation devices, it will be beneficial to review recent technological trends in this field and clarify the directions
required for moving forward. Even when limiting the discussion to electrochemical microdevices, a variety of useful

www.rsc.org/ techniques should be considered. Therefore, in this review, we attempted to provide an overview of all relevant
techniques in this context in the hope that it can provide useful comprehensive information.

Introduction electrochemical principles, e.g., transistor-based devices.

Electrochemical sensing devices comprise a major group in
the field of bio/chemical sensing. There are many advantages
to using electrochemistry in sensing. First, a wealth of
techniques can realize highly sensitive detection with rapid
response times. Second, because the major components of
electrochemical devices are electrodes, miniaturization, batch
fabrication, multiplexing of the same components, and
integration with other components on a single chip can be
performed. Furthermore, electrochemical devices can be
operated using simple peripheral instruments with low power
consumption, and the instruments to operate the devices are
also simple and portable. Many successful devices have
already been commercialized.
The necessity of highly sensitive rapid detection of chemical
and biological analytes in samples of very small volumes is
increasing in applications such as point-of-care testing
(POCT).1-5 To handle solutions of very small volumes,
microfluidics provides a means to offer additional functions to
electrochemical sensing devices. Data obtained using such
devices can provide critical chemical/biological information
such as cell functions.
This review is written with the hope of providing a
comprehensive view of related techniques and devices that
could assist in the development of new next-generation micro-
electroanalytical devices. We will mainly focus on devices
reported within the last decade for liquid samples, along with
related techniques such as microfluidics. Depending on the
category, there are devices based on electrical rather than
a. Graduate
School of Information Science and Electrical Engineering, Kyushu
University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395.
b. School of Life Sciences, Shahjalal University of Science and Technology, Sylhet-
However, we limited the scope of the investigation to ensure
the range of discussion was not expanded unnecessarily and
the paper focuses on the main objective of reviewing
techniques and devices. In our discussion, we also mention
useful critical techniques originally developed for devices with
dimensions in the order of mm (or larger) that are not used for
microdevices.
In completing this review, our impression is that there are
really a wide variety of techniques for electrochemical sensing
devices. For specialists of such devices, there may be a
discovery of novel useful techniques. Also, the information
provided in this review will be useful to know the state-of-the-
art of this theme. We hope this review will contribute to the
development of new electrochemical sensing devices.
Basic components and fabrication
Basic setups of electrochemical devices.
We will start by briefly mentioning basic components,
setups, and instruments used in electrochemical devices. For
amperometry and voltammetry, a three-electrode system is
typically used, consisting of the working, reference, and
auxiliary electrodes. Two working electrodes are sometimes
used for purposes such as redox cycling. The three-electrode
system is operated by a potentiostat. A constant potential or
varying potential of a waveform is applied to the working
electrode with respect to the reference electrode and current
generated by a redox reaction is measured. No current flows
through the reference electrode and current flows through the
working and auxiliary electrodes. Coulometry is a variation of
amperometry. Here, the output is the charge, which is the
integral of current.

3114, Bangladesh.
c. Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1

Tennodai, Tsukuba, Ibaraki, 305-8573, Japan.

Electronic Supplementary Information (ESI) available: [details of any
supplementary information available should be included here].

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Fumihiro Sassa is currently an
Assistant Professor at the
department of electronics at
Kyushu University, Japan. He
achieved his Ph.D. on
microfluidics and electro-
chemical sensors in 2011 from
University of Tsukuba, Japan. His
research focus is on developing
components and systems of
Fumihiro Sassa micromechanical systems and
microfluidic systems employing
soft microfabrication techniques.
Gokul Chandra Biswas is
currently an Associate Professor
at the Genetic Engineering and
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Biotechnology Department at
Shahjalal University of Science
and Technology, Bangladesh. He
achieved his Ph.D. on Nano-
Science and Nano-Technology in
2017 from University of Tsukuba,
Japan. His research focus is on
Gokul Chandra Biswas the development of simple, low-
cost, and automatic biosensing
systems for bio-medical,
agricultural, food safety,
toxicological and environmental
analyses.
For potentiometry, a basic configuration uses two
electrodes: an indicator electrode to detect an analyte, and a
reference electrode. The potential of the former is measured
with respect to the latter using an electrometer. The potential
changes linearly with respect to the logarithm of the
concentration of the analyte, as expressed by the Nernst or
Nikolsky–Eisenman equations.
In impedimetry, the basic setup may be the three-electrode
system mentioned previously, and changes in impedance
around the working electrode are measured. However, in most
practical sensing devices, a pair of electrodes is often used
(including interdigitated electrodes made with the same
metal). The impedance between the electrodes is analyzed
using an impedance analyzer.
Thin- and t hick-film electrochemical microdevices, including
the three-electrode systems and microelectrodes, are already
available from many commercial sources. Accordingly,
numerous electrochemical microdevices are constructed using
these commercially available devices. We did not include these
devices in the following discussion to focus on the creation of
new devices rather than the progress in detection chemistry.
Materials and formation of electrodes
In electrochemical devices, the most critical components are the
electrodes. For the working electrodes, materials such as platinum,
2 | J. Name., 2012, 00, 1-3
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Hiroaki SuzukiDOI:received his B.E.
10.1039/C9LC01112A
and M.E. in applied physics and
his Ph.D. in bioelectronics and
biotechnology from the University
of Tokyo, Japan, in 1981, 1983,
and 1993, respectively. In 1983,
he joined Fujitsu Laboratories,
Ltd. as a researcher. In 1996, he
moved to University of Tsukuba as
an Associate Professor. Since
2004, he has been a full-time
Hiroaki Suzuki
professor at the Graduate School
Lab on a Chip Accepted Manuscript
of Pure and Applied Sciences,
University of Tsukuba. From
2016-2018, he was the president
of the Sensors and Micromachines
of the Institute of Electrical
Engineers of Japan (IEEJ).
Currently he is the president of the
Japan Society of Chemical
Sensors, a fellow of the Royal
Society of Chemistry, and a fellow
of IEEJ.
gold, and carbon are often used. These electrodes are usually
formed by thin- or thick-film techniques, including photolithography,
electron-beam lithography, focused ion beam (FIB) milling, and
screen-printing. Thin-film techniques are indispensable for creating
fine patterns (such as interdigitated electrodes used for redox
cycling) and promoting integration. On the other hand, they require
expensive facilities and cost for fabrication often poses a problem.
When a single sensor or a few sensors is/are sufficient for the
intended purpose and miniaturization is not strictly required,
screen-printing along with the use of plastic substrates will be more
cost-effective and a practical choice.
Materials other than those mentioned above have also been
used in attempts to improve performance. Boron-doped diamond
(BDD) features a wide potential window and low background
current.6 Microfabricated BDD electrodes have been used in the
detection of heavy metal ions,7 detection of dopamine,8 and
screening of NAD(P)-dependent oxidoreductases.9 Depending on
the application, it can be preferable to form an insulating layer with
diamond. Coplanar BDD electrodes of desired patterns insulated by
an insulating diamond can be formed by laser micromachining (Fig.
1A).10 An all-diamond structure can also be fabricated by patterning
layers of BDD and an insulating diamond to form a stacked-layer
structure.8
For the formation of carbon electrodes, a unique method,
pyrolysis of a photoresist, has been used 11 and carbon electrodes
thus fabricated have been used in some devices reported
recently.12-17 With this method, photoresist patterns are first
formed and then converted into carbonaceous materials at a high
temperature. Carbon electrodes of fine three-dimensional
structures can be formed by pyrolysis after patterning (Fig. 1B).12-14,
16, 17 Otherwise, polyimide can be used to form a patterned carbon
layer after resist patterning and dry etching.18 This technique is
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effective when relatively thick three-dimensional (3D) electrode
patterns are required.
As in the case of the carbon electrodes formed by pyrolysis,
three-dimensional structures are needed to form working
electrodes as well as structures such as reaction chambers and flow
channels. To this end, 3D-printing techniques have also been used
recently.19 The technique will be useful in forming flow channels, 3D
electrode structures, and other auxiliary structures such as
microneedles in the trial stage of development of new devices.
Electroplating is also useful to form nanostructures of metals such
as platinum, palladium, and gold additionally on patterned
electrodes to improve detection sensitivity.20-23
When an array or ensemble of electrodes of the nm scale needs
to form dual electrodes for redox cycling (Fig. 1D).28 The
View sensing
Article Online
area of 9 mm2 contained one billion nanopores DOI: 10.1039/C9LC01112A
with a lateral
distance of ~100 nm and pore radii of less than 20 nm.
For the formation of relatively large patterns, screen-printing is
widely used. However, it presents the issue of a non-negligible
amount of inks being wasted. In this regard, inkjet printing may
minimize the consumption of materials to be printed and therefore
the waste generated.29 With this method, not only electrodes but
also small patterns of biomolecules can be prepared. It also
facilitates multiple high-resolution deposition steps of molecules.
Although screen-printing is a practical technique to mass-
produce inexpensive disposable electrochemical devices, other
effective techniques have also been proposed. Song et al. used

Lab on a Chip Accepted Manuscript

to be formed over a relatively large area in the order of µm,
nanosphere lithography is convenient.24 In this method, a
monolayer of close-packed nanospheres is first deposited as a
template on a substrate with a layer consisting of electrodes and
insulators. The template is then used to form an insulating layer or
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template filtration to prepare electrodes (Fig. 1E).30 Electrode
patterns were cut on a polyvinyl chloride (PVC) tape using a laser
engraving machine, and the tape was attached to a porous
polyvinylidene fluoride (PVDF) membrane. Then, conductive
nanomaterials were deposited there by filtrating their diluted

a protecting layer on areas not protected by the nanospheres. After
removing the nanospheres, an array or ensemble of nanoelectrodes
is exposed (Fig. 1C). Depending on the structure, other techniques
such as electrochemical deposition or dry etching can also be
combined to deposit an additional metal layer or to expose
underlying metal layers further.24-27
Porous anodic aluminum oxide has also been employed to
fabricate high-density nanoelectrodes. The porous oxide features
high-density arrays of uniform and parallel pores. Hüske et al. used
the porous oxide directly formed on a platinum electrode pattern
solutions. The PVC tape was finally peeled off. With this method,
the composition and thickness of the deposited film can be
controlled precisely by simply changing the volume or composition
of the solutions. This technique is useful for the formation of
electrode patterns of carbon nanotubes (CNTs) and graphene.
Otherwise, roll-to-roll (R2R) printing (Fig. 1F) 31, 32 can be a powerful
technique to improve productivity remarkably.
Removing liquid from internal structure of sensors

Fig. 1 Methods to form electrode structures for electrochemical sensing devices. (A) All-diamond device with BDD microelectrodes fabricated by laser ablation,
BDD overgrowth, and polishing. Adapted with permission from ref. [10]. Copyright 2014, American Chemical Society. (B) Formation of carbon mesh electrodes by
pyrolysis of photoresist. Reprinted with permission from ref. [17]. Copyright 2018, Elsevier. (C) Nanopore electrode array formed by nanosphere lithography and
reactive ion etching. (i) Formation of a nanoelectrode array. (ii) SEM image of the nanoelectrode array. Adapted with permission from ref. [27]. Copyright 2016,
American Chemical Society. (D) SEM image of the cross section of nanoporous dual electrodes. Adapted with permission from ref. [28]. Copyright 2014, The
royal society of chemistry. (E) Formation of electrode patterns by filtration. (i, ii) Patterning of CNT electrodes by filtration. (iii-v) PVDF filter with a PVC tape
patterned by laser cutting. Adapted with permission from ref. [30]. Copyright 2018, American Chemical Society. (F) Patterning of electrodes by roll-to-roll
printing. Adapted with permission from ref. [32]. Copyright 2018, American Chemical Society.

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Depending on the device, liquids can be additionally used as
internal components of the sensor. Ion-selective electrodes and
reference electrodes are representative examples. The presence of
a liquid does not pose serious problems in macroscopic
conventional electrochemical devices. However, it is an obstacle
toward achieving miniaturization.
As ion-selective electrodes, the so-called coated wire type
electrodes were often used, in which an ion-selective membrane is
directly formed on an electrode.33 However, a problem posed by
this structure was the mismatch of charge carriers between the ion-
selective membrane and the electrode. In the ion-selective
membrane, the carriers are ions. On the other hand, electrons are
the carriers in electrodes. To resolve this issue, an intermediate
layer is usually used as a redox mediator.34 Such materials can be
made of conducting polymers such as poly(3,4-
ethylenedioxythiophene) (PEDOT), polypyrrole, and poly(3-
octylthiophene) (POT). Otherwise, the double-layer capacitance at
the membrane–electrode interface of the coated-wire type
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electrodes was intentionally increased to eliminate factors that lead
to undesirable drift of the electrode potential. Carbon materials
such as graphene, graphene oxide, and CNTs have been used for
this purpose.34 Advances toward improved solid-state ion-selective
electrodes have been described well in a recent review by Hu et
al.35 Among the microfabricated ion-selective electrodes reported
so far, coated-wire type electrodes feature frequently. Although
previous studies to realize solid-state ion-selective electrodes were
not necessarily directed toward miniaturization, the techniques will
be indispensable in realizing reliable microfabricated ion-selective
electrodes, and the use of these techniques should be considered
seriously.
Miniaturization and integration of reference electrodes
Although the reference electrode may be an uninteresting
component that does not usually attract attention, it is
indispensable and its performance directly influences that of
the entire electrochemical devices. The required stability and
precision depend on the mode of operation. In amperometry,
the potential of the working electrode is usually fixed at the
diffusion-limiting region at which the generated current does
not depend on electrode potential. In this case, the shift in the
reference electrode potential is not serious, and even
polarizable electrodes such as those made of platinum, gold,
and carbon can be suitable alternatives, as observed in the
case of numerous microfabricated amperometric devices. On
the other hand, in potentiometry, the shift in the reference
electrode potential directly affects the reading of the potential
of the indicator electrode and causes a serious error.
Therefore, currently, commercially available reference
electrodes (in the order of cm) are used in many miniaturized
potentiometric devices.
As mentioned previously, requirements pertaining to the stability
of the reference electrode potential are not so strict in
amperometric sensing. Therefore, the simple so-called pseudo
reference electrodes (that are in direct contact with an analyte
solution) are often used under limited conditions. The used
electrodes are not restricted to non-polarizable electrodes such as
Ag/AgCl, and even polarizable electrodes such as platinum, gold,
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and carbon (which are formed simultaneously with the working
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Article Online
auxiliary electrodes) are used. However, this DOI:approach
10.1039/C9LC01112A
does not
work for potentiometric sensors, and an additional structure to
maintain the concentration of primary potential-determining ions is
needed, as noted for commercialized liquid-junction electrodes.
To realize reliable miniature reference electrodes for
microfabricated electrochemical devices, a direct approach is to
miniaturize the conventional liquid-junction Ag/AgCl electrode with
an internal filling solution. The potential of the Ag/AgCl electrode
depends on the concentration of Cl− ions as expressed by the
Nernst equation. In addition, the potential difference generated at
the liquid junction can be a cause of measurement error. To avoid
issues related to this phenomenon, a concentrated potassium
Lab on a Chip Accepted Manuscript
chloride (KCl) solution is generally used to maintain the
concentration of internal Cl− ions at the same level and to ensure
sufficient efflux of internal KCl and minimize the liquid junction
potential. As the total volume of the internal solution decreases,
however, it becomes more challenging to meet these requirements.
However, except for cases wherein a very thin probe is used or the
available space is very limited—as in the case of chips of the sub-
mm order—the situation is not so dire, because the chip needs to
conform to a certain size to ensure easy handling. As the size of the
liquid-junction electrode increases, its incorporation in a device and
achieving satisfactory performance becomes easier.
Considering long-term storage, the presence of an internal
solution is problematic as in the case of the ion-selective
electrodes mentioned previously. It will be ideal if the
reference electrode functions in a completely dry state. This is
a long-standing issue and many groups have attempted to
develop the so-called solid-state reference electrodes.36, 37 To
this end, structures similar to those employed in solid-state
ion-selective electrodes have been attempted to be
fabricated.38 In providing a reliable reference potential, the
most difficult step will be to design the outermost part of the
reference electrode similar to a liquid junction that is in
contact with the solution. The interfacial potential difference
between this part and the analyte solution should not change
when it is in contact with the solution that contains various
ions of various concentrations. Reference electrodes that
provide a stable potential have been fabricated by employing a
hydrophobic ionic liquid as the outermost layer to resolve this
problem.39-42 However, another issue is the use of the liquid,
which is complicated to incorporate in microfabricated devices.
Although its use may not be difficult for relatively large
structures such as those found in paper-based devices,
breakthroughs are required to enable its use in more
miniaturized devices.
Merging electrochemical microsensors with
microfluidics
Amperometric and voltammetric devices
Independent electrochemical devices such as screen-printed
three-electrode systems and interdigitated electrodes for
impedance measurement or redox cycling are commercially
available. Accordingly, there are many reports of microsensors
using these commercialized products. The novelty of these devices
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lies mainly in the detection of specific analytes, the method of
detection, and applications. We will not deal with them here except
for cases that contain relevant techniques of note.
Some notable works have attempted to integrate sensing and
solution processing. Horak et al. fabricated an amperometric device
with functions including reagent delivery for the detection of
Epstein–Barr virus infection.43 Fragoso et al. fabricated an
amperometric device for cancer markers (Fig. 2A).44 Sixteen
working electrodes were placed in separate chambers along with
five reservoirs to store reagents and samples. The reservoirs were
interfaced by septa and the solutions were moved from the
reservoirs to the sensing region by applying pressure. Lee et al.
fabricated a device for amperometric detection of C-reactive
protein (CRP) (Fig. 2B).45 The chip consisted of electrodes screen-
printed on a polyethylene terephthalate (PET) film, flow channels,
and a cover layer. Complexes consisting of CRP and alkaline
phosphatase were formed by an antigen–antibody reaction and the
product was detected. Lee et al. fabricated a device for the
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voltammetric detection of four biomarkers including fibrinogen,
adiponectin, low-density lipoprotein, and 8-isoprostane.46 The
device had five chambers with three-electrode systems, one of
which was used as the reference. Transport of solutions was
controlled by pneumatic microvalves. Kimura et al. fabricated a
device for the early diagnosis of bovine subclinical mastitis.47
Neutrophils in the raw milk of mastitic cows were separated from
fat globules in a double-layer flow channel structure and were
finally trapped and concentrated in a working electrode consisting
of an array of micropillars with immobilized superoxide dismutase.
Superoxide anions secreted from neutrophils were detected, which
were used to estimate the number of neutrophils and the
seriousness of the disease. Kim et al. used porous membranes to
control flows in multiple flow channels in a structure fabricated by
stacking plastic layers (Fig. 2C).48 They conducted electrochemical
immunoassay for the creatine kinase-myocardial band test.
Although continuous flows of liquids are predominantly used in
microfluidic systems, plug (or droplet)-based microfluidics will be
more beneficial in creating devices with higher functionalities. First,
compared with continuous flows, the volume of each plug can be
reduced remarkably in such cases. Second, multiple plugs can be
processed in a single flow channel. Third, the mixing of components
can be very rapid (<1 s) after merging two plugs and moving the
merged plug in a flow channel. In many previous devices, plugs of
aqueous solutions were formed by injecting the solution into
another immiscible carrier liquid using a branched flow
channel structure, which could be used for various purposes.49,
50 However, a serious problem with the application of this
method in electrochemical sensing is that electrode surfaces
may be contaminated by the carrier liquid. This difficulty can
be alleviated by separating the plugs by air. Based on plug-
based microfluidics, Itoh et al. fabricated a device to measure
the K-value for the freshness of fish (K-value) (Fig. 2D).51
Solution plugs were formed, merged to mix components, and
stopped at two sensing sites using auxiliary structures in a
single flow channel. The sums of concentrations of ATP-related
compounds to calculate the K-value were measured
amperometrically using two enzyme electrodes in the flow
channel. Plug-based solution processing can be a good
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alternative for exchanging sample and reagent View solutions for
Article Online
affinity sensing. A row of plugs including DOI: 10.1039/C9LC01112A
washing plugs
separated by air can be stored in a single tube or flow channel
and can be delivered to the sensing site sequentially.52 Yang et
al. used this technique for amperometric detection of tumor
biomarkers.53
As mentioned previously, electrochemical techniques are
advantageous in many areas of bio/chemical sensing.
Nevertheless, depending on the analyte, optical
measurements may be more suitable. Accordingly, all-around
sensing devices can be realized in the form of a hybrid,
including sensors based on other detection principles. Ordeig
et al. reported a hybrid device with integrated flow channels,
Lab on a Chip Accepted Manuscript
an electrochemical cell, microlenses, and air mirrors for
amperometric and absorbance measurements.54
Electrochemical devices also provide a method of placing animal
or microbial cells directly on the chip and measuring the changes in
activity. Li et al. fabricated a device with a culture chamber and four
flow channels located downstream, for the amperometric detection
of reactive oxygen and nitrogen species secreted from a population
of macrophage.55 Ges et al. fabricated a device to monitor the
secretion catecholamine from adrenal chromaffin cells (Fig. 2E).56
An array of U-shaped structures was formed to trap the cells and
catecholamines were detected by amperometry. Measurement of
the respiratory activity of cells is required in many situations such as
drug screening applications. Weltin et al. fabricated a device with a
cell cultivation chamber and sensors for pH, oxygen, lactate, and
glucose to monitor the metabolism of human cancer cells.57
Extracellular acidification, respiration of the cells, glucose
consumption, and lactate production were measured using the
sensors. Marzioch et al. fabricated a sensing device for oxygen and
temperature to check the efficacy of photodynamic therapy.58
Oxygen consumption of T-47D breast cancer cells was measured
during the therapy. Tanabe et al. fabricated a microfluidic device
with a miniaturized Clark-type oxygen electrode to measure the
bactericidal activity of neutrophils, which is closely related to
mental stress.59 Neutrophils and E. coli cells were co-cultured, and
changes in the respiratory activity of E. coli were measured using
the oxygen electrode.
Three-electrode systems have also been used for amperometry
in lab-on-a-disc (LoD) devices. In LoD devices, centrifugal force
moves solutions from the inlet reservoirs near the center of
rotation to compartments in the periphery, for metering, mixing,
reaction, and detection. Nwankire et al. fabricated an LoD device
for the quantification of ovarian cancer cells.60 The device
contained functions including blood separation, cancer cell
extraction from the plasma, specific capture, and detection by
impedance measurements. Five different assays could be
performed in parallel. Sanger et al. fabricated an LoD device with
eight integrated filtration units for sample pre-treatment and three-
electrode systems for cell-free detection of p-coumaric acid
produced by E. coli. (Fig. 2F).61 Li et al. developed an LoD device
with electrochemical sensing and blood sample separation
functions.62 The device was used for the determination of glucose,
lactate, and uric acid.
A problem with LoD devices is determining how to establish
electrical contact between the components on the disk and external
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instruments. A simple method is to stop the rotation and conduct
measurements in a stationary state. However, this method causes
delay and may influence time-sensitive experiments. Ideally, it is
better to conduct measurements without stopping the rotation. To
solve this problem, slip-ring setups have been used.63, 64 Kim et al.
used this setup and integrated functions including the capture of
analyte molecules, a bead-based enzyme-linked immunosorbent
assay, and amperometric detection of C-reactive proteins.63
Rectangular chips with electrodes fabricated separately were
incorporated in the disc. Paraffin wax valves (containing iron oxide
View Article Online
nanoparticles used as heaters) were employed DOI: 10.1039/C9LC01112A
for flow control
through melting them by laser radiation. Alternatively, Bauer et al.
coupled a battery-powered bipotentiostat to a disc with electrodes
to send the signal wirelessly.65 Furthermore, LoDs reported recently
by some groups provide power wirelessly to drive an on-chip
potentiostat along with wireless signal transmission.66, 67
Many current microfluidic systems rely on external bulky
pumps and valve systems. To promote automatic solution

