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Skin-interfaced microfluidic devices with one-

Cite this: DOI: 10.1039/d0lc00400f
opening chambers and hydrophobic valves for
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sweat collection and analysis†

Yingxue Zhang,‡ab Yao Chen,‡a Jielong Huang,a Yangchengyi Liu,a Jinfeng Peng,c
Shangda Chen,a Kui Song,d Xiaoping Ouyang,a
Huanyu Cheng *b and Xiufeng Wang *ae

Received 17th April 2020,
Accepted 1st June 2020
DOI: 10.1039/d0lc00400f
rsc.li/loc
Soft, skin-interfaced microfluidic platforms are capable of capturing, storing, and assessing sweat chemistry
and total sweat loss, which provides essential insight into human physiological health. However, sweat loss
from the outlet of the microfluidic devices often leads to deviation of the measured concentration of the
biomarker or electrolyte from the actual value. Here, we introduce hydrophobic valves at the junction of
the chamber and the microfluidic channel as a new chamber design to reduce sweat evaporation. Because
the advancing front of the liquid in the hydrophilic microchannel is blocked by the hydrophobic valve, the
fluid flows into the chambers, forms the initial meniscus, and completely fills the chambers along the initial
meniscus. Fluid dynamic modeling and numerical simulations provide critical insights into the sweat
sampling mechanism into the chambers. With significantly reduced evaporation and contamination, the
sweat sample can be easily stored for a long time for later analysis when in situ analysis is limited.
Additionally, the design with multiple chambers can allow sequential generation of sweat collection at
different times for long-term analysis. The in situ real-time measurements of the sweat loss and pH value
analysis from the human subject demonstrate the practical utility of the devices in collecting, storing, and
analyzing the sweat generated from sweat glands on the skin.

1. Introduction
Recent advances in materials, mechanics design, and device
architectures have spurred the rapid development of devices
that are thin, soft, and mechanically compliant. Capable of
interfacing with the human epidermis, this class of emerging
electronics offers continuous recording of vital physiological
parameters with clinical quality for health and performance
assessment. Sweat, which can be measured noninvasively and
contains various biomarkers, is starting to gain popularity for
physiological health monitoring and disease diagnostics.1–5
a
School of Materials Science and Engineering, Xiangtan University, Xiangtan,
Hunan 411105, China. E-mail: onexf@xtu.edu.cn
b
Department of Engineering Science and Mechanics, The Pennsylvania State
University, University Park, PA 16802, USA. E-mail: huanyu.Cheng@psu.edu
c
School of Mechanical Engineering, Xiangtan University, Xiangtan, Hunan 411105,
China
d
College of Civil Engineering and Mechanics, Xiangtan University, Xiangtan,
Hunan 411105, China
e
Institute of Flexible Electronics Technology of Tsinghua University, Jiaxing,
Zhejiang 314006, China
† Electronic supplementary information (ESI) available. See DOI: 10.1039/
d0lc00400f
‡ Y. Z. and Y. C. contributed equally to this work.
For instance, local and full-body sweat loss rates provide
essential insights into the hydration state of an individual.6,7
Quantitative measurements of the sweat rate can prevent
heat-related illnesses, including heat stroke and heat
exhaustion, in hot environments.8 Because the pH of sweat is
associated with metabolic alkalosis, noninvasive
measurement of metabolic alkalosis from perspiration is
possible.9,10 Additionally, sweat electrolytes and metabolites
(e.g., lactate, chloride, and glucose) can be used to inform
electrolyte balance, blood glucose levels,9,11 hydration status,
and overall health.12
Early developed sweat collection devices rely on absorbent
pads made from filter paper, cotton, gauze, or toweling.13
After separating the sweat from the absorbent pads by
centrifugation, sweat analysis is performed with
corresponding benchtop equipment.14 However, this process
requires expensive equipment and trained professionals to
collect and analyze the data.15 As an alternative, wearable
microfluidic devices open opportunities to collect and analyze
sweat in situ for personalized medicine, improved sports
performance, and military readiness.16–21 For commonly used
microfluidic channels that are hydrophobic, the sweat must
overcome the positive capillary pressure to enter the
microchannels for capture and collection. In contrast, the