Lab on a Chip Accepted Manuscript

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Fig. 2 Electrochemical sensing devices coupled with microfluidic functions. (A) Microfluidic device for electrochemical detection of breast cancer markers.
Adapted with permission from ref. [44]. Copyright 2016, The royal society of chemistry. (B) Device for amperometric detection of CRP. Adapted with permission
from ref. [45]. Copyright 2012, Springer. (C) Multilayered polymeric cartridge. (i) Exploded view, (ii) Cross-section of stacked layers. (iii) Top view of cartridge.
Reprinted with permission from ref. [48]. Copyright 2009, Elsevier. (D) Device to determine freshness of fish (I, ii)) PDMS substrate with flow channel structures
and glass substrate with electrodes. (ii) Complete device. (iii) Magnified view of sensing sites with bypass flow channels. Adapted with permission from ref. [51].
Copyright 2013, American Chemical Society. (E) Microfluidic platform for cell trapping and monitoring of cellular secretion. (i, ii, iii) Chromaffin cells in the traps,
(iv) Visualization of a perfusion flow using food dye. (i) Platinum electrodes. (ii) Cell trap. (iii) Chromaffin cells in cell trap. (iv) Flows disrupted by the clumps. (v)
Clumps of cells stuck between the traps. Reprinted with permission from ref. [56]. Copyright 2013, The royal society of chemistry. (F) Photograph of
electrochemical LoD device and magnified view of a unit containing chambers for filtration and detection for screening genetically modified E. coli cells. Reprinted
with permission from ref. [61]. Copyright 2017, Elsevier. (G) EWOD device with integrated potassium ion-selective electrodes. Reprinted with permission from ref.
[70]. Copyright 2018, Elsevier.

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transport, electrowetting is useful. In electrowetting on
dielectric (EWOD), wettability of a hydrophobic dielectric layer
formed on an array of electrodes separated with narrow gaps
is changed by the application of a voltage, and independent
droplets are moved sequentially. Owing to this feature, the
technique is often called digital microfluidics. An attractive
feature of EWOD devices is not only the transport of droplets
but also the capability of processing multiple solutions.
Programmable manipulation enables procedures such as
metering, transport, separation, and merging of droplets. In
the initial stage, electrodes used for sensing were formed
within the array of driving electrodes for EWOD on the same
bottom plate. Some groups integrated a three-electrode
system in such devices.68, 69 Farzbod et al. integrated an array
of potassium ion-selective electrodes (ISEs) at the bottom
plate (Fig. 2G).70 A problem with this layout is that the
electrodes are crowded and the electrodes for sensing must be
exposed without the hydrophobic dielectric layer, making
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fabrication more complicated. To overcome this problem,
sensing electrodes were subsequently formed on the top
ground plate.71-73 Shamsi et al. used such an EWOD device and
magnetic beads conjugated with primary antibodies to detect
the thyroid stimulating hormone.72 Rackus et al. formed
nanostructured gold working electrodes along with reference
and auxiliary electrodes to detect the Rubella virus by
electrochemical sandwich enzyme-linked immunosorbent
assay (ELISA).73
Potentiometric devices
The ion-sensitive field-effect transistor (ISFET) is a representative
electrochemical microsensing device. Although many reports
related to the ISFET were published until the last decade, they have
been decreasing in number since that time. Since the first report of
the ISFET in the 1970s, the technology has been established well
and faced challenges in various possible applications.74 Alternatively,
different field-effect transistor (FET) devices based on materials
such as carbon nanotubes or graphene have been actively studied.
It is a matter of controversy as to whether the detection principles
are electrical or electrochemical. Therefore, the new FET sensors
are not focused on here. Those who are interested in these devices
should refer to related reviews published recently.75-77
Regarding devices based on ISFETs, Yamada et al. fabricated
a microfluidic device for pH measurements.78 Two ISFETs and
an Ag/AgCl pseudo reference electrode were used for
differential measurements. Sharma et al. fabricated a
microfluidic chip with ISFETs for pH and potassium ions.79
Matsuura et al. fabricated a microfluidic device with an ISFET
to detect biofilm formation by measuring pH changes in the
culture medium.80 Andrianova et al. reported a microfluidic
device for the detection of organophosphorus pesticides. A
hydrolytic enzyme was used and pH changes were detected
using an ISFET.81 As an interesting application of the ISFET,
Kuznetsov et al. proposed a bioelectronic nose (Fig. 3A).82 An
ISFET with an immobilized aptamer for vanillin was immersed
in a solution in a flow channel and was separated from the gas
phase with a hydrophobic membrane with many holes that
acted as a gas-to-liquid extraction interface. The structure was
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formed by a sacrificial aluminum etching technique. The
View device
Article Online
selectively detected vanillin vapor. DOI: 10.1039/C9LC01112A
Lin et al. used an electrolyte–insulator–semiconductor structure
for the detection of glucose, urea, and creatinine.83 Alginate
microbeads containing magnetic beads and an enzyme were
localized using a step-like obstacle structure in a flow channel and a
magnetic field. Upon the injection of an analyte solution, enzymatic
reactions caused pH changes, which were measured.
Contrary to the ISFETs, ion-selective electrodes have been
used in various devices, particularly in paper-based and
wearable devices. In many of these applications, sensing areas
in the order of mm have been used and the electrode
potential could be easily measured. Details will be mentioned
Lab on a Chip Accepted Manuscript
later in corresponding sections.
Impedimetric devices
Wang et al. fabricated a device for the detection of
thrombin.84 Gold interdigitated electrodes were formed in a
polydimethylsiloxane (PDMS) flow cell. Magnetic beads
modified with aptamers were used to capture, separate, and
concentrate the target protein. Daprà et al. reported an all-
polymer electrochemical device using a conductive polymer
bilayer to form electrodes.85 The electrode was functionalized
with aptamer probes to detect ampicillin and kanamycin A by
electrochemical impedance spectroscopy(EIS). Ali et al.
fabricated a microfluidic device for the detection of low-
density lipoprotein using bovine serum albumin and
antiapolipoprotein B-functionalized carbon nanotube–nickel
oxide nanocomposite.86. Chiriacò et al. fabricated a device to
detect autoantibodies towards phosphorylated a-enolase for
the diagnosis of pancreatic ductal adenocarcinoma. The device
had two PDMS reaction chambers, each of which had four
couples of interdigitated electrodes with immobilized peptides
corresponding to a portion of the enzyme.87 Lakey et al.
fabricated a microfluidic device with five sets of gold
interdigitated electrodes for impedance measurements to
detect a biomarker (D-dimer) for the diagnosis of deep vein
thrombosis and pulmonary embolism.88 Anti-D-dimer antibody
was immobilized at the individual sensing sites following the
electrochemical patterning of polypyrrole.
As well as molecules, the detection, number counting, and
activity monitoring of animal or bacterial cells have also been
investigated using such techniques. Paredes et al. used an
impedimetric device to monitor the growth of a bacterial
biofilm.89 Etayash et al. conducted real-time, label-free
detection of Gram-positive bacteria (L. monocytogenes) that
acted as foodborne pathogens using antimicrobial peptides as
probes.90 Manczak et al. detected monocytes using
immobilized antibodies (Fig. 3B).91 An et al. used a benzoboric
acid-modified gold-plated polymeric substrate with a regular
3D microarray to detect rare circulating tumor cells (CTCs)
efficiently.92 The number of CTCs was measured by EIS and the
captured CTCs could be detached by a reversible reaction
between benzoboric acid on the 3D microarray and sialic acid
on CTCs. In addition to the orthodox methods, some
interesting methods were also used. Damhorst et al. coupled
Ion-encapsulating liposomes functionalized with antibodies
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with impedance spectroscopy (Fig. 3C).93 The analyte virus was
sandwiched between antibodies on the electrodes and
liposomes. The ions in the liposomes were released by heating.
Detection of human immunodeficiency virus (HIV) was
attempted using the device.
In the detection of DNA by label-free impedance spectroscopy,
hybridization in a stationary solution takes time. Wu et al.
accelerated hybridization by stirring solutions by an AC
electroosmosis (ACEO) flow (Fig. 3D).94 When animal or bacterial
cells are analyzed by impedimetry in microwells or microfluidic
channels, an asymmetric electrode configuration can be easily
established in the structure. This enables concentration of cells by
dielectrophoresis.95-99 When objects such as cells View are
Article more
Online
polarizable than the medium, they are draggedDOI:to
10.1039/C9LC01112A
a higher electric
field region around the smaller detecting electrodes, which is called
positive dielectrophoresis (pDEP). By contrast, when the medium is
more polarizable than the objects, they are repelled from the
higher electric field region, which is called negative
dielectrophoresis (nDEP).100 Mansoorifar et al. concentrated and
confined PC-3 prostate cancer cells within microwells by pDEP for
impedance measurements. 100, 101 The cells were treated with the
drug Enzalutamide, and dying cells exhibited different spectra
compared to those of healthy cells. Depending on the scenario, the
cells were released from the electrode and were used for other

Lab on a Chip Accepted Manuscript

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Fig. 3 Potentiometric and impedimetric devices. (A) Bioelectronic nose created with a field-effect transistor with an immobilized aptamer. Reprinted with
permission from ref. [82]. Copyright 2019, Elsevier. (B) Device used for on-chip impedimetric immuno-detection of subpopulations of cells. Reprinted with
permission from ref. [91 ]. Copyright 2016, Elsevier. (C) Impedimetric sensing of virus that employ the release of encapsulated ions from tagged liposomes.
Adapted with permission from ref. [93]. Copyright 2013, Springer. (D) Vortices induced over electrodes by AC electroosmosis for label-free impedimetric DNA
sensing. Reprinted with permission from ref. [94]. Copyright 2013, Elsevier. (E) Concentration of bacteria by negative and positive dielectrophoresis for
impedance measurement. Reprinted with permission from ref. [104]. Copyright 2013, Elsevier. (F) Steps for seeding cells into multiple chambers. (i) Flow channel
1 is filled with the solution, which do not enter the chambers. (ii) Each chamber is filled with the solution by having the solution withdrawn by applying a negative
pressure from port 2. (iii) The solution in flow channel 1 is removed by applying a pressure. Reproduced with permission from ref. [107]. Copyright 2016, The
royal society of chemistry. (G) Microfluidic device with a solution-processing function for label-free impedimetric detection of DNA hybridization analysis. (i) Chip
with electrodes. (ii) Assembled device with assay channels (green) and horizontal (blue) and vertical (red) channels for valve control. (iii) Layout of flow channels
in the stacked layers. Inset shows actuation of the valve. Reprinted with permission from ref. [108]. Copyright 2015, Elsevier. (H) Integrated automated
multiorgan-on-a-chip and sensing platform consisting of modular components, including electrochemical biosensors. Adapted with permission from ref. [117].
Copyright 2017, National Academy of Sciences.

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purposes. Nakano et al. collected bacteria on microelectrodes by
pDEP, and electropermeabilization was used to disrupt cell
membranes by applying a pulse-like rectangular wave. Intercellular
ions released by cell disruption were detected.102 Dastider et al.
used two sets of gold interdigitated electrodes in a flow channel to
detect E. coli O157:H7.103 A set of electrodes focused and
concentrated E. coli by pDEP to direct it to other sets of electrodes
with E. coli-specific antibodies used for detection. Hamada et al.
used both pDPE and nDEP (Fig. 3E).104 Bacteria introduced into the
device were first repelled by nDEP force exerted by the first
electrode and were captured by pDEP on the second electrode.
Chuang et al. used multi-layer electrodes to trap and release cells
from an array of microcavities and to conduct impedance
measurements.105 The device had an ITO top electrode, a middle
electrode on the top surface of the microcavity structure, and
electrodes at the bottom of the microcavities. By changing the
combination of electrodes, cells were trapped/released individually
in the microcavities by nDEP for impedance measurement of the
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bladder cancer cell. After the measurement, the trapped cells were
released by pDEP.
Devices have also been fabricated aiming toward multiplexed
detection. Onishi et al. fabricated an impedimetric device for on-
site determination of the minimum inhibitory concentration of
antibiotics.106 They formed an array of eight chambers for
impedance measurements to accommodate antibodies of different
concentrations (Fig. 3F).107 A sample solution containing bacteria
was introduced into the chambers and the solutions in each
chamber were separated by removing unnecessary solutions with
the help of surface tension. The growth of the bacteria in each
chamber was monitored by impedance measurements. Ben-Yoav et
al. formed 3 × 3 individually addressable reaction chambers for the
detection of DNA hybridization by EIS (Fig. 3G).108 Dual-layer
programmable valves were formed to control solution transport in
flow channels formed of rows and columns. The valves closed the
flow channels by bending their PDMS membrane downward. The
valves allowed the functionalization of multiple working electrodes
with probe DNAs without cross-contamination or requiring parallel
incubation or measurements with multiple samples.
Application to Organs-on-Chips
Presently, 3-D human tissue/organ models known as organs-on-
chips, are being developed as platforms for personalized medicine
and drug screening.109-112 These organs-on-chips facilitate the rapid,
inexpensive, and effective investigation of in vivo drug efficacy and
toxicity, and it is expected that their application will solve the
ethical problems associated with the use of animals. In this study,
the in situ measurement of the concentration of indicator
molecules secreted by cells was critical in obtaining the information
regarding the viability and metabolic activity of organs, and will also
be beneficial in the analysis of multi-organ interactions. Therefore,
integrating a sensing function in organs-on-chips is extremely
important.
Moya et al. integrated three inkjet-printed amperometric
dissolved oxygen sensors in a liver-on-a-chip,113 and primary human
and rat hepatocytes were cultured. Their oxygen consumption was
evaluated, and the concentration distribution of the dissolved
oxygen was determined. Bavli et al. developed a liver-on-a-chip
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with a sensing module for the measurement of oxygen, glucose,
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and lactate concentrations.114 The module DOI: measured
10.1039/C9LC01112A
oxygen
concentration via phosphorescence quenching by employing
microparticles loaded with a ruthenium-based dye. Additionally, it
measured glucose and lactate concentrations amperometrically.
Shin et al. identified a cardiac injury biomarker, which was
essentially the creatine kinase-MB in a heart-on-a-chip; this was
achieved by using an aptamer-based impedimetric
immunosensor.115 Variations in the concentration of the dose-
dependent creatine kinase-MB were observed, which were caused
due to the cardiac damage upon exposure to doxorubicin.
Moreover, to continually monitor the secretion of human albumin
and glutathione-S-transferase-alpha after administering
Lab on a Chip Accepted Manuscript
acetaminophen, Shin et al. integrated impedimetric immunosensors
in a human liver-on-a-chip.116 The immunosensors could be
regenerated repeatedly by using a specially designed microfluidic
module. In a modular organ-on-a-chip, Zhang et al. used
electrochemical protein biomarker sensors as well as optical pH, O2,
and temperature sensors.117 The electrode surfaces were
functionalized with antibodies for the biomarkers, and were
regenerated for repeated measurements. Drug-induced organ
toxicity was examined using dual-organ human liver-and-heart-on-
a-chip and liver-cancer-and-heart-on-a-chip model systems (Fig. 3H).
Some reviews regarding the integration of bio/chemical sensors
into organs-on-chips have been reported.118, 119 However, only a
few studies on the in situ monitoring of drug-induced organ toxicity
have been reported that employ organs-on-chips, and studies on
the integration of sensing functions into these organs-on-chips are
still at the initial stages. Thus, studies on the incorporation of
microfabricated electrochemical sensors in devices are required;
further, an enhancement in the monitoring performance, including
precision and multiplexed detection improvement, is highly
necessitated.
Paper-based devices
Compared with that of other electrochemical devices,
development of paper-based devices has been extensively
investigated over the last decade. In contrast to the
complementary metal oxide semiconductor (CMOS) devices
mentioned later, the paper-based devices are ultimately
simple, which is really advantageous in realizing inexpensive,
portable, and disposable devices. Since the first report,120
many electrochemical paper-based devices and reviews have
been reported.121-129
As long as fabrication is concerned, many devices used existing
techniques such as screen-printing with the only exception that the
substrates were paper. However, some unique techniques specific
to paper have also been proposed. Ink-jet printing is particularly
suitable for paper and was used to draw electrode patterns.130-132
Santhiago et al. also used a XEROX printer to print patterns of
carbon electrodes.133 Along with wax printing already used for
paper to form structures such as flow channels, this is very efficient
in mass-producing devices with electrodes. Otherwise, a pencil or
pen is used to draw patterns of electrodes, hydrophobic barriers,
and biomolecules.134-139 The method lets users design their original
electrode patterns themselves. Soum et al. used a digital plotter to
automate the drawing.140 Alternatively, carbon electrodes were
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formed directly on paper boards by pyrolysis using laser light (Fig.
4A). The carbon composed of graphene sheets with an enhanced
active/geometric area ratio and exhibited high conductivity.141 Zang
et al. also formed conductive molybdenum carbide–graphene
composite electrodes by laser-powered conversion from fibrous
paper soaked with gelatin-mediated inks containing molybdenum
ions.142 As an essentially different approach, Fosdick et al. proposed
the use of metal wires as electrodes.143 The electrodes can be
incorporated into the device structure by simply folding paper. The
wires can be cleaned and modified with molecules prior to
assembly.
Another specific feature of the paper is that it can be folded and
stacked easily. The technique that accompanies the folding of paper
is often called the “origami” technique. The technique has been
used to form not only device structures 143-146 but also to simplify
analytical procedures and realize user-friendly devices. Unlike
previously mentioned microfluidic devices that mostly rely on
pumps and valves, the transport of solutions in paper-based devices
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depends on the wicking of solutions, which makes the sequential
procedure that handles multiple solutions difficult. The origami
procedure is effective in circumventing this difficulty.147-149 Fig. 4B
shows an example of such a device reported by Ding et al. for the
potentiometric detection of organophosphate pesticides.149
They separated zones for an enzyme, an enzyme substrate,
and a sample solution on different tabs. When the device was
used, the tabs were folded following a predetermined sequence.
The technique was also used for the detection of pesticides.150 A
modified design was proposed by Yakoh et al. (Fig. 4C).151 Their
device consisted of folding paper and a movable reagent-
stored pad. When in use, the device is assembled, a sample is
spotted, and a carrier buffer is loaded in its reservoir. The
reagent-stored pad is then pulled to introduce reagents or
switch off fluid flow. In the former case, the buffer elutes the
reagent from the reagent-stored pad to the channel with
electrodes of the origami folding paper. The same steps can be
repeated for the other reagents. Finally, amperometric and
voltammetric measurements are conducted.
Wearable electrochemical devices
Wearable devices have been a hot topic over the last several
years. Their application is mainly directed toward personalized
health, fitness, and conditioning of athletes. Numerous papers
and many reviews have already been published on them, even
on those that involve electrochemical components.38, 152-163
Therefore, in this section, we will briefly introduce this class of
devices, focusing on the effort to realize properties that wearable
electrochemical devices offer.
Designing devices that meet inherent requirements
A challenging problem with wearable devices is that the device is
often subjected to mechanical deformations such as stretching and
twisting. In these situations, strain-enduring electrodes are
required. Bandodkar et al. developed inks to form stretchable
electrodes by screen-printing.164, 165 The inks employed CNTs and
elastomeric polyurethane as a binder.165
The electrodes printed using these inks with serpentine patterns
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withstood strains up to 500% with a negligible influence onOnline
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electrochemical performance. Lee at al. DOI: used10.1039/C9LC01112A
chemical vapor
deposition (CVD)-synthesized graphene doped with gold formed on
serpentine gold electrode patterns.166 The electrode showed
improved electrochemical activity over bare graphene.
In relation to the abovementioned problem, consistency of
wearability and electrical contact is another critical point to be
considered when using wearable devices. The electrodes used for
sensing must be flexible to conform to the non-planar skin surface.
To this end, electrodes that take the form of tattoos are very useful
(Fig. 5A).167 Jia et al. fabricated a temporary-transfer tattoo
amperometric enzyme biosensor for monitoring of lactate levels in
human perspiration (Fig. 5B).168 The technique was also used in
Lab on a Chip Accepted Manuscript
devices for the monitoring of epidermal pH levels,169 trace metals in
human perspiration,170 and ammonium ions in sweat.171
Depending on the application, there are some additional
requirements for the substrates. For example, when applied to
contact lenses to analyze the components of tears,172-174 the
substrate (and preferably electrodes) must be transparent. The
substrate must also fit the shape of the eyeball. In other words, the
electrodes must be formed on a substrate that is not flat. To realize
this, a method used in previous studies involved forming the
electrodes on a flat substrate and deforming it after that.
Furthermore, biocompatibility of materials in contact with the body
part is necessary, which entails stricter requirements when the
devices are used on the eye and in the mouth.
Considering wearability, it will be more preferable if the sensing
devices are formed on cloths or underwear. To this end, attempts
have been made to form electrodes on textiles. Choudhary et al.
used the textile weaving technique to form wearable devices.175 To
make electrodes, silk yarns were coated with conducting inks and
reagents before being woven. To restrict solutions to limited areas,
hydrophilic and hydrophobic yarns were also formed appropriately.
Liu et al. fabricated an embroidered electrochemical sensor (Fig.
5C).176 Electrodes were made from conductive threads and were
embroidered onto wearable fabrics.
Using hydrophilic patches to efficiently sample sweat
When sweat is used for analysis, sampling is considered to be
straightforward if perspiration is induced through exercise and/or
temperature elevation. However, sampling sweat under natural
perspiration conditions results in issues associated with the
transportation of analytes to the sensor surface. Therefore, to
facilitate efficient sampling following natural perspiration, a
hydrogel patch was formed on the electrodes that were meant for
amperometric detection.177, 178 Alternatively, perspiration can be
intentionally and effectively induced through the transdermal
delivery of a stimulating reagent such as pilocarpine to sweat
glands; this was achieved via the iontophoresis process, which
induces the flow of ions and molecules owing to the application of a
current across the skin. To deliver pilocarpine via iontophoresis and
potentiometrically measure chloride ion concentration, Gonzalo-
Ruiz et al. fabricated a device with screen-printed electrodes and a
hydrogel.179 Kim et al. integrated tattoo-like electrodes into
hydrogels for the iontophoretic delivery of pilocarpine and the
amperometric detection of ethanol alongside flexible wireless
electronics,180 whereas Emaminejad et al. integrated a hydrogel
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Fig. 4 Techniques used for paper-based electrochemical sensing devices. (A) Formation of carbon electrodes by laser pyrolysis and cutting. Adapted
with permission from ref. [141]. Copyright 2017, Wiley-VCH Verlag GmbH & Co. KGaA. (B) Folding sequence for a paper-based 3D origami
potentiometric device. Adapted with permission from ref. [149]. Copyright 2016, Wiley-VCH Verlag GmbH & Co. KGaA. (C) Paper-based device that
controls solution transport by folding and sliding of paper for electrode modification and voltammetric determination. Adapted with permission from
ref. [151]. Copyright 2019, American Chemical Society.