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hydrophilic microfluidic channels that are associated with
negative capillary pressure spontaneously wick sweat into the
microchannels more rapidly by capillary action.
Due to the inherently low concentrations of biomarkers
and electrolytes (e.g., sodium, chloride, and potassium), their
measurements are susceptible to evaporation.22 Silicone
elastomers such as poly(dimethylsiloxane) (PDMS) are
commonly explored as microfluidic devices with an ultrathin
capping layer of 150 μm for sweat sampling; they are
associated with ∼100% water loss within 3 hours at 37 °C.12
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However, PDMS is still commonly used as a microfluidic
material due to its biocompatibility, optical transparency,
ease of patterning, and favorable mechanics. Depending on
the design of the microfluidic device, the measured
concentration of the biomarker or electrolyte can either
slightly or greatly deviate from the actual value because of
water loss. Most of the previous designs of microfluidic sweat
sampling devices exploit chambers with valves2,3,22 as well as
one inlet for sweat collection and one outlet connected to the
surroundings for releasing the air in the chambers. While the
outlet avoids backpressure, it also leads to sweat evaporation
because most evaporation in the chambers occurs through
the outlet.23 Thus, a novel design of a microfluidic sweat
sampling device to reduce unnecessary evaporation and
contamination of sweat in the collection chambers is vital for
accurate sweat analysis.
As one of the most critical components in microfluidic
devices, valves are commonly used to control the direction of
the flow.24 Traditional valves with external device
components to drive liquid flow are bulky and thus
challenging for direct integration on the skin.25 In contrast,
passive valves without active components or power sources
allow for fluid control, presenting an attractive option for
wearable devices. Representative passive valves include
capillary bursting valves,26 super-absorbent polymer valves,22
and hydrophobic valves22 to guide the flow in a well-defined
sequence. Capillary bursting valves are among the most
common passive valves in wearable devices to control the
sweat flow by reducing the width of the microfluidic
channels, which are associated with higher bursting pressure.
The preparation of the capillary bursting valves often involves
lithographic processes, which complicates the fabrication
and substantially increases the cost in rapid prototyping.27
Based on super-absorbent polymer (SAP) materials prepared
by a cross-linking reaction, the SAP valve swells upon
hydration to close the inlet and prevent further flow.
However, the preparation of the SAP material is complex, and
additional steps are also needed for integration of the device.
By contrast, hydrophobic valves can be readily fabricated in a
high-density array. The hydrophobic valves explored in
previous literature reports22,28 were only used to guide the
flow in microfluidic channels. In contrast, we report the
design concepts, fluid mechanics principles, and applications
of hydrophobic valves in microfluidic devices with one-
opening chambers which can simply achieve the chrono-
sampling of liquid/sweat. The hydrophobic valves reported by
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L. Riegger et al.29 were generated by enhancing the
hydrophobicity of the PDMS surface, which is associated with
complex fabrication processes and a much higher bursting
pressure (2.5–4.5 kPa). Because this bursting pressure is
higher than the sweat pressure, hydrophobic valves cannot
be easily integrated into microfluidic devices for sweat
collection. In contrast, endowing the rest of the microfluidic
device with hydrophilicity allows us to directly explore the
use of naturally hydrophobic PDMS in hydrophobic valves.
The fabrication process is simple and effective to generate
hydrophobic valves with long-term stability. Additionally, the
hydrophilic microfluidic devices in other locations can also
spontaneously wick sweat into the microfluidic channels.
Here, we introduce hydrophobic valves at the junction of
the chamber and the microfluidic channel as a new chamber
design to reduce sweat evaporation. The hydrophobic valves
prepared by air plasma treatment guide the sweat flow to fill
the chambers sequentially. Because the advancing front of
the liquid in the hydrophilic microchannel is blocked by the
hydrophobic valve, the fluid flows into the chambers, forms
the initial meniscus, and completely fills the chambers along
the initial meniscus. Fluid dynamic modeling and numerical
simulations provide critical insights into the mechanism of
sweat sampling into the chambers. With significantly
reduced evaporation and contamination, the sweat sample
can be easily stored for a long time for later analysis when in
situ analysis is limited. Additionally, the design with multiple
chambers can allow sequential sweat collection at different
times for long-term analysis.
2. Results and discussion
2.1. Soft microfluidic device with hydrophobic valves for
sweat collection
Fig. 1a presents an exploded view of the novel microfluidic
device designed for accurate sweat collection and analysis
along with a schematic of its working principle. The
representative device is 0.84 mm thick and is designed in a
semicircular geometry with a diameter of 2.3 cm to minimize
the overall footprint. The proof-of-concept device includes
three chambers for the analysis of the chloride concentration,
pH, and glucose concentration because these are common
biomarkers in sweat. Depending on need, the number of
chambers can be easily modulated for the target application.
Following the fabrication process (Fig. S1†) yielded
microfluidic devices that consisted of three layers: 1) a
capping layer, 2) a microfluidic layer, and 3) a medical-grade
acrylic adhesive film for enhanced adhesion to the skin.
Because PDMS with a low effective modulus of ∼145 kPa is
biocompatible and optically transparent, it was explored for
both the capping and microfluidic layers. The capping layer
with a thickness of 240 μm avoided self-collapse of the
microfluidic channel (Fig. S2†). The microfluidic layer with a
thickness of 600 μm included a microfluidic channel (300
μm in width and height, consistent with previous designs2)
and chambers with a diameter of 3 mm. The surface of the
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capillary pressure in the hydrophilic microfluidic channels.