Lab on a Chip Accepted Manuscript

layer as well as Na+, Cl−, and glucose sensors alongside
iontophoresis electrodes.181 Acetylcholine, methacholine, and
pilocarpine were loaded in the hydrogel; further, iontophoresis was
performed wirelessly, while perspiration was induced at periodic
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Contrary to iontophoresis, reverse iontophoresis renders the
non-invasive sampling of interstitial fluid (ISF) possible. In this
regard, Kim et al. developed a device for the simultaneous and
independent sampling and analysis of ISF glucose and sweat-

time intervals without causing skin damage.
alcohol.182 This device employs the reverse iontophoretic extraction
of ISF and the iontophoretic delivery of pilocarpine into the skin. For
iontophoresis, the device used tattoo-like electrodes, and for
wireless real-time measurements, screen-printed electrodes
formed on a flexible substrate alongside readout circuits were used.

Use of minimally invasive microneedles

Real-time monitoring of metabolites in physiological fluids is
necessary in fields such as diabetes care and sports medicine. In the
development of wearable devices, body fluids such as sweat, saliva,
and tears are often used. However, when more accurate reliable
data are required, direct analysis of whole blood will be more
preferable. In conducting this, a problem is that sampling through a
needle accompanies pain. As a solution, the application of
minimally invasive microneedles has been tried mainly for the
analysis of ISF.
Coupling the microneedles with electrochemical detection using
microdevices can involve two methods. In one method,
electrochemical detection is conducted in a microfluidic system
with hollow microneedles formed vertically at the end of the flow
channel structure. Analyte solutions are withdrawn into the interior
of the device and detected electrochemically. In the other method,
electrodes are formed with microneedles to fabricate on-body
patch-type devices. The electrodes are directly stuck onto the skin
and electrochemical measurements are conducted.
As a device based on the former, Miller et al. formed a
microneedle on a sensing device for K+ ions by two-photon
polymerization.183 The same group also demonstrated the
amperometric detection of myoglobin and troponin using a similar
device with the microneedle.184 Jina et al. reported a device for
continuous glucose monitoring (Fig. 5D).185 The device had an array
of 200 silicon hollow microneedles with a length of ~300 μm and a
50 μm × 50 μm lumen. A screen-printed 3-electrode system
containing a working electrode with immobilized glucose oxidase
was formed in a sensing chamber for the amperometric detection
of glucose. The sensing chamber was initially filled with a buffer
solution. Glucose diffused from the epidermal layer of the skin
through the microneedle array into the buffer solution passively

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Fig. 5 Techniques used for electrochemical wearable devices. (A) Formation of temporary transfer tattoo. (a) The electrode (red) is patterned on the paper
(orange). (b) The adhesive sheet (blue) with protective coating (maroon) is applied to the paper with the electrodes. (c(i)) The protective sheet is removed. (c(ii))
The tattoo is flipped and applied to the skin (green). (c(iii)) The release agent-coated paper is removed and the electrodes are exposed to the wearer's external
environment. In the case that physiological monitoring is desired, the procedure d(i) - d(iii) is followed. (d(i)) The paper is removed. (d(ii)) The tattoo electrode
patterns are applied to the skin. (c(iii)) The protective coating is removed. Adapted with permission from ref. [167]. Copyright 2012, The royal society of
chemistry. (B) Three-electrode “NE” tattoo for the determination of lactate. Adapted with permission from ref. [168]. Copyright 2013, American Chemical Society.
(C) Embroidered Three-electrode systems formed with carbon-coated conductive thread (for W.E. and A.E.) and Ag/AgCl-coated thread (for R.E.). (i-iii) Pictures of
embroidered electrodes on a textile chip, cotton gauze, and cotton t-shirt. (iv) Magnified image of the part indicated in (iii) on the t-shirt. Adapted with
permission from ref. [176]. Copyright 2016, The royal society of chemistry. (D) Amperometric sensing device with an array of microneedles. (i) SEM image of
device. (ii) Magnified SEM image of microneedles. Adapted with permission from ref. [185]. Copyright 2011, Wiley-VCH Verlag GmbH & Co. KGaA. (E)
Mouthguard biosensor with wireless amperometric circuit board. Reprinted with permission from ref. [191]. Copyright 2015, Elsevier. (F) Mouthguard biosensor
for saliva glucose detection. Reprinted with permission from ref. [193]. Copyright 2015, Elsevier. (G) Active contact lens device for wireless health monitoring.
Adapted with permission from ref. [194]. ©2012, IEEE. (H) Glove biosensor with a sensing finger, immobilized organophosphorus hydrolase, and the
collector/sampling finger. The electrodes are connected via a ring bandage to a portable potentiostat on the back of hand with wireless Bluetooth communication
to a smartphone. Adapted with permission from ref. [196]. Copyright 2017, American Chemical Society.

and was detected on the working electrode. The device could
provide clinically accurate results over 72 h.
For direct amperometric measurements, working electrode
structures with microneedles of various designs have been
proposed. Windmiller et al. proposed a working electrode structure
constructed with an array of platinum-coated solid microneedles
covered with another array of hollow microneedles that surround
the solid microneedle.186 The microrecesses formed between the
two types of microneedles were used to immobilize enzymes in a
poly(o-phenylenediamine) (PPD) layer by electropolymerization.
Detection of glutamate and glucose was demonstrated using this
working electrode structure. The PPD layer rejected electroactive
interferents effectively and the microrecesses protect the enzymes
upon insertion into the skin. Caliò et al. formed a working electrode
structure with polymeric microneedles with an immobilized enzyme
formed on a flexible substrate.187 The microneedles were formed
lactate oxidase. The same group also fabricated a device with two
working electrodes for the simultaneous monitoring of lactate and
glucose concentrations.189 The performance of the devices was
assessed in artificial ISF and in human serum. Microneedles were
also combined with wearable devices to deliver drugs
transcutaneously. Lee et al. fabricated a patch consisting of a
heater, temperature, humidity, glucose, and pH sensors along with
polymeric microneedles.166 The device measured glucose
concentration in sweat and the microneedles were thermally
activated to release a drug at high glucose concentration.
Wireless signal transmission and power transfer
Considering the user-friendliness of wearable devices, wireless
device operation and data transmission will be a must as
demonstrated by many recently reported devices. 166, 173, 180-182, 190-
192 In addition, wireless energy transfer will be another option

on a gold electrode by photolithography using a photocurable
mixture containing an enzyme. They demonstrated the detection of
glucose and lactic acid. Bollella et al. reported a working electrode
structure for the sensing of lactate in dermal ISF.188 Electrodes were
formed on a structure of 64 polycarbonate microneedles. Gold and
multiwalled carbon nanotubes were electrodeposited followed by
the electropolymerization of methylene blue and immobilization of
to be considered. Bandodkar et al. fabricated a Na+ ion-selective
tattoo electrode paired with a wireless transceiver for the
monitoring of sweat Na+ ion concentration on the human
epidermis.190 Saliva and tears were also used for the analysis using
wearable devices. Kim et al. fabricated a mouthguard that can
monitor the salivary uric acid level (Fig. 5E).191 Screen-printed
electrodes with immobilized enzyme along with compact electronic

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circuits including a potentiostat, microcontroller, and a Bluetooth
transceiver were incorporated in the mouthguard. Arakawa et al.
fabricated a device used in the human oral cavity for the monitoring
of saliva glucose (Fig. 5F).193 The device consisted of a platinum
electrode with immobilized glucose oxidase along with a Ag/AgCl
electrode on a mouth-guard support with a wireless transmitter.
Liao et al. reported a contact lens device with an electrochemical
glucose sensor, a loop antenna, a communication interface, and
readout circuitry (Fig. 5G).194 The device was powered wirelessly
using RF power and transmits information about the tear glucose
level wirelessly to an external reader. In relation to this technology,
development of a wireless swallowable capsule with
electrochemical sensors for gastrointestinal track investigation has
been a long-standing issue. Although the development of such
capsule-type devices is challenging and few experimental attempts
have been made,195 they may widely be used in clinical analyses in
the near future.
The use of smartphones has also emerged as a new trend to
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display obtained results on the spot. As an example, Mishra et
al. reported a glove-based device for defense and food security
applications (Fig. 5H).196 Electrodes were printed on a finger with
stress-enduring inks and an enzyme was immobilized for
biosensing. The device also included a swipe sampling finger and a
function for real-time wireless data transmission to a smartphone.
The number of reports related to this topic has been increasing
rapidly over the last few years. However, in most of them, the
use of the smartphone has seen major progress rather than
the improvement of sensing devices themselves. Those who
are interested in this topic should refer to some recently
published reviews.197-201
Electrochemical microfluidic components to
realize on-chip solution processing
Microfluidics is becoming an indispensable critical technique for
bio/chemical sensing. However, except for techniques such as those
involving paper-based devices, current microfluidic devices usually
rely on external bulky instruments such as pumps, valves, heaters,
etc. In particular, when the number of flow channels or reaction
chambers is large, many tubes and wires must be connected to a
chip as ironically expressed as “spaghetti syndrome.” Depending on
the applications and situations, this may be acceptable. However,
there are many other cases in which the realization of fully
integrated microfluidic devices brings benefits in improving the
utility for the users.
Many active microfluidic components, particularly micropumps
and microvalves, have been developed over the last three
decades.202-205 Among them, electrochemical microfluidic
components are suitable in promoting miniaturization and
integration, because the structure and operation of the
electrochemical components can be very simple. In addition,
because major components are electrodes that are often made of
the same materials used for sensing, there will be no or few
additional steps required for fabrication even if many sensors and
the microfluidic components are integrated. Here, we shortly
review some critical techniques in realizing electrochemical
micropumps and microvalves along with auxiliary techniques.
Electrochemical micropumps
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In many of the reported microfluidic systems, View solutions are
Article Online
transported by the application of pressure DOI:using
10.1039/C9LC01112A
an external
pressure source. To realize this on a chip, bubble-based pumps
that work by the electrolysis of water are useful. For this
purpose, a pair of electrodes such as interdigitated electrodes
are used to produce oxygen and hydrogen bubbles from both
electrodes, which then applies pressure to the solution to be
transported. Following a pioneering device by a group of
University of Twente,206 many electrolysis-based micropumps
have been reported for various purposes.26, 207-210 Because the
structure is simple, the integration of multiple pumps can be
realized easily. Nestler et al. fabricated a polymer-based device
with passive microfluidic parts and on-chip electrolytic pumps
Lab on a Chip Accepted Manuscript
(Fig. 6A).211 Although the movement of solutions is
unidirectional in these micropumps, bidirectional pumping can
also be realized employing the reduction of protons and
oxidation of hydrogen bubbles on a platinum black electrode
by applying appropriate potentials to the electrode.212 This
function will be useful to withdraw a sample solution from the
exterior. In these devices, a flexible diaphragm is sometimes
employed to separate the electrolyte solution from the
solution to be transported. Unlike cases involving other
mechanical pumps, transport of the solution is smooth with
smaller pulsations. To facilitate solution transport, the
micropump can also be coupled with a hydrophobic valve.213
Because of the easiness of integration, electrolysis-based
micropumps have been actually integrated and used to aid chemical
analyses. In these devices, reservoirs for a sample solution,
reagents, and washing buffer solutions were coupled with the
pumps and the solutions were transported to the sensing region
sequentially. Schumacher et al. developed a device with multiple
pumps and demonstrated the application to the assay of proteins
and DNAs.214 Sin et al. integrated functions including electrolysis-
based pumping, AC electrokinetic sample preparation, and
electrochemical sensing for the detection of bacterial 16S rRNA (Fig.
6B).215 Chen et al. used electrolysis-based micropumps to conduct
on-chip ELISA. Necessary reagents and washing buffer solutions
were delivered to a reaction chamber by switching the multiple
pumps sequentially (Fig. 6C).216 Geidel et al. integrated multiple
electrolysis-based pumps and a photonic sensor to demonstrate the
possibility of immunoassay. 217
Electrochemical valves
As an effective method for autonomous transport of
solutions in microfluidic devices, capillary action is
attractive.218 However, with only the capillary action,
processing solutions in a controlled manner cannot be realized.
This problem can be solved by placing switchable hydrophobic
valves at appropriate locations of flow channels. Satoh et al.
developed such valves based on direct electrowetting on a
gold electrode.219 The valve consisted of a simple electrode
pattern surrounded by a hydrophobic flow channel ceiling and
walls. A solution transported by capillary action stops there.
However, by applying an appropriate potential to the
electrode, the valve opens and the solution passes through the
valve region and moves by capillary action again. Owing to the
very simple structure, many valves could be formed and
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functioned in a network of flow channels. The valves were View Article Online
used in a device to measure the ammonia metabolism of DOI: 10.1039/C9LC01112A
hepatocytes.220 Biswas et al. reported a similar valve based on
the reductive desorption of a hydrophobic self-assembled
monolayer (SAM) of an alkanethiol from a platinum electrode
was also proposed.221 The abovementioned valves were
opened in a two-electrode configuration by the application of
a potential to the electrode that constitutes the valve. If the
three-electrode system is used for the operation and an ion-
selective electrode is used for the reference electrode, valves
that open sensing the concentration of ions can be realized.
Yamaguchi et al. reported a pH-responsive valve and a pH filter
based on this principle.222

Lab on a Chip Accepted Manuscript

EWOD has also been used to form valves for controlling capillary
action. Chen et al. used electrowetting-based valves to control the
transport of solutions by capillary action (Fig. 6D).223 The valves
were formed by forming a SAM of perfluorodecanethiol on silver
patterns. On-chip detection of Salmonella was conducted using an
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integrated three-electrode system by sandwich immunoassay.223

An electrolysis-based valve was also reported. Ezkerra et al.
proposed a bubble-based microvalve with a cantilever.224 The valve
was formed next to a flow channel and moved horizontally. When
bubbles are produced by the electrolysis of water, the valve was
closed. Leakage of gaseous pressure through a thin slit opened
the valve.

Toward automatic operation of microfluidic systems

Although the opening of the abovementioned valves was
controlled using a potentiostat, the potential of the valve
electrode can also be applied by coupling it with a redox
reaction in another control flow channel.225 Watanabe et al.
used a structure with the SAM valve mentioned above formed
in one flow channel and a zinc electrode formed in the other
flow channel (Fig. 6E).226 When the control flow channel is
empty, a solution introduced into the main flow channel stops
at the valve. The valve is opened when an electrolyte solution
introduced into the control flow channel wets the zinc
electrode. Following this, zinc is oxidized and the potential of
the electrode changes in the negative direction. This causes
the opening of the valve and the solution again stars to flow.
By forming such a pair of electrodes in appropriate locations of
flow channels, the valves can be opened in a predetermined
manner.

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pH regulation platinum auxiliary electrode that work as View an Article

Welch et al reported a device with feedback control of
solution pH.227 The device uses an extended-gate ISFET along
with a pseudo-reference electrode. The pH was regulated by
modifying flows through two input channels using integrated
pneumatic microfluidic valves. The pH control can also be
implemented electrochemically using a non-standard three-
electrode system consisting of a non-polarizable Ag/AgCl
working electrode, pH-sensitive reference electrode, and a
actuator
Online
DOI:
electrode to cause electrolysis of water.228 By10.1039/C9LC01112A
only connecting
the three-electrode system to an ordinary potentiostat, the pH
of a solution can be maintained at a predetermined constant
value by a feedback function.
Formation of a gradient for the pH or other ions is necessary
depending on the application. For this purpose, a simple
method can be the use of a pair of electrodes formed in a
small space or within a flow channel. Krabbenborg et al. used

Lab on a Chip Accepted Manuscript

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Fig. 6 Electrochemical microfluidic devices. (A) Microfluidic device with two integrated micropumps connected to a flow channel structure with a T-junction.
Adapted with permission from ref. [211]. Copyright 2010, Springer. (B) Microfluidic device with micropumps that deliver solutions to the sensing unit with a
three-electrode configuration. Adapted with permission from ref. [215]. © 2013 IEEE. (C) Microfluidic device with integrated micropumps for ELISA. (i) Layout for
the electrodes for the micropumps. (ii) Picture of the device and the magnified view of the interdigitated carbon electrodes for a micropump. Adapted with
permission from ref. [216]. Copyright 2014, American Institute of Physics under the Creative Commons Attribution 4.0 International License”. (D) Flow control
using electrowetting-based valves. (i) Integrated microfluidic chip consisting of three valves and a three-electrode system. (ii – v) Valve opening and controlled
transport of dye solutions. Adapted with permission from ref. [223]. Copyright 2015, The royal society of chemistry. (E) Device to automatically open a valve
following the injection of a solution in a control flow channel. (i) A solution to be transported stops at the valve. (ii) When the solution in the control flow channel
wets the zinc electrode, zinc is oxidized causing reductive desorption of the hydrophobic SAM of the valve. As a result, the valve opens. (iii)–(viii) Drawing of a
letter following a determined stroke order using a device with the valve and zinc electrodes formed in appropriate locations. Adapted with permission from ref.
[226]. Copyright 2017, The royal society of chemistry. (F) Droplet-based microfluidic pH regulator. Copyright permission will be obtained in the final stage.
Adapted with permission from ref. [230]. Copyright 2014, The royal society of chemistry.

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interdigitated electrodes to form µm-scale gradients of pH or
Cu+ ions employing electrolysis of water or redox reactions on
the pair of electrodes.229 Although pH at the electrodes cannot
be controlled precisely with only the pair of electrodes, the
non-standard three-electrode system mentioned above can
also be used for this purpose.228
Zhou et al. proposed a technique to form droplets with
specific pH values by microfluidics (Fig. 6F).230 The pH was
controlled by the electrolysis of water caused by the
application of current pulses. Produced hydrogen and hydroxyl
ions were separated and confined in droplets formed by the
application of pressure pulses. By adjusting the applied current
and pressure pulses, droplets of various pH values could be
formed on demand.
Strategies to improve detection sensitivity and
detection limit
Application of microelectrodes
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To improve detection performance, microelectrodes and
nanoelectrodes are often used231 because of their
advantageous features. Around the micro/nanoelectrodes,
mass transport is enhanced, which results in a high current
density. The influence of ohmic drop can also be reduced
because of the small electrode size. Furthermore, with their
small capacitances, the signal-to-background ratio and the
response time can be improved. Current generated on a single
micro/nanoelectrode is very small. Therefore, they are usually
deployed in the form of an array.
A challenging issue for microfabricated electrochemical
sensing devices is calibration. This is particularly true for
disposable devices designed for single use. In the case of
amperometric devices, the output current is influenced by the
surface state of the working electrode. Therefore, calibration is
indispensable if precision of the measurement is required.
Furthermore, without signal enhancement such as redox
cycling, amperometric detection becomes difficult as the
volume of the solution decreases, because of the rapid current
decrease caused by the depletion of analytes in the vicinity of
the working electrode. With regard to these factors,
coulometry is more beneficial. In coulometry, the output is the
charge, which is the integral of the current. Therefore, if the
sample volume is very small and the analyte is exhaustively
oxidized or reduced on the working electrode, calibration is
not necessary.232 Contrary to amperometry, the output
increases as time elapses, facilitating measurement. On the
other hand, however, the background charge increases. To
improve the sensitivity and detection limit of coulometry by
enhancing the collection efficiency of the analyte and at the
same time reducing the background charge, Sassa et al. used a
microelectrode array.233 Even if the total electrode area is the
same, the signal to the background ratio can be improved
using microelectrode arrays with thinner electrodes. To
improve the detection limit further, Ikemoto et al. converted
an analyte into silver using microelectrodes formed in separate
flow channels and measured the amount of deposited silver.234
The detection limit could be improved by more than one order
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of magnitude compared with that of the case without View Articleusing
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silver deposition. In this case, a liquid junction was used to
connect the two flow channels. The efficiency of silver
deposition could also be further improved by connecting the
analyte solution and the solution for metal deposition using a
metal junction instead of a liquid junction. By using a wire with
redox-active ends, the potentials for the redox reactions for
the analyte and silver deposition could be shifted deliberately.
For hydrogen peroxide as a model analyte, the achieved
detection limit was 2.4 nM.235
Signal amplification by redox cycling
When multiple microelectrodes are placed in close proximity,
Lab on a Chip Accepted Manuscript
chemical crosstalk by reaction products occurs. Although the
crosstalk has an adverse influence depending on the cases, it
can advantageously be used to enhance the amperometric
signal of reversible redox pairs in a technique called redox
cycling. To intentionally cause redox cycling, a pair of closely
spaced individually addressable electrodes are used. The
potential of one electrode (generator) is set at a potential that
is sufficient to oxidize (or reduce) the electroactive analyte and
the product diffuses to the other electrode (collector). The
potential of the collector is set at a potential that is sufficient
to reduce (or oxidize) the product. The products from the
collector again diffuse back to the generator, which suppresses
the depletion of reactants near the electrodes and increases
the current generated from both electrodes. Redox cycling is
also effective to minimize the influence of irreversible
electroactive interferents such as oxygen and L-ascorbic acid
because they only deplete around the generator electrode.
Since the early days of this technique, the electrode
configuration mostly used for this purpose was the
interdigitated electrodes.236 To enhance the response by
improving the collection efficiency, the distance between the
generator and collector electrodes should be as small as
possible. For this purpose, reducing the inter-electrode spacing
of the interdigitated electrodes will be a direct method.237
However, the use of electron beam lithography needs an
expensive facility and influences the productivity of the device.
To avoid this problem, Partel et al. proposed a simpler method
that involved forming nanogaps in the order of sub-100 nm
starting from structures of the micrometer scale.238 As SiO2 is
deposited, gaps between the structures are narrowed and
undercuts are formed (Fig. 7A). By only depositing gold there,
interdigitated electrodes with nanogaps are easily obtained
with the help of the undercut. With this technique, they
reported an amplification factor of 116 and a collection factor
of 99.5%.
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Lab on a Chip Accepted Manuscript

Fig. 7 Working electrode structures for redox cycling used in devices reported recently. (A) Formation and SEM image of nanogaps between interdigitated
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electrodes. Adapted with permission from ref. [238]. Copyright 2015, American Chemical Society. (B) SEM image of interdigitated electrodes. Adapted with
permission from ref. [18]. Copyright 2014, The Electrochemical Society (C) Structure and principle of operation of the electrochemical device with a nanogap.
Immobilized tyrosinase transforms inactive monophenolic molecules into electrochemically active o-quinones. These molecules subsequently undergo redox
cycling. Adapted with permission from ref. [241]. Copyright 2014 American Chemical Society. (D) Nanoring–disc electrode array. Adapted with permission from
ref. [26]. Copyright 2014, American Chemical Society. (E) Plane-recessed microdisk electrode array. Adapted with permission from ref. [247]. Copyright 2014,
American Chemical Society. (F) Inkjet-printed 3D electrode structure with a nanoporous layer between electrodes. Adapted with permission from ref. [250].
Copyright 2017, The royal society of chemistry.

Another method to improve the performance of the
interdigitated electrodes is to make them thick and three-
dimensional and use both the top surface and sidewalls of the
electrodes. Interdigitated carbon electrodes formed by the
pyrolysis of SU-8 or polyimide were fabricated for this purpose
(Fig.7B).13, 18
An effective method to alleviate this problem and achieve
appreciable performance is to use a nanogap formed between
electrodes in the direction vertical to the substrate. The
nanogap can be formed by stacking layers consisting of
electrodes, insulators, and a sacrificial layer, and dissolving the
sacrificial layer between the electrodes after the stacked
structure is completed. In this configuration, the dimensions of
the electrodes themselves can be in the order of µm.
Therefore, the structure can be made easily by
photolithography. Devices based on this structure have been
reported since the middle of 2000s.239, 240 As an application of
this technique to biosensing, Rassaei et al. used a nanogap
with electrodes and an enzyme.241 Efficient redox cycling
within the gap enabled detection of molecules of the order of
zeptomoles produced in the enzymatic reaction (Fig. 7C).
As seen in the devices that employ the nanogap, the vertically
stacked structure of electrodes and insulators facilitates the
formation of a pair of electrodes with an inter-electrode distance in
the order of nm. As a simpler structure, the electrode structure is
completed by simply introducing holes into the stacked layers and
exposing the electrodes. Ring–disk electrode arrays (Fig. 7D) 26 and
ring–ring electrode arrays 242, 243 have also been formed. Although
arrays of similar stacked electrode structures of larger dimensions
have already been reported much earlier, a recent trend has been
to form the electrode structure in an array of nanoholes. Ma et al.
used arrays of recessed ring−disk electrodes formed in a 300-nm-
deep nanochannel and detected dopamine in the presence of a
100-fold excess of ascorbic acid.244 The existence of the
nanochannel improved the selectivity of detection. They also
demonstrated the ability to differentiate reversible species with
overlapping cyclic voltammograms by adjusting the ring potential.
An interesting behavior of the recessed ring-disk nanoelectrode
array is that additional factors can contribute to signal
enhancement. The same group reported large current
enhancements in the absence of a supporting electrolyte using a
recessed ring-disk nanoelectrode array by redox cycling in
combination with ion enrichment and an unshielded ion
migration.27, 245 A current amplification as large as 2000-fold was
achieved. As a structure similar to the ring–disk and ring–ring
electrodes, Kanno et al. also used a structure with a nanogap
between electrodes formed at the periphery of a hole by the
sacrificial layer etching.246
To cause redox cycling, potentials are usually applied to the
generator and collector electrodes. However, by taking advantage
of the feature of bipolar electrodes, redox cycling can also be
generated by applying a potential to only one of the pair of
electrodes. Zhu et al. proposed the use of self-induced redox cycling
on plane-recessed microelectrodes (Fig. 7E).247 In their devices, an
array of disc electrodes were formed at the bottom of recesses and
the other electrode was formed at the top plane of an insulating
layer. When a reductive reaction occurs on the disk electrodes at
the bottom, a reverse oxidation reaction is induced at the edge of
the upper plane at the entrance of the recesses. The oxidation
current is compensated at other locations by a cathodic reaction
such as the reduction of oxygen. The recorded currents were almost
the same as the case when an oxidative potential was intentionally
applied to the top plane. Similarly, Ma et al. formed disk electrodes
at the bottom of an array of nanopores and a planar macroscopic

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top plate separated by a depth of 100 nm.248 The reaction on the
top electrode in the nanopores was coupled with the other redox
reaction at the distal end of the bipolar electrode that accompanies
changes in fluorescence or electrochemiluminescence. Thus, redox
cycling was induced between the recessed disk and the top
electrode with approximately 30× current amplification.
Although the above-mentioned devices were essentially
fabricated by the thin-film process, the necessity of an expensive
facility a facility may pose a problem. To solve this problem, beads
can be used to form a gap between the generator and collector
electrodes.249 Adly et al. used inkjet printing to form the layers of
electrodes the beads (Fig. 7F).250 Needless to say, the existence of
the beads between the electrodes hinders the mass transport of
redox species and influences the efficiency of redox cycling. Further,
the distance between the electrodes and the production yield are in
the relation of trade-off. Nevertheless, this technique will be useful
for realizing the redox cycling function in disposable devices.
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Application of nanotechnology
Detection sensitivity of these devices can be improved further by
taking full advantage of nanomaterials such as graphene, carbon
nanotubes, nanowires, and nanoparticles. A variety of techniques
have been developed for the detection of biomolecules such as
proteins and DNAs to achieve remarkable detection sensitivities.
With regard to this, numerous papers and many reviews have been
published and details are omitted here.251-257 Although the
excellence of the techniques was mainly demonstrated based on
devices of conventional sizes, they could also be incorporated in
microfabricated devices. However, a problem is that the materials
and detection chemistry involved tend to become more
complicated, making their application to microfabricated devices
more challenging.
Multiplexing of electrochemical sensors
Simultaneous multiplexed detection of analytes is required in
applications such as screening, detection of dynamic events of
neurotransmitters, and detection of proteins or DNAs.258 To realize
high-throughput multiplexed detection or real-time in situ
monitoring of high temporal and special resolutions, arraying of
sensors is a natural consequence. Detection using such arrays is
often carried out by fluorometry due to its high sensitivity and
easiness of arraying of sensors. For this technique, the realization of
large-scale sensor arrays is relatively easy as evidenced by DNA
microarrays. On the other hand, fluorometric sensor arrays also
have shortcomings such as the necessity of expensive and bulky
optical instruments, photobleaching of dyes, cross-talk between
dyes, the influence of background fluorescence, the difficulty of
measurement in turbid solutions, and the necessity of labelling for
non-fluorescent analytes. Therefore, depending on the application
and the size of the array, electrochemical sensor arrays can be a
better alternative. It is clear there are challenges in designing and
using such microarrays. The biggest problem is the operation of
multiplexed sensors and signal acquisition, particularly in large-
scale arrays. In the following section, trials to realize arrays of
sensors of relatively small numbers will be introduced.
Sensor arrays for the monitoring of cell activity
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DOI: 10.1039/C9LC01112A
neurotransmitters as target analytes. Real-time detection of
neurotransmitter release provides a direct insight into the
mechanisms of synaptic communication. Chuang et al. fabricated a
device with microelectrodes and interdigitated generator/collector
electrodes for the simultaneous detection of action potential and
dopamine release from PC12 cells.259 Picollo et al. used an array of 4
× 4 electrodes for amperometric and voltammetric detection of
exocytosis from chromaffin cells and adrenal slices.260 Wang et al.
formed an array of platinum microelectrodes in a PDMS chamber to
detect dopamine released from PC12 cells (Fig. 8A).261 The cells
were cultured in the PDMS chamber coated with collagen. The
same group fabricated arrays of individually addressable
Lab on a Chip Accepted Manuscript
microelectrodes coated with collagen formed in an SU-8 microwell
for the detection of neurotransmitter release from a single PC12
cell.262 Imaging of the spatial distribution of exocytosis was also
demonstrated. Yakushenko et al. fabricated a device for parallel
real-time recording of vesicular neurotransmitter release using 64
microelectrodes with a minimum electrode diameter of 3 μm (Fig.
8B).263
Kisler et al. fabricated transparent microelectrode arrays for
simultaneous amperometric measurements and fluorescence
imaging (Fig. 8C).264 Fluorescence images of chromaffin cells were
taken by total internal reflection fluorescence microscopy while the
release of catecholamine from single vesicles was measured by
amperometry. Carabellia et al. demonstrated the simultaneous
detection of exocytotic events of chromaffin cells upon stimulation
with KCl using an array of microelectrodes of boron-doped
nanocrystalline diamond.265 Wigström et al. fabricated a
microelectrode array consisting of 16 1.2-μm-wide platinum band
electrodes (Fig. 8D).266 Catecholamine released from bovine
chromaffin cells by single exocytotic events was detected
amperometrically. As for the detection of exocytosis from cells
using microelectrode arrays (MEAs), a review was published in
2015.267 Readers should refer to this for more information about
devices other than those introduced here.
As a practically important application, Besant et al. used sensor
arrays for the measurement of antibiotic susceptibility of bacteria
(Fig. 8E).268 They used miniaturized wells to capture, concentrate,
and analyze bacteria. The metabolic activity of the bacteria was
measured electrochemically using a redox-active molecule.
Small arrays of sensors for proteins and DNAs
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Fig. 8 Integrated electrochemical devices for analysis of cell activity. (A) Array of 36 addressable microelectrodes for spatial measurements of single vesicle
release. Adapted with permission from ref. [261]. Copyright 2013, American Chemical Society. (B) Device for parallel detection of neurotransmitter release. (i)
Array of 64 microelectrodes. (ii) Simultaneous amperometric recordings of spontaneous exocytotic events obtained using 10 microelectrodes. Adapted with
permission from ref. [263]. Copyright 2013, American Chemical Society. (C) Transparent ITO electrode for simultaneous amperometric detection of exocytosis and
fluorescence microscopy. (i) ITO (top) and gold (bottom) electrodes with four (left) and single (right) electrode geometries. (ii) Correlation of results obtained by
amperometry and total internal reflection fluorescence imaging for a vesicle release event from a chromaffin cell. Adapted with permission from ref. [264].
Copyright 2012, Scientific Research Publishing Inc. under the Creative Commons Attribution 4.0 International License. (D) Device with 16 microelectrodes for
detection of exocytosis detection. (i) Side view of sensing region, where the probe is in touch with a cell. (ii-iii) SEM and microscopy images of the microelectrode
array. Adapted with permission from ref. [266]. Copyright 2016, American Chemical Society. (E) Array of microwells with electrodes for phenotypic profiling of
antibiotic-resistant bacteria. Adapted with permission from ref. [268]. Copyright 2011, The royal society of chemistry. (F) Impedimetric immunosensor array for
the detection of tumor marker proteins. Adapted with permission from ref. [269]. Copyright 2011, The royal society of chemistry. (G) Array of 96 sensing sites
formed on a printed circuit board for screening of an enzyme. Adapted with permission from ref. [270]. Copyright 2013, American Chemical Society. (H) Device
for digital protein detection with an array of wells for stripping analysis of enzyme-induced metallization. Adapted with permission from ref. [271]. Copyright
2016, American Chemical Society.
For cases such as diagnosis of cancer, speciation of infectious reported a device with nine chambers each containing a circular
diseases, and profiling of antibiotic-resistance, detection of multiple gold working electrode surrounded by a platinum auxiliary
marker proteins is required. Arrays of electrochemical sensors can electrode (Fig. 8F).269 Ag/AgCl reference electrodes were inserted
be constructed for this purpose. Unlike DNA chips, a challenge is through a sealing lid. A tumor marker, tenascin C, was detected
the preparation of probe molecules. Therefore, the number of impedimetrically following the formation of protein-antibody
sensors integrated in such devices is limited for the detection of a complexes, peroxidase-catalyzed production of the precipitate, and
single or a small number of target proteins, although the possibility redox reactions of ferri-/ferrocyanide. Abdellaoui et al. fabricated a
of the integration of many sensors was demonstrated. Steude et al. device with 96 wells (Fig. 8G).270 A pair of a carbon working

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electrode and a Ag/AgCl electrode as the auxiliary/reference
electrodes were formed in each well by screen-printing. Screening
of laccase from the fungus Trametes versicolor was conducted. The
device was also used to identify Laccase inhibitors for the enzyme,
pH optima, and interfering molecules.270 Wu et al. formed a
microelectrode array to detect alkaline phosphatase (ALP) secreted
from liver cancer cells by digital analysis (Fig. 8H).271 The product of
the enzymatic reaction of ALP reduced Ag+ to silver. Stripping
analysis was conducted for the deposited silver.271 Luo et al. used a
sensor array with cross-linked poly(ethylene glycol) for
impedimetric detection of insulin and CRP.272 PEG reduced
unwanted interaction with proteins effectively.
Kling et al. used a unique mechanism to detect up to eight
antibiotics.273 The detection was based on a natural sensing system
including an operator DNA and a repressor protein, which evolved
in certain bacteria to resist antibiotics. The operator DNAs were
immobilized on the surface of a flow channel and the repressor
proteins were bound to the DNA. In the absence of antibiotics, the
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repressor proteins are tightly bound to the operator DNA. However,
when antibiotics are present, the repressor proteins detach from
the operator. Changes in the reaction of enzymes conjugated to the
repressor proteins were detected by amperometry.
In realizing multiplexed detection of proteins in real samples,
detection limits must also be sufficiently low. In many cases,
additional amplification is needed for this purpose. Rusling et
al. proposed methods to realize very low detection limits. In
one method, they used detection antibodies with polymerized
horseradish peroxidase (HRP) labels for the detection of
interleukin-6 (IL-6).274 Otherwise, they used magnetic beads
massively loaded with enzyme labels and secondary antibodies
for protein sensor arrays and have reported very low detection
limits.275-277 They further used this technique in a device with
an array of 32 microelectrodes, which was further multiplexed
with an 8-port manifold to allow 256 measurements.278
As for large arrays of DNA sensors, the CMOS technology
mentioned later will be indispensable. Depending on the
application, small arrays are also used. Pursey et al. fabricated a
multi-sensor chip that detects three specific bladder cancer DNA
markers simultaneously.279 Their DNA probe had a hairpin structure
with a porphyrin marker at the end, which became
electrochemically active accompanying hybridization with the
target strand through a structural change. Slinker et.al. fabricated
an array of 16 DNA-modified electrodes for multiplexed
detection of DNAs and DNA binding proteins.280 Detection using
the device relied on DNA mediated charge transfer.
Decreasing number of electrical connections in
multiplexed sensing devices
In applications such as the high-throughput speciation of
infectious diseases, antibiotic-resistance profiling, and
screening of cell libraries, large arrays of individually
addressable sensing components are required. Compared with
optical methods, this is really a challenge with regard to
operation and data acquisition, because electrodes may need
to be connected to external instruments through a large
number of contact pads. For devices with the number of
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working electrodes in the order of tens or hundreds,
View Articlesome
Online
effective techniques have been proposed.DOI: 10.1039/C9LC01112A
Use of matrix configuration for electrodes and flow channels
To mitigate the above-mentioned difficulty, layouts of electrodes,
flow channels, and reaction chambers in the form of a matrix have
been proposed. A group of Tohoku University proposed an
electrode layout consisting of an n × n array of a pair of working
electrodes for redox cycling (Fig. 9A). Electrodes in each row and
column are connected. When a row and a column are selected and
potentials are applied to the electrodes, only the sensing site
corresponding to the selected row and column is activated.
Therefore, only 2n bonding pads are sufficient for connection to an
Lab on a Chip Accepted Manuscript
external instrument to activate n2 sensing sites. The group
demonstrated operation of up to 1024 (32 × 32) addressable
sensing sites.281 They used this technique in devices for evaluation
of embryoid bodies through their alkaline phosphatase activity,246,
282 detection of alkaline phosphatase secreted by HeLa cells,283 DNA
assay.30 Kätelhön et al. also used two sets of 16 parallel band
electrodes formed perpendicularly to each other to cause redox
cycling and detect current in a nanocavity with a height of 65 nm.284
Lam et al. used bands of reference and auxiliary electrodes
formed along flow channels and bands of working electrodes
formed orthogonally to the flow channels.285 In the presence of
solutions, the electrical circuit was complete with selected working,
reference, and auxiliary electrodes. The device was used for the
detection of pathogenic bacteria and antibiotic-resistance markers.
Zhang et al. further used a technique to separate solutions. They
formed a set of gold band pseudo-reference and auxiliary
electrodes and another set of gold band working electrodes formed
orthogonally to make 400 (20 × 20) sensing sites (Fig. 9B).286 Each
site covered by the three-electrode system was covered with a
porous wettable square each separated by hydrophobic barriers.
Application of bipolar electrochemistry
Bipolar electrochemistry provides another solution for a
high number of sensors being required.287-291 In orthodox
devices, small rectangular electrodes (bipolar electrodes: BPE)
are placed in a solution between two driving electrodes (open
BPE). In bipolar devices, the electrodes are operated wirelessly
and simultaneously without leads for connection to external
instruments. (Fig. 9C) shows the principle of detection for the
open bipolar system. When a voltage is applied to the driving
electrodes, a potential drop is generated in the solution
between the driving electrodes. For a sufficiently low ionic
strength, a uniform potential gradient is expected. As a result,
an interfacial potential difference develops between the
electrode and the solution, and the potential difference
depends on the location on the electrode and is the largest at
both poles. If the potential difference is sufficiently large at the
cathodic and anodic poles, reduction and oxidation occur,
respectively, in the presence of redox compounds. Because the
BPEs are electrically isolated from external instruments,
outputs cannot be obtained in the form of a current. Therefore,
optical methods are alternatively used to obtain the
information on redox reactions, and
electrochemiluminescence (ECL), particularly based on the
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Ru(bpy)32+/tripropylamine (TPA) system, is often used. An
advantage of ECL is that it does not need external light
sources. Therefore, the background can be minimized and
detection can be highly sensitive. Although the Ru(bpy)32+/TPA
system is predominantly used in bipolar devices, coupling the
anodic or cathodic reactions to an enzymatic reaction needs
consideration. For this purpose, the luminol/H2O2 ECL system
may alternatively be used to couple the reaction with
enzymatic reactions that produce H2O2.292 A great advantage
of the open bipolar system is that the structure of each BPE
can be very simple and a large array of BPEs can easily be
realized. Chow et al. demonstrated the simultaneous
operation of 1000 BPEs. 293 (Fig. 9D)
principle of detection is essentially different. In closed bipolar
View Article Online
systems, the sum of all the interfacial DOI: 10.1039/C9LC01112A
potential differences
between the electrodes and the solutions and ohmic drop in
the solutions is equal to the driving voltage. Therefore, closed
bipolar devices can be operated with a relatively small applied
voltage of approximately 1 V.
The closed BPEs can effectively be used for imaging.
Guerrette et al. used a large array of BPE and separated the
analyte solution and the other resazurin solution for imaging
(Fig. 9E).296 The analyte on the anode side was detected by
fluorescence emitted from the reaction product on the
cathodic pole. Oja et al. demonstrated two- and three-
dimensional imaging of concentration profiles in diffusion