The device with a small thickness, soft mechanics, and a
tapered edge avoids delamination from the skin12 and
damage upon mechanical deformations such as bending and
twisting (Fig. 1b). Additionally, the device is positioned on
the skin with little deformation or strain (e.g., forehead,
forearm, and back). In addition to robust adhesion, the
adhesive layer also helps eliminate leakage during natural
skin motions from daily activities or vigorous physical
exercise.
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2.2. Hydrophobic valves in hydrophilic microfluidic channels

Fig. 1 Schematic and optical images of an epidermal microfluidic
device with one-opening chambers and hydrophobic valves (HVs) for
chrono-sampling of liquid. (a) Schematic of an exploded view of the
device, highlighting various functional layers and working principles of
the HVs. (b) Optical images of the microfluidic device on the skin and
under mechanical deformations such as bending and twisting.
While sweat excretion generates positive pressure, it must
still overcome the capillary pressure to enter the hydrophobic
microfluidic channels. In this study, we designed and
demonstrated hydrophilic microfluidic channels with
negative capillary pressure to spontaneously wick sweat into
the channel. Following the fabrication of a single
microchannel (Fig. S1†), its surface wettability was first
investigated. The silicone polymer PDMS was naturally
hydrophobic, with an intrinsic water contact angle (CA) of
107° (Fig. 2a, red line). After exposing the surface to air
plasma treatment (APT), the PDMS microfluidic channel

microfluidic channel was measured to have low roughness,

and the arithmetic mean height (Sa) was calculated to be
0.43 μm (Fig. S3†), comparable to that from literature reports
of 0.2 μm.30 Compared to the height of the microfluidic
channel (i.e., 300 μm), the much smaller surface roughness
exhibited a negligible effect on the sweat collection.
Additionally, the surface roughness of the PMMA mold did
not affect the PDMS bonding (Fig. S1†) because of the
demonstrated robust device performance during use even
after intensive exercise. By blocking the sweat flow, the
hydrophobic valves help route sweat into the chambers and
then open to allow sweat flow after the chambers are
completely filled. Because the chambers are filled
sequentially (labeled T1, T2, and T3 based on their filling time
points), the microfluidic device provides effective chrono-
sampling capability for in situ analysis of various biomarkers
and electrolytes. The inlet hole with a diameter of 3 mm
(corresponding to ∼9 sweat glands26) in the watertight
adhesive layer defines the localized sweat collection regions
from the skin sweat glands. The geometry of the microfluidic
channel and chambers in the device determines the sweat
Fig. 2 Design of hydrophobic valves in hydrophilic microfluidic
collection volume. After considering the target application channels. (a) The water contact angle of the PDMS surface as a
with a prescribed monitoring time, the number of sweat function of aging time before and after various surface treatments. (b)
glands within the device inlet can then be determined. The (i) Schematic of the liquid blocked by a functional hydrophobic valve in
proof-of-concept demonstration in the current study selects 9 a hydrophilic microfluidic channel. Photographs showing the liquid in
hydrophilic microfluidic channels prepared by (ii) plasma-treated PDMS
sweat glands for the chosen device geometry and a target
or (v) plasma and PVP-treated PDMS and blocked by the HV inside the
monitoring of 1 h. The sweat is driven to flow through the channel, along with (iii) the droplet residing on the inlet hole of an
inlet hole into the microfluidic system by the positive untreated hydrophilic channel and (iv) the liquid passing over the
pressure from natural sweat excretion and the negative hydrophilic channel without a hydrophobic valve.