Lab on a Chip Accepted Manuscript

In open bipolar ECL devices, the background signal from the
driving electrode poses a problem. In addition, the reactants
for the anodic and cathodic reactions must be separated
depending on the situation. To solve these problems,
compartments for the anodic and cathodic reactions have
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layers of carbon fiber microelectrodes over an array of BPEs.297
3-dimensional imaging was realized by collecting 2-
dimensional images at different axial positions. Although these
devices are not necessarily fabricated by photolithography or
other related microfabrication techniques, they demonstrate a

been separated and solutions have been connected with a
BPE.294, 295 The configuration in which BPEs are in contact with
solutions of different compartments is called a closed bipolar
system. The open and closed bipolar systems may be
considered as simple structural variations. However, their
promising application of this technique. This technique was
also used for multiplexed detection. Wu et al. formed an array
of reactions and sensing channels connected by BPEs for the
multiplex detection of cancer biomarkers (Fig. 9F).298 Aptamers
or antibodies and an electroactive tag synthesized by doping a

Fig. 9 Techniques to reduce number of electrical connections. (A) Crossbar electrode array in the form of a matrix. Adapted with permission from ref. [281].
Copyright 2011, The royal society of chemistry. (B) Array of individually addressable electrochemical cells with crossbar electrodes. Adapted with permission from
ref. [286]. Copyright 2017, American Chemical Society. (C) Principle of detection of the open bipolar system. (i) Reactions that occur at the anodic and cathodic
poles. (ii) Variation of the potential in the solution and polarizations at the anodic and cathodic poles. Adapted with permission from ref. [289]. Copyright 2013,
Wiley-VCH Verlag GmbH & Co. KGaA. (D) Open bipolar device with a large array of BPEs (i) and ECL observed from the BPEs (ii). Adapted with permission from ref.
[293]. Copyright 2009, American Chemical Society. (E) 2D array of closed bipolar electrodes for fluorescence imaging. Adapted with permission from ref. [296].
Copyright 2013, American Chemical Society. (F) Array of closed bipolar electrodes for the detection of cancer biomarkers. Adapted with permission from ref.
[298]. Copyright 2015, American Chemical Society. (G) Light-addressable potentiometric sensor with a light source coupled with an X-Y stage. Reprinted with
permission from ref. [311]. Copyright 2012, Elsevier.

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large number of thionine molecules in silica nanoparticles
were used for the cathodes.298 Wang et al. reported a
multicolor ECL device based on closed BPE employing the
potential dependence of the emission color of
electrochemiluminophores.299 Multicolor (green−yellow−red)
ECL was realized using a mixture of [Ru(bpy)3]2+ and [Ir(ppy)3]
with TPA as the co-reactant. The device was used for the
quantification of prostate-specific antigen (PSA). A PSA
concentration-dependent silver bridge was made in the gap of
the BPEs to modulate the conductivity. The color change
depended on the concentration of PSA.
Open bipolar systems are promising to realize large-scale sensor
arrays for analytes such as DNAs.300 As a trend, reported devices
have been shifting to closed bipolar systems. This may be
reasonable considering that they can solve the problems of the
open bipolar systems. On the other hand, because of the additional
structure that separates solutions, the operation of a large array of
BPE that can easily be achieved using the open bipolar systems 293
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becomes difficult. A drawback of the BPE devices is that they must
rely on reporting techniques such as ECL. In this respect, for a
limited number of integrated BPEs, the superiority over the
conventional three-electrode systems that measure current directly
is not convincing.
Nevertheless, the technique used in closed bipolar systems
provides a solution for a difficult problem in miniaturizing
electrochemical systems, which often require liquid junctions.
However, microfabricating good liquid junctions without an ohmic
drop is really challenging. With regard to this, closed bipolar
systems demonstrate that solutions can also be connected using
metal junctions.301, 302 The technique will facilitate designing more
sophisticated electrochemical microsystems.
Light-addressable electrochemical devices
The use of light also solves the problem of selective signal
acquisition. Light-addressable electrochemical devices are
operated based on this method. One of the critical advantages
of the devices is that only a thin sensing plate is sufficient and
no fine patterns as seen in microelectrode arrays are
necessary. The light-addressable potentiometric sensor (LAPS)
is one such device.303-305 The sensing part of the LAPS consists
of an electrolyte–insulator–semiconductor structure. A change
in the potential difference at the electrolyte–insulator
interface changes the thickness of the depletion layer in the
semiconductor. When a location is illuminated in this state,
electron–hole pairs are generated and separated in the
depletion layer, which then generates photocurrents. The
illuminated locations become independent sensing sites.
Therefore, an array of sensors can be realized with only a
simple structure. The LAPS can also be used for chemical
imaging. As mentioned later, such image sensors can also be
realized by the CMOS technology in the form of a large array of
ISFETs integrated with other electronic components. However,
the number and locations of pixels are unchangeable, which
may cause inconvenience in cases where the location of the
molecules or objects is unpredictable. The problem can be
alleviated in the case of the LAPS. On the other hand, however,
a large sensing area and the length of time for measurement
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can be trade-offs. Furthermore, a light source,Viewa Article focusing
Online
DOI: 10.1039/C9LC01112A
optics, and a scanning mechanism are additionally required.
To solve the problem related to signal transduction, some
alternative techniques have been used. One such technique
used a fixed array of light-emitting diodes (LEDs) or lasers to
illuminate multiple regions on the sensing surface.306, 307 The
photocurrent generated at each spot is modulated with the
frequency of the corresponding LED. The total measured
photocurrent is a superposition of the local photocurrents. The
photocurrent derived from a single spot can be obtained by a
fast Fourier transform.306 An organic light-emitting diode panel
was also used for high-resolution imaging. Miyamoto et al.
used this device to demonstrate pH imaging with 135 frames
Lab on a Chip Accepted Manuscript
per second (fps).308 Alternatively, a bundle of optical fibers was
used to achieve guided light spots flexibly. Miyamoto et al.
achieved two-dimensional pH imaging in a flow channel at a
maximum rate of 100 fps.309 The combination of an optical
chopper and microlens array can also realize multi-spot
interrogation using a single beam.310 The digital micromirror
device (DMD) used in video projectors is also effective in
increasing the number of light spots. It consists of a large array
of micromirrors that can be flipped individually, which enables
the rapid production of a large number of light patterns.
Wagner et al. employed this technique to readout (Fig. 9G).311
The LAPS is effective for monitoring the metabolic activities of
living cells. Wagner et al. recorded the metabolic activity of
macrophages at 16 measurement spots for the concentration
gradients of a drug.312 Shaibani et al. conducted a real-time
measurement of cancer cell metabolism and their response to
anticancer drugs by detecting local changes in the pH as cancer cells
released lactate.313 The same group used pH-sensitive hydrogel
nanofibers for the sensitive coating of LAPS to detect pH changes
caused as a result of sugar fermentation by E. coli.314 Su et al.
fabricated a device with the LAPS and an impedance sensor for
simultaneous monitoring of extracellular acidification and growth of
HeLa cells.9 Hu et al. used LAPS coupled with a microfluidic system
to monitor cellular metabolism and drug effect.315 Human breast
cancer cells MCF-7 were used and real-time monitoring of
extracellular acidification was conducted rapidly and sensitively
under physiological conditions. Although the light beam was not
necessarily scanned and only the pH-sensing function was used in
these cases, 2D detection was also conducted. Takenaga et al.
observed the metabolic activity of Chinese hamster ovary and
tumor cells.307 They used a 4×4 array of infrared LEDs as the light
source. The pH changes accompanying the addition of glucose and
changes in metabolic activity were measured. Wu et al. used LAPS
for the 4×4-spot label-free detection of DNA using a 4×4 array of
infrared LEDs.306 Although the LAPS was developed to detect the pH
in the initial stage, it can be sensitive to other ions and molecules
such as DNA.304 Du et al. used bioengineered olfactory sensory
neurons to make the LAPS respond to odorant molecules.308
The LAPS has an electrolyte–insulator–semiconductor structure
and detects changes in the potential difference at the electrolyte–
insulator interface. In other words, detection of the progress of
redox reactions is not possible due to the presence of an insulator
layer. However, by removing it and using an appropriate
semiconductor surface, amperometric detection is also possible.316
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Choudhury et al. used this technique and demonstrated multi-spot
detection of redox reactions on a planar hydrogen-terminated
silicon electrode modified with well-defined organic monolayers.317
An n-type semiconductor can be used for oxidation, whereas a p-
type semiconductor can be used for reduction. They also
demonstrated the applicability of their device to the detection of
DNA hybridization. Seo et al. used hematite (α-Fe2O3) for the anode
for the selective detection of oxidizable biomolecules.318 They
demonstrated quantitative analysis of dopamine.
Higher-level integration by CMOS technology
In the previous section, techniques for multiplexed
detection were introduced. However, as the number of
The ISFETs directly detected pH changes caused by the release
View Article of
Online
protons during nucleic acid polymerization.DOI:
The10.1039/C9LC01112A
amplification and
measurement started with the binding of the complementary
primer to its target sequence, and no changes were observed for a
mismatch primer.
Devices directed to chemical imaging have also been
developed. Hayasaka et al. fabricated an array of 20 × 20
boron-doped diamond microelectrodes for the real-time
imaging of histamine (Fig 10E).327 The electrodes were formed
on a separate substrate and transferred to another CMOS LSI
wafer. Rothe et al. integrated an array of 32 × 32 platinum
black working electrodes, on-chip reference and auxiliary
electrodes, and potentiostats (Fig. 10F).328 The device could

Lab on a Chip Accepted Manuscript

sensors increases to the order of hundreds, thousands, or
detect faradaic currents that were more than 10,000 times
more, the application of these techniques will be difficult. This
smaller than charging currents without a Faraday cage.328
naturally leads to the integration of necessary components
Electrochemical images could be acquired at up to 90 fps.
including sensors and circuits for operation and signal
Bellin et.al. fabricated a device with an array of 1,824
processing by the CMOS processes. By amplifying and digitizing
electrodes multiplexed to 38 parallel output channels for the
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signals on the same chip, noise interference originating from

imaging of phenazine metabolites produced from biofilms of
lengthy signal paths can also be avoided, enabling
Pseudomonas aeruginosa.329 They demonstrated potential-
measurements without additional shielding such as that by a
sweep-based electrochemical imaging. Nemeth et al.
Faraday cage. Although CMOS devices are the most
fabricated a high-speed ion-imaging device with 64 x 64 pixels
sophisticated among devices treated in this review, trials to
consisting of an array of ISFETs.330 The device can display the
integrate components other than sensors can be found even in
distribution of protons at approximately 100 fps. The same
the 1980s and some reviews have been published.74, 319, 320
group also reported another imaging device with 16 x 16 pixels
During the last decade, the level of integration of CMOS-based
that operates at 333 fps.331 Sawada et al. reported various
electrochemical sensing devices advanced further and very
charge-coupled device (CCD)-type chemical imaging devices
sophisticated devices that may be called systems-on-a-chip
with an array of pH-ISFETs including one with 128 x 128
have been reported.
pixels.332 (Fig. 10G). They also reported applications in imaging
As examples of multiplexed detection, Kim et al. fabricated a
of the distribution of the concentration of potassium ions or
sensor array consisting of 100 platinum electrodes for the
acetylcholine.
simultaneous amperometric detection of neurotransmitter release
Unlike physical sensors, chemical sensors often contain
from chromaffin cells (Fig. 10A).321 The electrode array was
unstable components such as biomolecules and the Ag/AgCl
operated using integrated potentiostats with time-division
reference electrode. Among them, a troublesome component
multiplexing to limit the number of outputs. Inoue et al. fabricated
to be considered is the reference electrode. As mentioned
an array of 400 measurement points for electrochemical imaging
previously, Ag/AgCl electrodes formed by the thin-film process
and multi-point biosensing (Fig. 10B).322 The device had a light-
are very easily damaged. Therefore, it would imprudent to try
shield structure to reduce noise generated by a photocurrent and a
and integrate them onto the chip. For amperometric devices, a
mode-selection function. Jafari et al. developed a 54-channel device
realistic solution will be to use a pseudo-reference electrode
that conducts fast-scan cyclic voltammetry and calibration for DNA
fabricated from noble metals. For potentiometric devices, this
analysis.323 The device contained 600 time-multiplexed DNA sensors
does not work. In this case, the reference electrode should be
and 54 pH sensors along with an arbitrary waveform generator, a
separated as an independent module. In this case, the size of
memory, a current conveyer, an analog-to-digital converter (ADC),
the module can be comparable with the size of the sensing
and a wireless transmitter. The function of the device was validated
chip and the use of screen-printed thick-film electrodes will
by detecting prostate cancer DNA and the achieved detection limit
alternatively be used.
was 10 aM. Ganguli et al. fabricated an array of 1024 × 1024 ISFETs
with immobilized peptide nucleic acid (PNA) probes for label-free
sensing of nucleic acids (Fig. 10C).324 They detected miRNA Let-7b of
concentrations as low as 1 nM. Rothberg et al. fabricated an array
of 1.2 million ISFET pH sensors for DNA sequencing (Fig. 10D).325
The sequence was determined by sensing ions produced by
template-directed DNA polymerase synthesis. Sequencing of
bacterial genomes was demonstrated using this device. Toumazou
et al. demonstrated label-free real-time amplification and detection
by polymerase chain reaction (PCR) and loop-mediated isothermal
amplification using a device containing 40 ISFETs, 10 temperature
sensors, resistive heaters, signal processing and control circuitry.326

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Lab on a Chip Accepted Manuscript

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Fig. 10 Highly integrated electrochemical sensing devices fabricated by CMOS process. (A) 10 × 10 array of working electrodes with in-pixel CMOS potentiostats.
for detection of exocytosis of chromaffin cells. (i) Photograph of the chip. (ii) Magnified views of an element obtained at different focal planes. Reprinted with
permission from ref. [321]. Copyright 2013, Elsevier. (B) Amperometric bioimaging chip. (i) Array of 400 individually addressable electrodes for imaging of
distribution of O2 and H2O2. (ii) Color maps of distribution of current before and after injection of H2O2 and Na2SO3 solution. Adapted with permission from ref.
[322]. Copyright 2015, The royal society of chemistry. (C) 1024 x 1024 ISFET array for label-free microRNA detection. Adapted with permission from ref. [324].
Copyright 2018, Springer. (D) DNA sequencing chip with 1.2 million wells. (i) Structure of a well and principle of sequencing. (ii) An 8” wafer with ~200 chips. (iii)
Picture of a chip. Adapted with permission from ref. [325]. Copyright 2011, Springer Nature Limited (E) Array of boron-doped diamond microelectrodes for
amperometric sensing. Adapted with permission from ref. [327]. ©2009 IEEE (F) High-speed chemical imaging sensor array with 1024 Pt or Pt black working
electrodes. (i) Platinum electrodes with different diameters. (ii–iv) SEM images of Pt and Pt black electrodes. Adapted with permission from ref. [328]. Copyright
2014, American Chemical Society. (G) Images of pH recorded by the device and corresponding optical image when a buffer solution was additionally dropped
onto the sensitive area of the device. (i) Initial state (pH 9.01). (ii) After a solution of pH 4.01 was dropped. (iii) Diffusion of the solution. Adapted with permission
from ref. [332]. Copyright 2014, The IEICE (H) Implantable wireless glucose sensor chip. Reprinted with permission from ref. [333]. Copyright 2019, Elsevier. (I)
Wireless sensing chip for alcohol monitoring injectable using a syringe. Adapted with permission from ref. [334]. ©2009 IEEE (J) CMOS chip for electrochemical
detection embedded in a microfluidic device. Adapted with permission from ref. [335]. Copyright 2013, The royal society of chemistry.

An issue with the highly integrated CMOS devices will be the

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fabrication cost. When a very large number of sensors need to
be used, the integration of sensors and other circuits is a
natural consequence. On the other hand, in applications where
the cost for analysis must be low, they will not be competitive
enough compared with the opposite extreme such as
disposable paper-based devices. While investigating previous
CMOS-based devices, we found devices with only a few
sensors integrated with operational circuits such as those for
potentiostats. Although these may suggest a step toward more
sophisticated devices, we wonder if they can be replaced with
simpler devices such as the paper-based devices operated
using external instruments. Even in such cases, cost reduction
could be achieved by decreasing the chip size and increasing
will include devices for environmental monitoring,View implantable
Article Online
devices, and devices directed to basicDOI: 10.1039/C9LC01112A
research that use
animals. Furthermore, the use of smartphones as a platform
for the collection of data is increasing,197, 336-339 which suggests
that applicable fields for electrochemical sensing devices will
expand further.
Acknowledgments
We thank Dr. Yoshitaka Yoshizumi and Dr. Isa Anshori for their
support in completing this review. This work was partially
supported by JSPS KAKENHI Grant Number 19H05680.

Lab on a Chip Accepted Manuscript

the number of chips obtained from a wafer. Mujeeb et al.
developed a CMOS-based implantable chip with a size of only References
1.4 x 1.4 mm2 offering functions such as amperometric sensing, 1. D. Huckle, Expert Rev. Mol. Diagn., 2008, 8, 679 - 688.
signal processing, and power transfer. In vivo tests were 2. P. B. Luppa, C. Müller, A. Schlichtiger and H. Schlebusch,
conducted employing the device on nondiabetic Sprague Trends Anal. Chem., 2011, 30, 887 - 898.
Published on 13 February 2020. Downloaded on 2/15/2020 1:20:42 AM.

Dawley rats and their blood glucose level was monitored (Fig. 3.
10H).333 Jiang et al. integrated a pH sensor, an alcohol sensor, a
potentiostat, a signal processing circuit, a signal transmitter, and a 4.
power receiving coil on a chip of 0.85 × 1.5 mm2 (Fig. 10I).334
In coupling a CMOS chip to microfluidic structures, the difference 5.
in the fabrication process and chip size poses a problem. To cope
6.
with this problem, Huang et al. embedded a chip of 1.8 mm × 1.6
mm in a micromachined silicon carrier. By leveling the surfaces of
the chip and the carrier to within 100 nm, an obstacle-free flat 7.
smooth surface was realized (Fig. 10J).335 The flat surface facilitated
the formation of thin-film metal interconnects between the contact
pads of the chip and the perimeter of the carrier and coupling of
SU-8 microfluidic mixers and detection channels. 8.
Conclusions and outlook 9.
Compared with bio/chemical sensing devices based on other
detection principles, electrochemical devices offer advantages 10.
in terms of miniaturization, batch fabrication, and integration
of the same and/or different components. A wealth of
techniques developed toward electrochemical sensing, along 11.
with nanotechnology, can be implemented to improve the 12.
detection sensitivity and lower the detection limit. On the
other hand, by merging the techniques with those of 13.
microfluidics, additional functionalities can be incorporated for
the processing of solutions. 14.
Required functions for these devices are becoming more
diverse. For POCT, devices must be very inexpensive, 15.
disposable, and user-friendly. In addition, such devices must
provide precise reliable data without any errors. Wearable 16.
devices must meet requirements such as wireless operation
17.
and data transmission, resistance to deformation, and
biocompatibility. Highly integrated CMOS devices with very 18.
large-scale electrode arrays will be useful for the collection of
big data in bioscience.
To minimize the inconvenience of users, wireless data 19.
transmission is becoming mandatory for some applications.162
Although the technique is mostly applied in wearable devices, 20.
applications that need this option will increase. Some of these
N. P. Pai, C. Vadnais, C. Denkinger, N. Engel and M. Pai,
PLoS Med., 2012, 9, e1001306.
W. Jung, J. Han, J.-W. Choi and C. H. Ahn, Microelectron.
Eng., 2015, 132, 46 – 57.
S. K. Vashist, P. B. Luppa, L. Y. Yeo, A. Ozcan and J. H. T.
Luong, Trends Biotechnol., 2015, 33, 692 - 705.
N. Yang, S. Yu, J. V. Macpherson, Y. Einaga, H. Zhao, G.
Zhao, G. M. Swain and X. Jiang, Chem. Soc. Rev., 2019, 48,
157 - 204.
A. Sbartai, P. Namour, A. Errachid, J. Krejči, R. Šejnohová,
L. Renaud, M. L. Hamlaoui, A.-S. Loir, F. Garrelie, C.
Donnet, H. Soder, E. Audouard, J. Granier and N. Jaffrezic-
Renault, Anal. Chem., 2012, 84, 4805 - 4811.
W. Smirnov, N. Yang, R. Hoffmann, J. Hees, H. Obloh, W.
Müller-Sebert and C. E. Nebel, Anal. Chem., 2011, 83,
7438 - 7443.
R. Oyobiki, T. Kato, M. Katayama, A. Sugitani, T.
Watanabe, Y. Einaga, Y. Matsumoto, K. Horisawa and N.
Doi, Anal. Chem., 2014, 86, 9570 - 9575.
M. B. Joseph, E. Bitziou, T. L. Read, L. Meng, N. L. Palmer,
T. P. Mollart, M. E. Newton and J. V. Macpherson, Anal.
Chem., 2014, 86, 5238 - 5244.
R. L. McCreery, Chem. Rev., 2008, 108, 2646 - 2687.
F. Liu, G. Kolesov and B. A. Parkinson, Anal. Chem.,
2014, 86, 7391 - 7398.
R. R. Kamath and M. J. Madou, Anal. Chem., 2014, 86,
2963 - 2971.
Y. Lim, J.-I. Heo and H. Shin, Sens. Actuators B, 2014, 192,
796 - 803.
R. A. Saylor, E. A. Reid and S. M. Lunte, Electrophoresis,
2015, 36, 1912 - 1919.
A. K. Dengler, R. M. Wightman and G. S. McCarty, Anal.
Chem., 2015, 87, 10556 - 10564.
J. Lee, D. Sharma, Y. Lim and H. Shin, Sens. Actuators B,
2018, 267, 467 - 475.
M. Morita, K. Hayashi, T. Horiuchi, S. Shibano, K.
Yamamoto and K. J. Aoki, J. Electrochem. Soc., 2014, 161,
H178 - H182.
A. Ambrosi and M. Pumera, Chem. Soc. Rev., 2016, 45,
2740 - 2755.
L. Soleymani, Z. Fang, X. Sun, H. Yang, B. J. Taft, E. H.

This journal is © The Royal Society of Chemistry 20xx J. Name., 2013, 00, 1-3 | 25

Please do not adjust margins

 Lab
Please do onadjust
not a Chipmargins Page 26 of 32

ARTICLE Journal Name

Sargent and S. O. Kelley, Angew. Chem. Int. Ed., 2009, 48,
8457 - 8460.
21. J. Das and S. O. Kelley, Anal. Chem., 2011, 83, 1167 - 1172.
22. E. Vasilyeva, B. Lam, Z. Fang, M. D. Minden, E. H. Sargent
and S. O. Kelley, Angew. Chem. Int. Ed., 2011, 50, 4137 -
4141.
23. P. Podešva, I. Gablech and P. Neužil, Anal. Chem., 2018,
90, 1161 - 1167.
24. H. Li and N. Wu, Nanotechnology, 2008, 19, 275301.
25. J. Hees, R. Hoffmann, A. Kriele, W. Smirnov, H. Obloh, K.
Glorer, B. Raynor, R. Driad, N. Yang, O. A. Williams and C.
E. Nebel, ACS Nano, 2011, 5, 3339 - 3346.
26. C. Ma, N. M. Contento, L. R. G. II and P. W. Bohn, ACS
Nano, 2013, 7, 5483 - 5490.
49. R. Seemann, M. Brinkmann, T. Pfohl and S. View Herminghaus,
Article Online
Rep. Prog. Phys., 2012, 75, 016601.DOI: 10.1039/C9LC01112A
50. T. Schneider, J. Kreutz and D. T. Chiu, Anal. Chem., 2013,
85, 3476 - 3482.
51. D. Itoh, E. Koyachi, M. Yokokawa, Y. Murata, M. Murata
and H. Suzuki, Anal. Chem., 2013, 85, 10962 - 10968.
52. V. Linder, S. K. Sia and G. M. Whitesides, Anal. Chem.,
2005, 77, 64 - 71.
53. F. Yang, X. Zuo, Z. Li, W. Deng, J. Shi, G. Zhang, Q. Huang,
S. Song and C. Fan, Adv. Mater., 2014, 26, 4671 - 4676.
54. O. Ordeig, P. Ortiz, X. Muñoz-Berbel, S. Demming, S.
Büttgenbach, C. Fernández-Sánchez and A. Llobera, Anal.
Chem., 2012, 84, 3546 - 3553.
55. Y. Li, C. Sella, F. Lemaître, M. Guille-Collignon, C. Amatore

Lab on a Chip Accepted Manuscript

27. C. Ma, W. Xu, W. R. A. Wichert and P. W. Bohn, ACS Nano, and L. Thouin, Anal. Chem., 2018, 90, 9386 - 9394.
2016, 10, 3658 - 3664. 56. I. A. Ges, R. L. Brindley, K. P. M. Currie and F. J.
28. M. Hüske, A. Offenhäusser and B. Wolfrum, Phys. Chem. Baudenbacher, Lab Chip, 2013, 13, 4663 - 4673.
Chem. Phys., 2014, 16, 11609 - 11616. 57. A. Weltin, K. Slotwinski, J. Kieninger, I. Moser, G. Jobst, M.
29. J. Li, F. Rossignol and J. Macdonald, Lab Chip, 2015, 15, Wego, R. Ehretc and G. A. Urban, Lab Chip, 2014, 14, 138 -
2538 - 2558. 146.
Published on 13 February 2020. Downloaded on 2/15/2020 1:20:42 AM.

30. M. Song, L. Dang, J. Long and C. Hu, ACS Sens., 2018, 3, 58. J. Marzioch, J. Kieninger, A. Weltin, H. Flamm, K.
2518 - 2525. Aravindalochanan, J. A. Sandvik, E. O. Pettersen, Q. Peng
31. C. M. Fung, J. S. Lloyd, S. Samavat, D. Deganello and K. S. and G. A. Urban, Lab Chip, 2018, 18, 3353 - 3360.
Teng, Sens. Actuators B, 2017, 247, 807 - 813. 59. A. Yamagishi, K. Tanabe, M. Yokokawa, Y. Morimoto, M.
32. M. Bariya, Z. Shahpar, H. Park, J. Sun, Y. Jung, W. Gao, H. Kinoshita and H. Suzuki, Anal. Chim. Acta, 2017, 985, 1 - 6.
Y. Y. Nyein, T. S. Liaw, L.-C. Tai, Q. P. Ngo, M. Chao, Y. 60. C. E. Nwankire, A. Venkatanarayanan, T. Glennon, T. E.
Zhao, M. Hettick, G. Cho and A. Javey, ACS Nano, 2018, Keyes, R. J. Forster and J. Ducrée, Biosens. Bioelectron.,
12, 6978 - 6987. 2015, 68, 382 - 389.
33. J. Janata, Principles of chemical sensors, Springer, New 61. K. Sanger, K. Zór, C. B. Jendresen, A. Heiskanen, L. Amato,
York, 2 nd edn., 2009. A. T. Nielsen and A. Boisen, Sens. Actuators B, 2017, 253,
34. E. Zdrachek and E. Bakker, Anal. Chem., 2019, 91, 2 - 26. 999 - 1005
35. J. Hu, A. Stein and P. Bühlmann, Trends Anal. Chem., 2016, 62. T. Li, Y. Fan, Y. Cheng and J. Yang, Lab Chip, 2013, 13, 2634
76, 102 - 114. - 2640.
36. Handbook of reference electrodes, Springer, Heidelberg, 63. T.-H. Kim, K. Abi-Samra, V. Sunkara, D.-K. Park, M. Amasia,
2013. N. Kim, J. Kim, H. Kim, M. Madou and Y.-K. Cho, Lab Chip,
37. M. Sophocleous and J. K. Atkinson, Sens. Actuators A, 2013, 13, 3747 – 3754.
2017, 267, 106 - 120. 64. Y. Song, Y.-Y. Huang, X. Liu, X. Zhang, M. Ferrari and L.
38. M. Parrilla, M. Cuartero and G. A. Crespo, Trends Anal. Qin, Trends Biotechnol., 2014, 32, 132 - 139.
Chem., 2019, 110, 303 - 320. 65. M. Bauer, J. Bartoli, S. O. Martinez-Chapa and M. Madou,
39. E. Lindner, M. Guzinski, T. A. Khan and B. D. Pendley, ACS Micromachines, 2019, 10, 31.
Sens., 2019, 4, 549 - 561. 66. S. M. T. Delgado, J. G. Korvink and D. Mager, Biosens.
40. T. Kakiuchi, T. Yoshimatsu and N. Nishi, Anal. Chem., 2007, Bioelectron., 2018, 117, 464 - 473.
79, 7187 - 7191. 67. S. T. Rajendran, E. Scarano, M. H. Bergkamp, A. M. Capria,
41. L. Zhang, T. Miyazawa, Y. Kitazumi and T. Kakiuchi, Anal. C.-H. Cheng, K. Sanger, G. Ferrari, L. H. Nielsen, E.-T. Hwu,
Chem., 2012, 84, 3461 - 3464. K. Zór and A. Boisen, Anal. Chem., 2019, 91, 11620 -
42. S. A. Chopade, E. L. Anderson, P. W. Schmidt, T. P. Lodge, 11628.
M. A. Hillmyer and P. Bühlmann, ACS Sens., 2017, 2, 1498 68. M. D. M. Dryden, D. D. G. Rackus, M. H. Shamsi and A. R.
- 1504. Wheeler, Anal. Chem., 2013, 85, 8809 - 8816.
43. J. Horak, C. Dincer, H. Bakirci and G. Urban, Sens. 69. Y. Yu, J. Chen, J. Li, S. Yang, S.-K. Fan and J. Zhou, J.
Actuators B, 2014, 191, 813 - 820. Micromech. Microeng., 2013, 23, 095025.
44. A. Fragoso, D. Latta, N. Laboria, F. v. Germar, T. E. Hansen- 70. A. Farzbod and H. Moon, Biosens. Bioelectron., 2018, 106,
Hagge, W. Kemmner, C. Gärtner, R. Klemm, K. S. Dreseb 37 - 42.
and C. K. O’Sullivan, Lab Chip, 2011, 11, 625 - 631. 71. Y. Yu, J. Chen and J. Zhou, J. Micromech. Microeng., 2014,
45. G. Lee, I. Park, K. Kwon, T. Kwon, J. Seo, W.-J. Chang, H. 24, 015020.
Nam, G. S. Cha, M. H. Choi, D. S. Yoon and S. W. Lee, 72. M. H. Shamsi, K. Choi, A. H. C. Ng and A. R. Wheeler, Lab
Biomed Microdevices, 2012, 14, 375 - 384. Chip, 2014, 14, 547 - 554.
46. G. H. Lee, J. K. Lee, J. H. Kim, H. S. Choi, J. Kim, S. H. Lee 73. D. G. Rackus, M. D. M. Dryden, J. Lamanna, A. Zaragoza, B.
and H. Lee, Sci. Rep., 2017, 7, 7545. Lam, S. O. Kelley and A. R. Wheeler, Lab Chip, 2015, 15,
47. S. Kimura, J. Fukuda, A. Tajima and H. Suzuki, Lab Chip, 3776 - 3784.
2012, 12, 1309 - 1315. 74. A. Bratov, N. Abramova and A. Ipatov, Anal. Chim. Acta,
48. J. Kim, Y. Shin, S. Song, J. Lee and J. Kim, Sens. Actuators B, 2010, 678, 149 - 159.
2014, 202, 60 - 66.

26 | J. Name., 2012, 00, 1-3 This journal is © The Royal Society of Chemistry 20xx

Please do not adjust margins

 Page 27 of 32 Lab
Please do onadjust
not a Chipmargins
Journal Name
75. B. M. Lowe, K. Sun, I. Zeimpekis, C.-K. Skylaris and N. G.
Green, Analyst, 2017, 142, 4173 - 4200.
76. Y.-C. Syu, W.-E. Hsu and C.-T. Lin, ECS J. Solid State Sci.
Technol., 2018, 7, Q3196 - Q3207.
77. S. Szunerits and R. Boukherroub, Interface Focus, 2018, 8,
20160132.
78. A. Yamada and M. Suzuki, Sensors, 2017, 17, 1563.
79. S. Sharma, A. R.-B. Moniz, I. Triantis, K. Michelakis, J.
Trzebinski, A. Azarbadegan, B. Field, C. Toumazou, I.
Eames and A. Cass, Microfluid. Nanofluid., 2011, 10, 1119
- 1125.
80. K. Matsuura, Y. Asano, A. Yamada and K. Naruse, Sensors,
2013, 13, 2484 - 2493.
81. M. S. Andrianova, O. V. Gubanova, N. V. Komarova, E. V.
Kuznetsov and A. E. Kuznetsov, Electroanalysis, 2016, 28,
1311 - 1321.
82. A. E. Kuznetsov, N. V. Komarova, E. V. Kuznetsov, M. S.
Andrianova, V. P. Grudtsov, E. N. Rybachek, K. V. Puchnin,
D. V. Ryazantsev and A. N. Saurov, Biosens. Bioelectron.,
2019, 129, 29 - 35.
Published on 13 February 2020. Downloaded on 2/15/2020 1:20:42 AM.
83. Y.-H. Lin, S.-H. Wang, M.-H. Wu, T.-M. Pan, C.-S. Lai, J.-D.
Luo and C.-C. Chiou, Biosens. Bioelectron., 2013, 43, 328 -
335.
84. Y. Wang, Z. Ye, J. Ping, S. Jing and Y. Ying, Biosens.
Bioelectron., 2014, 59, 106 - 111.
85. J. Daprà, L. H. Lauridsen, A. T. Nielsen and N. Rozlosnik,
Biosens. Bioelectron., 2013, 43, 315 - 320.
86. M. A. Ali, P. R. Solanki, S. Srivastava, S. Singh, V. V.
Agrawal, R. John and B. D. Malhotra, ACS Appl. Mater.
Interfaces, 2015, 7, 5837 - 5846.
87. M. S. Chiriacò, E. Primiceri, A. G. Monteduro, A. Bove, S.
Leporatti, M. Capello, S. Ferri-Borgogno, R. Rinaldi, F.
Novelli and G. Maruccio, Lab Chip, 2013, 13, 730 - 734.
88. A. Lakey, Z. Ali, S. M. Scott, S. Chebil, H. Korri-Youssoufi, S.
Hunor, A. Ohlander, M. Kuphal and J. S. Marti, Biosens.
Bioelectron., 2019, 129, 147 - 154.
89. J. Paredes, S. Becerro and S. Arana, Sens. Actuators B,
2014, 195, 667 - 676.
90. H. Etayash, K. Jiang, T. Thundat and K. Kaur, Anal. Chem.,
2014, 86, 1693 - 1700.
91. R. Manczak, M. Fouet, R. Courson, P.-L. Fabre, A.
Montrose, J. Sudor, A.-M. Gué and K. Reybier, Sens.
Actuators B, 2016, 230, 825 - 831
92. L. An, G. Wang, Y. Han, T. Li, P. Jin and S. Liu, Lab Chip,
2018, 18, 335 - 342.
93. G. L. Damhorst, C. E. Smith, E. M. Salm, M. M. Sobieraj, H.
Ni, H. Kong and R. Bashir, Biomed. Microdevices, 2013, 15,
895 - 905.
94. C.-C. Wu and D.-J. Yang, Biosens. Bioelectron., 2013, 43,
348 - 354.
95. N.-C. Chen, C.-H. Chen, M.-K. Chen, L.-S. Jang and M.-H.
Wang, Sens. Actuators B, 2014, 190, 570 - 577.
96. H. Park, D. Kim and K.-S. Yun, Sens. Actuators B, 2010,
150, 167 - 173.
97. M. Kim, T. Jung, Y. Kim, C. Lee, K. Woo, J. H. Seol and S.
Yang, Biosens. Bioelectron., 2015, 74, 1011 - 1015.
98. R. Wang, Y. Xu, H. Liu, J. Peng, J. Irudayaraj and F. Cui,
Biomed. Microdevices, 2017, 19, 34.
99. B. d. Moral-Zamora, J. Punter-Villagrassa, A. M. Oliva-
Brañas, J. M. Álvarez-Azpeitia, J. Colomer-Farrarons, J.
Samitier, A. Homs-Corbera and P. L. Miribel-Català,
Electrophoresis, 2015, 36, 1130 - 1141.
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins
ARTICLE
100. A. Mansoorifar, A. Koklu and A. Beskok, View Anal. Chem.,
Article Online
2019, 91, 4140 - 4148. DOI: 10.1039/C9LC01112A
101. A. Mansoorifar, A. Koklu, S. Ma, G. V. Raj and A. Beskok,
Anal. Chem., 2018, 90, 4320 - 4327.
102. M. Nakano, R. Hamada, H. Takayama, Y. Shonishi, T.
Hisajima, L. Mao and J. Suehiro, Sens. Actuators B, 2012,
173, 676 - 681.
103. S. G. Dastider, S. Barizuddin, M. Dweik and M. Almasri,
RSC Adv., 2013, 3, 26297 - 26306.
104. R. Hamada, H. Takayama, Y. Shonishi, L. Mao, M. Nakano
and J. Suehiro, Sens. Actuators B, 2013, 181, 439 - 445.
105. C.-H. Chuang, Y.-W. Huang and Y.-T. Wu, Sensors, 2011,
11, 11021 - 11035.
106. K. Onishi, J. Enomoto, T. Araki, R. Takagi, H. Suzuki and J.
Lab on a Chip Accepted Manuscript
Fukuda, Analyst, 2018, 143, 396 - 399.
107. R. Takagi, J. Fukuda, K. Nagata, Y. Yawata, N. Nomura and
H. Suzuki, Analyst, 2013, 138, 1000 - 1003.
108. H. Ben-Yoav, P. H. Dykstra, W. E. Bentley and R. Ghodssi,
Biosens. Bioelectron., 2015, 64, 579 - 585.
109. D. Huh, Y.-s. Torisawa, G. A. Hamilton, H. J. Kim and D. E.
Ingber, Lab Chip, 2012, 12, 2156 - 2164.
110. E. W. Esch, A. Bahinski and D. Huh, Nat. Rev. Drug Discov.,
2015, 14, 248 - 260.
111. B. Zhang and M. Radisic, Lab Chip, 2017, 17, 2395 - 2420.
112. S. N. Bhatia and D. E. Ingber, Nat. Biotechnol., 2014, 32,
760 - 772.
113. A. Moya, M. Ortega-Ribera, X. Guimerà, E. Sowade, M.
Zea, X. Illa, E. Ramon, R. Villa, J. Gracia-Sancho and G.
Gabriel, Lab Chip, 2018, 18, 2023 - 2035.
114. D. Bavli, S. Prill, E. Ezra, G. Levy, M. Cohen, M. Vinken, J.
Vanfleteren, M. Jaeger and Y. Nahmias, Proc. Natl. Acad.
Sci. U. S. A., 2016, 113, E2231 - E2240.
115. S. R. Shin, Y. S. Zhang, D.-J. Kim, A. Manbohi, H. Avci, A.
Silvestri, J. Aleman, N. Hu, T. Kilic, W. Keung, M. Righi, P.
Assawes, H. A. Alhadrami, R. A. Li, M. R. Dokmeci and A.
Khademhosseini, Anal. Chem., 2016, 88, 10019 - 10027.
116. S. R. Shin, T. Kilic, Y. S. Zhang, H. Avci, N. Hu, D. Kim, C.
Branco, J. Aleman, S. Massa, A. Silvestri, J. Kang, A.
Desalvo, M. A. Hussaini, S.-K. Chae, A. Polini, N. Bhise, M.
A. Hussain, H. Lee, M. R. Dokmeci and A. Khademhosseini,
Adv. Sci., 2017, 4, 1600522.
117. Y. S. Zhang, J. Aleman, S. R. Shin, T. Kilic, D. Kim, S. A. M.
Shaegh, S. Massa, R. Riahi, S. Chae, N. Hu, H. Avci, W.
Zhang, A. Silvestri, A. S. Nezhad, A. Manbohi, F. D. Ferrari,
A. Polini, G. Calzone, N. Shaikh, P. Alerasool, E. Budina, J.
Kang, N. Bhise, J. Ribas, A. Pourmand, A. Skardal, T.
Shupe, C. E. Bishop, M. R. Dokmeci, A. Atala and A.
Khademhosseini, Proc. Natl. Acad. Sci. U. S. A., 2017, 114,
E2293 - E2302.
118. J. Kieninger, A. Weltin, H. Flamm and G. A. Urban, Lab
Chip, 2018, 18, 1274 - 1291.
119. J. R. Soucy, A. J. Bindas, A. N. Koppes and R. A. Koppes,
iScience, 2019, 21, 521 - 548.
120. W. Dungchai, O. Chailapakul and C. S. Henry, Anal. Chem.,
2009, 81, 5821 - 5826.
121. D. M. Cate, J. A. Adkins, J. Mettakoonpitak and C. S.
Henry, Anal. Chem., 2015, 87, 19 - 41.
122. J. Adkins, K. Boehle and C. Henry, Electrophoresis, 2015,
36, 1811 - 1824.
123. Y. Lin, D. Gritsenko, S. Feng, Y. C. Teh, X. Lu and J. Xu,
Biosens. Bioelectron., 2016, 83, 256 - 266.
J. Name., 2013, 00, 1-3 | 27
 Lab
Please do onadjust
not a Chipmargins Page 28 of 32

ARTICLE Journal Name

124. J. Mettakoonpitak, K. Boehle, S. Nantaphol, P. Teengam, J. 150. F. Arduini, S. Cinti, V. Caratelli, L. Amendola, G. Palleschi
View Article Online
A. Adkins, M. Srisa-Art and C. S. Henry, Electroanalysis, and D. Moscone, Biosens. Bioelectron., 2019, 126, 346 -
DOI: 10.1039/C9LC01112A
2016, 28, 1420 - 1436. 354.
125. S. Ahmed, M.-P. N. Bui and A. Abbas, Biosens. 151. A. Yakoh, S. Chaiyo, W. Siangproh and O. Chailapakul, ACS
Bioelectron., 2016, 77, 249 - 263. Sens., 2019, 4, 1211 - 1221.
126. Y. Xia, J. Si and Z. Li, Biosens. Bioelectron., 2016, 77, 774 - 152. J. R. Windmiller and J. Wang, Electroanalysis, 2012, 24, 1 -
789. 18.
127. T. Akyazi, L. Basabe-Desmonts and F. Benito-Lopez, Anal. 153. J. R. Windmiller and J. Wang, Electroanalysis, 2013, 25, 29
Chim. Acta, 2018, 1001, 1 - 17. - 46.
128. V. B. C. Lee, N. F. Mohd-Naim, E. Tamiya and M. U. 154. A. J. Bandodkar and J. Wang, Trends Biotechnol., 2014, 32,
Ahmed, Anal. Sci., 2018, 34, 7 - 18. 363 - 371.
129. C. M. Silveira, T. Monteiro and M. G. Almeida, Biosensors, 155. G. Matzeu, L. Florea and D. Diamond, Sens. Actuators B,
2016, 6, 51. 2015, 211, 403 - 418.
130. C. Hu, X. Bai, Y. Wang, W. Jin, X. Zhang and S. Hu, Anal. 156. M. D. Steinberg, P. Kassal and I. M. Steinberg,

Lab on a Chip Accepted Manuscript

Chem., 2012, 84, 3745 - 3750. Electroanalysis, 2016, 28, 1149 - 1169.
131. A. Määttänen, U. Vanamo, P. Ihalainen, P. Pulkkinen, H. 157. K. Mitsubayashi and T. Arakawa, Electroanalysis, 2016, 28,
Tenhu, J. Bobacka and J. Peltonen, Sens. Actuators B, 1170 - 1187.
2013, 177, 153 - 162. 158. A. J. Bandodkar, I. Jeerapan and J. Wang, ACS Sens., 2016,
132. T. H. d. Costa, E. Song, R. P. Tortorich and J.-W. Choi, ECS 1, 464 - 482.
J. Solid State Sci. Technol., 2015, 4, S3044 - S3047. 159. Y. Liu, M. Pharr and G. A. Salvatore, ACS Nano, 2017, 11,
Published on 13 February 2020. Downloaded on 2/15/2020 1:20:42 AM.

133. M. Santhiago, C. C. Corrêa, J. S. Bernardes, M. P. Pereira, 9614 − 9635.

L. J. M. Oliveira, M. Strauss and C. C. B. Bufon, ACS Appl.
Mater. Interfaces, 2017, 9, 24365 - 24372.
134. C. L. S. Chagas, L. C. Duarte, E. O. Lobo-Júnior, E. Piccin, N.
Dossi and W. K. T. Coltro, Electrophoresis, 2015, 36, 1837 -
1844.
135. N. Dossi, R. Toniolo, F. Terzi, E. Piccin and G. Bontempelli,
Electrophoresis, 2015, 36, 1830 - 1836.
136. A. J. Bandodkar, W. Jia, J. Ramírez and J. Wang, Adv.
Healthcare Mater., 2015, 4, 1215 - 1224.
137. Z. Li, F. Li, Y. Xing, Z. Liu, M. You, Y. Li, T. Wen, Z. Qu, X. L.
Li and F. Xu, Biosens. Bioelectron., 2017, 98, 478 - 485.
138. R. Kawahara, P. Sahatiya, S. Badhulika and S. Uno, Jpn. J.
Appl. Phys., 2018, 57, 04FM08.
139. A. A. Dias, T. M. G. Cardoso, C. L. S. Chagas, V. X. G.
Oliveira, R. A. A. Munoz, C. S. Henry, M. H. P. Santana, T.
R. L. C. Paixão and W. K. T. Coltro, Electroanalysis, 2018,
30, 2250 - 2257.
140. V. Soum, Y. Kim, S. Park, M. Chuong, S. R. Ryu, S. H. Lee,
G. Tanev, J. Madsen, O.-S. Kwon and K. Shin,
Micromachines, 2019, 10, 109.
141. W. R. d. Araujo, C. M. R. Frasson, W. A. Ameku, J. R. Silva,
L. Angnes and T. R. L. C. Paixão, Angew. Chem. Int. Ed.,
2017, 56, 15113 - 15117.
142. X. Zang, C. Shen, Y. Chu, B. Li, M. Wei, J. Zhong, M.
Sanghadasa and L. Lin, Adv. Mater., 2018, 30, 1800062.
143. S. E. Fosdick, M. J. Anderson, C. Renault, P. R. DeGregory,
J. A. Loussaert and R. M. Crooks, Anal. Chem., 2014, 86,
3659 - 3666.
144. W. Li, L. Li, M. Li, J. Yu, S. Ge, M. Yan and X. Song, Chem.
Commun., 2013, 49, 9540 - 9542.
145. H. Liu, Y. Xiang, Y. Lu and R. M. Crooks, Angew. Chem. Int.
Ed., 2012, 51, 6925 - 6928.
146. P. Wang, L. Ge, M. Yan, X. Song, S. Ge and J. Yu, Biosens.
Bioelectron., 2012, 32, 238 - 243.
147. C. Ma, W. Li, Q. Kong, H. Yang, Z. Bian, X. Song, J. Yu and
M. Yan, Biosens. Bioelectron., 2015, 63, 7 - 13.
148. C.-C. Wang, J. W. Hennek, A. Ainla, A. A. Kumar, W.-J. Lan,
J. Im, B. S. Smith, M. Zhao and G. M. Whitesides, Anal.
Chem., 2016, 88, 6326 − 6333.
149. J. Ding, B. Li, L. Chen and W. Qin, Angew. Chem. Int. Ed.,
2016, 55, 13033 - 13037.
160. J. Choi, R. Ghaffari, L. B. Baker and J. A. Rogers, Sci. Adv.,
2018, 4, eaar3921.
161. J. Heikenfeld, A. Jajack, J. Rogers, P. Gutruf, L. Tian, T. Pan,
R. Li, M. Khine, J. Kim, J. Wang and J. Kim, Lab Chip, 2018,
18, 217 - 248.
162. P. Kassal, M. D. Steinberg and I. M. Steinberg, Sens.
Actuators B, 2018, 266, 228 - 245.
163. Y. Yang and W. Gao, Chem. Soc. Rev., 2019, 48, 1465 -
1491.
164. A. J. Bandodkar, R. Nuñez-Flores, W. Jia and J. Wang, Adv.
Mater., 2015, 27, 3060 - 3065.
165. A. J. Bandodkar, I. Jeerapan, J.-M. You, R. Nuũez-Flores
and J. Wang, Nano Lett., 2016, 16, 721 - 727.
166. H. Lee, T. K. Choi, Y. B. Lee, H. R. Cho, R. Ghaffari, L. Wang,
H. J. Choi, T. D. Chung, N. Lu, T. Hyeon, S. H. Choi and D.-
H. Kim, Nat. Nanotechnol., 2016, 11, 566 - 572.
167. J. R. Windmiller, A. J. Bandodkar, G. Valdés-Ramírez, S.
Parkhomovsky, A. G. Martinez and J. Wang, Chem.
Commun., 2012, 48, 6794 - 6796.
168. W. Jia, A. J. Bandodkar, G. Valdés-Ramírez, J. R.
Windmiller, Z. Yang, J. Ramírez, G. Chan and J. Wang,
Anal. Chem., 2013, 85, 6553 - 6560.
169. A. J. Bandodkar, V. W. S. Hung, W. Jia, G. Valdés-Ramírez,
J. R. Windmiller, A. G. Martinez, J. Ramírez, G. Chan, K.
Kerman and J. Wang, Analyst, 2013, 138, 123 - 128.
170. J. Kim, W. R. d. Araujo, I. A. Samek, A. J. Bandodkar, W.
Jia, B. Brunetti, T. R. L. C. Paixão and JosephWang,
Electrochem. Commun., 2015, 51, 41 - 45.
171. T. Guinovart, A. J. Bandodkar, J. R. Windmiller, F. J.
Andrade and J. Wang, Analyst, 2013, 138, 7031 - 7038.
172. D. Pankratov, E. González-Arribas, Z. Blum and S. Shleev,
Electroanalysis, 2016, 28, 1250 - 1266.
173. H. Yao, Y. Liao, A. R. Lingley, A. Afanasiev, I. Lähdesmäki,
B. P. Otis and B. A. Parviz, J. Micromech. Microeng., 2012,
22, 075007.
174. H. Yao, A. J. Shum, M. Cowan, I. Lähdesmäki and B. A.
Parviz, Biosens. Bioelectron., 2011, 26, 3290 - 3296.
175. T. Choudhary, G. P. Rajamanickam and D. Dendukuri, Lab
Chip, 2015, 15, 2064 - 2072.
176. X. Liu and P. B. Lillehoj, Lab Chip, 2016, 16, 2093 - 2098.

28 | J. Name., 2012, 00, 1-3 This journal is © The Royal Society of Chemistry 20xx

Please do not adjust margins

 Page 29 of 32 Lab
Please do onadjust
not a Chipmargins
Journal Name
177. K. Nagamine, T. Mano, A. Nomura, Y. Ichimura, R. Izawa,
H. Furusawa, H. Matsui, D. Kumaki and S. Tokito, Sci. Rep.,
2019, 9, 10102.
178. S. Lin, B. Wang, Y. Zhao, R. Shih, X. Cheng, W. Yu, H.
Hojaiji, H. Lin, C. Hoffman, D. Ly, J. Tan, Y. Chen, D. D.
Carlo, C. Milla and S. Emaminejad, ACS Sens., 2020, 5, 93 -
102.
179. J. Gonzalo-Ruiz, R. Mas, C. d. Haro, E. Cabruja, R. Camero,
M. A. Alonso-Lomillo and F. J. Muñoz, Biosens.
Bioelectron., 2009, 24, 1788 - 1791.
180. J. Kim, I. Jeerapan, S. Imani, T. N. Cho, A. Bandodkar, S.
Cinti, P. P. Mercier and J. Wang, ACS Sens., 2016, 1, 1011 -
1019.
181. S. Emaminejad, W. Gao, E. Wu, Z. A. Davies, H. Y. Y. Nyein,
S. Challa, S. P. Ryan, H. M. Fahad, K. Chen, Z. Shahpar, S.
Talebi, C. Milla, A. Javey and R. W. Davis, Proc. Natl. Acad.
Sci. U. S. A., 2017, 114, 4625 - 4630.
182. J. Kim, J. R. Sempionatto, S. Imani, M. C. Hartel, A.
Barfidokht, G. Tang, A. S. Campbell, P. P. Mercier and J.
Wang, Adv. Sci., 2018, 5, 1800880.
Published on 13 February 2020. Downloaded on 2/15/2020 1:20:42 AM.
183. P. R. Miller, X. Xiao, I. Brener, D. B. Burckel, R. Narayan
and R. Polsky, Adv. Healthcare Mater., 2014, 3, 876 - 881.
184. P. Miller, M. Moorman, R. Manginell, C. Ashlee, I. Brener,
D. Wheeler, R. Narayan and R. Polsky, Electroanalysis,
2016, 28, 1305 - 1310.
185. A. Jina, M. J. Tierney, J. A. Tamada, S. McGill, S. Desai, B.
Chua, A. Chang and M. Christiansen, J. Diabetes Sci.
Technol., 2014, 8, 483 - 487.
186. J. R. Windmiller, G. Valdés-Ramírez, N. Zhou, M. Zhou, P.
R. Miller, C. Jin, S. M. Brozik, R. Polsky, E. Katz, R. Narayan
and J. Wang, Electroanalysis, 2011, 23, 2302 - 2309.
187. A. Caliò, P. Dardano, V. D. Palma, M. F. Bevilacqua, A. D.
Matteo, H. Iueleb and L. D. Stefano, Sens. Actuators B,
2016, 236, 343 - 349.
188. P. Bollella, S. Sharma, A. E. G. Cass and R. Antiochia,
Biosens. Bioelectron., 2019, 123, 152 - 159.
189. P. Bollella, S. Sharma, A. E. G. Cass and R. Antiochia,
Electroanalysis, 2019, 31, 374 - 382.
190. A. J. Bandodkar, D. Molinnus, O. Mirza, T. Guinovart, J. R.
Windmiller, G. Valdés-Ramírez, F. J. Andrade, M. J.
Schöning and J. Wang, Biosens. Bioelectron., 2014, 54, 603
- 609.
191. J. Kim, S. Imani, W. R. d. Araujo, J. Warchall, G. Valdés-
Ramírez, T. R. L. C. Paixão, P. P. Mercier and J. Wang,
Biosens. Bioelectron., 2015, 74, 1061 - 1068.
192. A. Martín, J. Kim, J. F. Kurniawan, J. R. Sempionatto, J. R.
Moreto, G. Tang, A. S. Campbell, A. Shin, M. Y. Lee, X. Liu
and J. Wang, ACS Sens., 2017, 2, 1860 - 1868.
193. T. Arakawa, Y. Kuroki, H. Nitta, P. Chouhan, K. Toma, S.-i.
Sawada, S. Takeuchi, T. Sekita, K. Akiyoshi, S. Minakuchi
and K. Mitsubayashi, Biosens. Bioelectron., 2016, 84, 106 -
111.
194. Y.-T. Liao, H. Yao, A. Lingley, B. Parviz and B. P. Otis, IEEE J.
Solid-State Circuit, 2012, 47, 335 - 344.
195. C. M. Caffrey, K. Twomey and V. I. Ogurtsov, Sens.
Actuators B, 2015, 218, 8 - 15.
196. R. K. Mishra, L. J. Hubble, A. Martín, R. Kumar, A.
Barfidokht, J. Kim, M. M. Musameh, I. L. Kyratzis and J.
Wang, ACS Sens., 2017, 2, 553 - 561.
197. D. Zhang and Q. Liu, Biosens. Bioelectron., 2016, 75, 273 -
284.
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins
ARTICLE
198. D. Xu, X. Huang, J. Guo and X. Ma, Biosens.View Bioelectron.,
Article Online
2018, 110, 78 - 88. DOI: 10.1039/C9LC01112A
199. X. Huang, D. Xu, J. Chen, J. Liu, Y. Li, J. Song, X. Ma and J.
Guo, Analyst, 2018, 143, 5339 - 5351.
200. J. Liu, Z. Geng, Z. Fan, J. Liu and H. Chen, Biosens.
Bioelectron., 2019, 132, 17 - 37.
201. A. C. Sun and D. A. Hall, Electroanalysis, 2019, 31, 2 - 16.
202. P. Gravesen, J. Branebjerg and O. S. Jensen, J. Micromech.
Microeng., 1993, 3, 168 - 182.
203. S. Shoji and M. Esashi, J. Micromech. Microeng., 1994, 4,
157 - 171.
204. D. J. Laser and J. G. Santiago, J. Micromech. Microeng.,
2004, 14, R35 - R64.
205. C. Zhang, D. Xing and Y. Li, Biotechnol. Adv., 2007, 25, 483
Lab on a Chip Accepted Manuscript
- 514.
206. S. Böhm, B. Timmer, W. Olthuis and P. Bergveld, J.
Micromech. Microeng., 2000, 10, 498-504.
207. H. Gensler, R. Sheybani, P.-Y. Li, R. Lo and E. Meng,
Biomed Microdevices., 2012, 14, 483 - 496.
208. Y. Yi, U. Buttner, A. A. A. Carreno, D. Conchouso and I. G.
Foulds, J. Micromech. Microeng., 2015, 25, 105011.
209. X. Li, D. Li, X. Liu and H. Chang, Sens. Actuators B, 2016,
229, 466 - 475.
210. H. Kim, H. Hwang, S. Baek and D. Kim, Sens. Actuators A,
2018, 277, 73 - 84.
211. J. Nestler, A. Morschhauser, K. Hiller, T. Otto, S. Bigot, J.
Auerswald, H. F. Knapp, J. Gavillet and T. Gessner, Int. J.
Adv. Manuf. Technol., 2010, 47, 137 - 145.
212. H. Obata, T. Kuji, K. Kojima, F. Sassa, M. Yokokawa, K.
Takekosh and H. Suzuki, ACS Sens., 2016, 1, 190–196.
213. G. Blanco-Gomez, A. Glidle, L. M. Flendrig and J. M.
Cooper, Anal. Chem., 2009, 81, 1365 - 1370.
214. S. Schumacher, J. Nestler, T. Otto, M. Wegener, E.
Ehrentreich-Förster, D. Michel, K. Wunderlich, S. Palzer, K.
Sohn, A. Weber, M. Burgard, A. Grzesiak, A. Teichert, A.
Brandenburg, B. Koger, J. Albers, E. Nebling and F. F. Bier,
Lab Chip, 2012, 12, 464 - 473.
215. M. L. Y. Sin, V. Gau, J. C. Liao and P. K. Wong, J
Microelectromech. Syst., 2013, 22, 1126 - 1132.
216. A. Chen, R. Wang, C. R. S. Bever, S. Xing, B. D. Hammock
and T. Pan, Biomicrofluidics, 2014, 8, 064101.
217. S. Geidel, S. P. Llopis, M. Rodrigo, G. d. Diego-Castilla, A.
Sousa, J. Nestler, T. Otto, T. Gessner and V. Parro,
Micromachines, 2016, 7, 153.
218. E. Delamarche, D. Juncker and H. Schmid, Adv. Mater.,
2005, 17, 2911-2933.
219. W. Satoh, H. Yokomaku, H. Hosono, N. Ohnishi and H.
Suzuki, J. Appl. Phys., 2008, 103, 034903.
220. W. Satoh, S. Takahashi, F. Sassa, J. Fukuda and H. Suzuki,
Lab Chip, 2009, 9, 35 - 37.
221. G. Biswas, T. Watanabe, E. T. Carlen, M. Yokokawa and H.
Suzuki, ChemPhysChem, 2016, 17, 817 - 821.
222. S. Yamaguchi, K. Morimoto, J. Fukuda and H. Suzuki,
Biosens. Bioelectron., 2009, 24, 2171 - 2176.
223. J. Chen, Y. Zhou, D. Wang, F. He, V. M. Rotello, K. R.
Carter, J. J. Watkins and S. R. Nugen, Lab Chip, 2015, 15,
3086 - 3094.
224. A. Ezkerra, L. J. Fernández, K. Mayora and J. M. Ruano-
López, Sens. Actuators B, 2011, 155, 505 - 511.
225. P. Siribunbandal, S. Yamaguchi, K. Kojima, J. Fukuda and
H. Suzuki, Appl. Phys. Lett., 2009, 95, 204105.
J. Name., 2013, 00, 1-3 | 29
 Lab
Please do onadjust
not a Chipmargins Page 30 of 32

ARTICLE Journal Name

226. T. Watanabe, G. C. Biswas, E. T. Carlen and H. Suzuki, RSC 256. Z. Farka, T. Juřík, D. Kovář, L. Trnková and P. Skládal,
View Article Online
Adv., 2017, 7, 39018 - 39023. Chem. Rev., 2017, 117, 9973 - 10042. DOI: 10.1039/C9LC01112A
227. D. Welch and J. B. Christen, Lab Chip, 2014, 14, 1191 - 257. N. Wongkaew, M. Simsek, C. Griesche and A. J. Baeumner,
1197. Chem. Rev., 2019, 119, 120 - 194.
228. K. Morimoto, M. Toya, J. Fukuda and H. Suzuki, Anal. 258. J. F. Rusling, Anal. Chem., 2013, 85, 5304 - 5310.
Chem., 2008, 80, 905 - 914. 259. M.-C. Chuang, H.-Y. Lai, J.-a. A. Ho and Y.-Y. Chen, Biosens.
229. S. O. Krabbenborg, C. Nicosia, P. Chen and J. Huskens, Bioelectron., 2013, 41, 602 - 607.
Nat. Commun., 2013, 4, 1667. 260. F. Picollo, A. Battiato, E. Bernardi, A. Marcantoni, A.
230. H. Zhou, G. Li and S. Yao, Lab Chip, 2014, 14, 1917 - 1922. Pasquarelli, E. Carbone, P. Olivero and V. Carabelli, Anal.
231. K. Štulík, C. Amatore, K. Holub, V. Mareček and W. Kutner, Chem., 2016, 88, 7493 - 7499.
Pure Appl. Chem., 2000, 72, 1483 - 1492. 261. J. Wang, R. Trouillon, Y. Lin, M. I. Svensson and A. G.
232. S. Carroll, M. M. Marei, T. J. Roussel, R. S. Keynton and R. Ewing, Anal. Chem. , 2013, 85, 5600 - 5608.
P. Baldwin, Sens. Actuators B, 2011, 160, 318 - 326. 262. J. Wang, R. Trouillon, J. Dunevall and A. G. Ewing, Anal.
233. F. Sassa, H. Laghzali, J. Fukuda and H. Suzuki, Anal. Chem., Chem., 2014, 86, 4515 - 4520.

Lab on a Chip Accepted Manuscript

2010, 82, 8725 - 8732. 263. A. Yakushenko, E. Kätelhön and B. Wolfrum, Anal. Chem.,
234. K. Ikemoto, T. Seki, S. Kimura, Y. Nakaoka, S. Tsuchiya, F. 2013, 85, 5483 - 5490.
Sassa, M. Yokokawa and H. Suzuki, Anal. Chem. , 2016, 88, 264. K. Kisler, B. N. Kim, X. Liu, K. Berberian, Q. Fang, C. J.
9427 - 9434. Mathai, S. Gangopadhyay, K. D. Gillis and M. Lindau, J.
235. I. Anshori, S. Takie and H. Suzuki, Electroanalysis, 2019, Biomater. Nanobiotechnol., 2012, 3, 243 - 253.
31, 1630 - 1634. 265. V. Carabelli, S. Gosso, A. Marcantoni, Y. Xu, E. Colombo, Z.
Published on 13 February 2020. Downloaded on 2/15/2020 1:20:42 AM.

236. O. Niwa, Electroanalysis, 1995, 7, 606 - 613. Gao, E. Vittone, E. Kohn, A. Pasquarelli and E. Carbone,
237. K. Hayashi, J.-i. Takahashi, T. Horiuchi, Y. Iwasaki and T. Biosens. Bioelectron., 2010, 26, 92 - 98.
Haga, J. Electrochem. Soc., 2008, 155, J240 - J243. 266. J. Wigström, J. Dunevall, N. Najafinobar, J. Lovrić, J. Wang,
238. S. Partel, C. Dincer, S. Kasemann, J. Kieninger, J. Edlinger A. G. Ewing and A.-S. Cans, Anal. Chem., 2016, 88, 2080 -
and G. Urban, ACS Nano, 2016, 10, 1086 - 1092. 2087.
239. E. D. Goluch, B. Wolfrum, P. S. Singh, M. A. G. 267. C. Amatore, J. Delacotte, M. Guille-Collignon and F.
Zevenbergen and S. G. Lemay, Anal. Bioanal. Chem., 2009, Lemaître, Analyst, 2015, 140, 3687 - 3695.
394, 447 - 456. 268. J. D. Besant, E. H. Sargent and S. O. Kelley, Lab Chip, 2015,
240. E. Kätelhön, B. Hofmann, S. G. Lemay, M. A. G. 15, 2799 - 2807.
Zevenbergen, A. Offenhäusser and B. Wolfrum, Anal. 269. A. Steude, S. Schmidt, A. A. Robitzki and O. Pänke, Lab
Chem., 2010, 82, 8502 - 8509. Chip, 2011, 11, 2884 - 2892.
241. L. Rassaei, K. Mathwig, S. Kang, H. A. Heering and S. G. 270. S. Abdellaoui, A. Noiriel, R. Henkens, C. Bonaventura, L. J.
Lemay, ACS Nano, 2014, 8 8278 - 8284. Blum and B. Doumèche, Anal. Chem., 2013, 85, 3690 -
242. D. Han, I. Lawrence P. Zaino, K. Fu and P. W. Bohn, J. Phys. 3697.
Chem. C, 2016, 120, 20634 - 20641. 271. Z. Wu, C.-H. Zhou, L.-J. Pan, T. Zeng, L. Zhu, D.-W. Pang
243. M. Atighilorestani and A. G. Brolo, Anal. Chem., 2017, 89, and Z.-L. Zhang, Anal. Chem., 2016, 88, 9166 - 9172.
9870 - 9876. 272. X. Luo, Q. Xu, T. James and J. J. Davis, Anal. Chem., 2014,
244. C. Ma, N. M. Contento, I. Larry R. Gibson and P. W. Bohn, 86, 5553 - 5558.
Anal. Chem., 2013, 85, 9882 - 9888. 273. A. Kling, C. Chatelle, L. Armbrecht, E. Qelibari, J. Kieninger,
245. C. Ma, N. M. Contento and P. W. Bohn, J. Am. Chem. Soc., C. Dincer, W. Weber and G. Urban, Anal. Chem., 2016, 88,
2014, 136, 7225 - 7228. 10036 - 10043.
246. Y. Kanno, K. Ino, H. Shikua and T. Matsue, Lab Chip, 2015, 274. C. K. Tang, A. Vaze and J. F. Rusling, Lab Chip, 2012, 12,
15, 4404 - 4414. 281 - 286.
247. F. Zhu, J. Yan, S. Pang, Y. Zhou, B. Mao, A. Oleinick, I. Svir 275. B. A. Otieno, C. E. Krause, A. Latus, B. V. Chikkaveeraiah,
and C. Amatore, Anal. Chem., 2014, 86, 3138 - 3145. R. C. Faria and J. F. Rusling, Biosens. Bioelectron., 2014,
248. C. Ma, L. P. Z. III and P. W. Bohn, Chem. Sci., 2015, 6, 3173 53, 268 - 274.
- 3179. 276. B. A. Otieno, C. E. Krause, A. L. Jones, R. B. Kremer and J.
249. S. Park, J. H. Park, S. Hwang and J. Kwak, Electrochem. F. Rusling, Anal. Chem., 2016, 88, 9269 - 9275.
Commun., 2016, 68, 76 - 80. 277. R. Malhotra, V. Patel, B. V. Chikkaveeraiah, B. S. Munge, S.
250. N. Y. Adly, B. Bachmann, K. J. Krause, A. Offenhäusser, B. C. Cheong, R. B. Zain, M. T. Abraham, D. K. Dey, J. S.
Wolfrumab and A. Yakushenko, RSC Adv., 2017, 7, 5473 - Gutkind and J. F. Rusling, Anal. Chem., 2012, 84, 6249 -
5479. 6255.
251. M. Holzinger, A. L. Goff and S. Cosnier, Front. Chem., 278. C. K. Tang, A. Vaze, M. Shen and J. F. Rusling, ACS Sens.,
2014, 2, 63. 2016, 1, 1036 - 1043.
252. C. Yang, M. E. Denno, P. Pyakurel and B. J. Venton, Anal. 279. J. P. Pursey, Y. Chen, E. Stulz, M. K. Park and P.
Chim. Acta, 2015, 887, 17 - 37. Kongsuphol, Sens. Actuators B, 2017, 251, 34 - 39.
253. C. Zhu, G. Yang, H. Li, D. Du and Y. Lin, Anal. Chem., 2015, 280. J. D. Slinker, N. B. Muren, A. A. Gorodetsky and J. K.
87, 230 - 249. Barton, J. Am. Chem. Soc., 2010, 132, 2769 - 2774.
254. J. N. Tiwari, V. Vij, K. C. Kemp and K. S. Kim, ACS Nano, 281. K. Ino, W. Saito, M. Koide, T. Umemura, H. Shiku and T.
2016, 10, 46 - 80. Matsue, Lab Chip, 2011, 11, 385 - 388.
255. G. T. Chandran, X. Li, A. Ogata and R. M. Penner, Anal. 282. K. Ino, T. Nishijo, T. Arai, Y. Kanno, Y. Takahashi, H. Shiku
Chem., 2017, 89, 249 - 275.

30 | J. Name., 2012, 00, 1-3 This journal is © The Royal Society of Chemistry 20xx

Please do not adjust margins

 Page 31 of 32 Lab
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not a Chipmargins
Journal Name
and T. Matsue, Angew. Chem. Int. Ed., 2012, 51, 6648 -
6652.
283. M. Şen, K. Ino, H. Shiku and T. Matsue, Lab Chip, 2012, 12,
4328 - 4335.
284. E. Kätelhön, D. Mayer, M. Banzet, A. Offenhäusser and B.
Wolfrum, Beilstein J. Nanotechnol., 2014, 5, 1137 - 1143.
285. B. Lam, J. Das, R. D. Holmes, L. Live, A. Sage, E. H. Sargent
and S. O. Kelley, Nat. Commun., 2013, 4, 2001.
286. H. Zhang, T. Oellers, W. Feng, T. Abdulazim, E. N. Saw, A.
Ludwig, P. A. Levkin and N. Plumeré, Anal. Chem., 2017,
89, 5832 - 5839.
287. F. Mavré, K.-F. Chow, E. Sheridan, B.-Y. Chang, J. A. Crooks
and R. M. Crooks, Anal. Chem., 2009, 81, 6218 - 6225.
288. F. Mavré, R. K. Anand, D. R. Laws, K.-F. Chow, B.-Y. Chang,
J. A. Crooks and R. M. Crooks, Anal. Chem., 2010, 82, 8766
- 8774.
289. S. E. Fosdick, K. N. Knust, K. Scida and R. M. Crooks,
Angew. Chem. Int. Ed., 2013, 52, 10438 - 10456.
290. X. Zhang, Q. Zhai, H. Xing, J. Li and E. Wang, ACS Sens.,
2017, 2, 320 - 326.
Published on 13 February 2020. Downloaded on 2/15/2020 1:20:42 AM.
291. R. M. Crooks, ChemElectroChem, 2016, 3, 357 - 359.
292. Y. Xiao, L. Xu and L.-W. Qi, Talanta, 2017, 165, 577 - 583.
293. K.-F. Chow, F. Mavré, J. A. Crooks, B.-Y. Chang and R. M.
Crooks, J. Am. Chem. Soc., 2009, 131, 8364 - 8365.
294. X. Zhang, C. Chen, J. Li, L. Zhang and E. Wang, Anal. Chem.,
2013, 85, 5335 - 5339.
295. W. W. Ye, J. Y. Shi, C. Y. Chan, Y. Zhang and M. Yang, Sens.
Actuators B, 2014, 193, 877 - 882.
296. J. P. Guerrette, S. J. Percival and B. Zhang, J. Am. Chem.
Soc., 2013, 135, 855 - 861.
297. S. M. Oja and B. Zhang, Anal. Chem., 2014, 86, 12299 -
12307.
298. M.-S. Wu, Z. Liu, H.-W. Shi, H.-Y. Chen and J.-J. Xu, Anal.
Chem., 2015, 87, 530 - 537.
299. Y.-Z. Wang, C.-H. Xu, W. Zhao, Q.-Y. Guan, H.-Y. Chen and
J.-J. Xu, Anal. Chem., 2017, 89, 8050 - 8056.
300. K.-F. Chow, F. Mavré and R. M. Crooks, J. Am. Chem. Soc.,
2008, 130, 7544 - 7545.
301. J.-D. Zhang, W.-W. Zhao, J.-J. Xu and H.-Y. Chen, J.
Electroanal. Chem., 2016, 781, 56 - 61.
302. K. Y. Inoue, M. Ikegawa, T. Ito-Sasaki, S. Takano, H. Shiku
and T. Matsue, ChemElectroChem, 2018, 5, 2167 - 2170.
303. D. G. Hafeman, J. W. Parce and H. M. McConnell, Science,
1988, 240, 1182 - 1185.
304. F. Wu, I. Campos, D.-W. Zhang and S. Krause, Proc. R. Soc.
A, 2017, 473, 20170130.
305. J. Wang, L. Du, S. Krause, C. Wu and P. Wang, Sens.
Actuators B, 2018, 265, 161 - 173.
306. C. Wu, T. Bronder, A. Poghossian, C. F. Werner, M. Bäcker
and M. J. Schöning, Phys. Status Solidi A, 2014, 211, 1423 -
1428.
307. S. Takenaga, C. Herrera, C. F. Werner, M. Biselli, V.
Thönnessen, T. Schnitzler, P. Öhlschläger, F. N. Almajhdi,
T. Wagner and M. J. Schöning, Phys. Status Solidi A, 2014,
211, 1410 - 1415.
308. L. Du, C. Wu, H. Peng, L. Zhao, L. Huang and P. Wang,
Biosens. Bioelectron., 2013, 40, 401 - 406.
309. K.-i. Miyamoto, A. Itabashi, T. Wagner, M. J. Schöning and
T. Yoshinobu, Sens. Actuators B, 2014, 194, 521 - 527.
310. W. Hu, H. Cai, J. Fu, P. Wang and G. Yang, Sens. Actuators
B, 2008, 129, 397 - 403.
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins
ARTICLE
311. T. Wagner, C. F. Werner, K.-i. Miyamoto, M. J. Schöning
View Article Online
and T. Yoshinobu, Sens. Actuators B, 2012,
DOI: 170, 34 - 39.
10.1039/C9LC01112A
312. TorstenWagner, W. Vornholt, C. FrederikWerner, T.
Yoshinobu, K.-i. Miyamoto, M. Keusgen and M. J.
Schöning, Phys. Med., 2016, 1, 2 - 7.
313. P. M. Shaibani, H. Etayash, S. Naicker, K. Kaur and T.
Thundat, ACS Sens., 2017, 2, 151 - 156.
314. P. M. Shaibani, H. Etayash, K. Jiang, A. Sohrabi, M.
Hassanpourfard, S. Naicker, M. Sadrzadeh and T. Thundat,
ACS Sens., 2018, 3, 815 - 822.
315. N. Hu, C. Wu, D. Ha, T. Wang, Q. Liu and P. Wang, Biosens.
Bioelectron., 2013, 40, 167 - 173.
316. Y. B. Vogel, J. J. Gooding and S. Ciampi, Chem. Soc. Rev.,
2019, 48, 3723 - 3739.
Lab on a Chip Accepted Manuscript
317. M. H. Choudhury, S. Ciampi, Y. Yang, R. Tavallaie, Y. Zhu, L.
Zarei, V. R. Gonçales and J. J. Gooding, Chem. Sci., 2015, 6,
6769 - 6776.
318. D. Seo, S. Y. Lim, J. Lee, J. Yun and T. D. Chung, ACS Appl.
Mater. Interfaces, 2018, 10, 33662 - 33668.
319. S. Joo and R. B. Brown, Chem. Rev., 2008, 108, 638 - 651.
320. S. K. Arya, C. C. Wong, Y. J. Jeon, T. Bansal and M. K. Park,
Chem. Rev., 2015, 115, 5116 - 5158.
321. B. N. Kim, A. D. Herbst, S. J. Kim, B. A. Minch and M.
Lindau, Biosens. Bioelectron., 2013, 41, 736 - 744.
322. K. Y. Inoue, M. Matsudaira, M. Nakano, K. Ino, C.
Sakamoto, Y. Kanno, R. Kubo, R. Kunikata, A. Kira, A. Suda,
R. Tsurumi, T. Shioya, S. Yoshida, M. Muroyama, T.
Ishikawa, H. Shiku, S. Satoh, M. Esashi and T. Matsue, Lab
Chip, 2015, 15, 848 - 856.
323. H. M. Jafari, K. Abdelhalim, L. Soleymani, E. H. Sargent, S.
O. Kelley and R. Genov, IEEE J. Solid-State Circuit, 2014,
49, 1223 - 1241.
324. A. Ganguli, Y. Watanabe, M. T. Hwang, J.-C. Huang and R.
Bashir, Biomed. Microdevices, 2018, 20, 45.
325. J. M. Rothberg, W. Hinz, T. M. Rearick, J. Schultz, W.
Mileski, M. Davey, J. H. Leamon, K. Johnson, M. J.
Milgrew, M. Edwards, J. Hoon, J. F. Simons, D. Marran, J.
W. Myers, J. F. Davidson, A. Branting, J. R. Nobile, B. P.
Puc, D. Light, T. A. Clark, M. Huber, J. T. Branciforte, I. B.
Stoner, S. E. Cawley, M. Lyons, Y. T. Fu, N. Homer, M.
Sedova, X. Miao, B. Reed, J. Sabina, E. Feierstein, M.
Schorn, M. Alanjary, E. Dimalanta, D. Dressman, R.
Kasinskas, T. Sokolsky, J. A. Fidanza, E. Namsaraev, K. J.
McKernan, A. Williams, G. T. Roth and J. Bustillo, Nature,
2011, 475, 348 - 352.
326. C. Toumazou, L. M. Shepherd, S. C. Reed, G. I. Chen, A.
Patel, D. M. Garner, C.-J. A. Wang, C.-P. Ou, K. Amin-Desai,
P. Athanasiou, H. Bai, I. M. Q. Brizido, B. Caldwell, D.
Coomber-Alford, P. Georgiou, K. S. Jordan, J. C. Joyce, M.
L. Mura, D. Morley, S. Sathyavruthan, S. Temelso, R. E.
Thomas and L. Zhang, Nat. Methods, 2013, 10, 641 - 646.
327. T. Hayasaka, S. Yoshida, K. Y. Inoue, M. Nakano, T.
Matsue, M. Esashi and S. Tanaka, J. Microelectromech.
Syst., 2015, 24, 958 - 967.
328. J. Rothe, O. Frey, A. Stettler, Y. Chen and A. Hierlemann,
Anal. Chem., 2014, 86, 6425 - 6432.
329. D. L. Bellin, H. Sakhtah, Y. Zhang, A. Price-Whelan, L. E. P.
Dietrich and K. L. Shepard, Nat. Commun., 2016, 7, 10535.
330. B. Nemeth, M. S. Piechocinski and D. R. S. Cumming, Sens.
Actuators B, 2012, 171 - 172, 747 - 752.
331. P. Shields, B. Nemeth, R. B. Green, M. O. Riehle and D. R.
S. Cumming, IEEE Sens. J., 2012, 12, 2744 - 2749.
J. Name., 2013, 00, 1-3 | 31
 Lab
Please do onadjust
not a Chipmargins Page 32 of 32

ARTICLE Journal Name

332. K. Sawada, H. Nakazawa, S. Takenaga, T. Hizawa, M. View Article Online

Futagawa, F. Dasai, T. Sakurai, K. Okumura, T. Hattori and DOI: 10.1039/C9LC01112A
M. Ishida, IEICE Trans. Fundamentals, 2014, E97-A, 726 -
733.
333. M. Mujeeb-U-Rahman, M. H. Nazari and M. Sencan,
Biosens. Bioelectron., 2019, 124 - 125, 66 - 74.
334. H. Jiang, X. Zhou, S. Kulkarni, M. Uranian, R. Seenivasan
and D. A. Hall, San Diego, USA, 2018.
335. Y. Huang and A. J. Mason, Lab Chip, 2013, 13, 3929 - 3934.
336. W. Deng, Y. Dou, P. Song, H. Xu, A. Aldalbahi, N. Chen, N.
N. El-Sayed, J. Gao, J. Lu, S. Song and X. Zuo, J. Electroanal.
Chem., 2016, 777, 117 - 122.
337. A. Ainla, M. P. S. Mousavi, M.-N. Tsaloglou, J. Redston, J.
G. Bell, M. T. Fernández-Abedul and G. M. Whitesides,

Lab on a Chip Accepted Manuscript

Anal. Chem., 2018, 90, 6240 - 6246.
338. J. Aymerich, A. Márquez, L. Terés, X. Muñoz-Berbel, C.
Jiménez, C. Domínguez, F. Serra-Graells and M. Dei,
Biosens. Bioelectron., 2018, 117, 736 - 742.
339. P. B. Lillehoj, M.-C. Huang, N. Truong and C.-M. Hod, Lab
Chip, 2013, 13, 2950 - 2955.
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