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becomed hydrophilic and generated negative capillary
pressure, which quickly wicked the liquid into the channel
(Fig. 2b-ii). In contrast, the hydrophobic microfluidic channel
could not imbibe the droplet to wick it into the channel
(Fig. 2b-iii and S4†). However, the intrinsic hydrophobicity of
PDMS gradually recovered ca. 5 h after the APT (Fig. 2a,
purple line). To address this challenge, subsequent low-energy
polyvinylpyrrolidone (PVP) surface modification of the APT-
treated PDMS was exploited. While the contact angle still
increased from less than 20° to ca. 42° ± 2°, it saturated at this
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value and remained stable for more than 24 h (Fig. 2a,
blue line). The hydrophilic surface of the PDMS after the
oxygen plasma treatment and PVP modification can remain
stable for 6 months.31 Therefore, the shelf life of the
microfluidic device is over 6 months. This long-term stability of
the hydrophilic surface is attributed to the presence of CO as
evidenced in the FTIR spectra (Fig. S5†) from the PVP
treatment, whereas the APT-treated PDMS only temporarily
changes the density of the –OH groups (water-adsorbed) at its
surface. This hydrophilic microfluidic channel from the APT
and PVP modification easily induced imbibition of fluid into
the channel (Fig. 2b-iv).
Modulation of the sweat flow in the hydrophilic
microfluidic channel was achieved by hydrophobic valves
(HVs), which were simply created using a mask to avoid
exposure of PDMS to air plasma during the APT (Fig. S1†).
The unexposed PDMS region under the mask remained
hydrophobic and formed the HV in the microfluidic channel.
Because the PVP surface modification in the microfluidic
channel was selective only for the hydrophilic region, the
obtained HV was not affected by the PVP surface
modification. In contrast, rendering the rest of the
microfluidic device hydrophilic allowed us to directly explore
the use of the naturally hydrophobic PDMS in hydrophobic
valves. The fabrication process was simple and effective to
generate hydrophobic valves with long-term stability.
Additionally, the hydrophilic microfluidic device in other
locations could also spontaneously wick sweat into the
microfluidic channels. In the proof-of-concept
demonstration, an HV with a length of l = 1 mm and a width
and height of w = h = 300 μm was patterned at a specific
distance (i.e., 2 cm) away from the inlet hole (Fig. 2b-i). The
fluidic flow in the hydrophilic microfluidic channel was
successfully stopped by the HV, as the air–liquid interface in
the channel was observed under a microscope (Fig. S6†). The
impeditive function (PHV) of the HV can be explained by the
capillary theory (Fig. S7†):28,32
 1
πð1 − cos θ0 Þ2 ð4 − cos θ0 − 3 cos2 θ0 Þ 3
P HV ¼ 2σ
 6V  geometry changes and reduce the fluid resistance. The
w 2h þ w
þσ cos θ1 þ cos θA ≤ 0 (1)
wh wh
where V is the droplet volume, σ is the surface tension of the
liquid, and θ0 and θA are the contact angles (CAs) experienced
by the fluid at the top inlet and the HV, respectively. The first
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term in eqn (1) is contributed by the droplet volume at the
inlet, whereas the second term is related to the hydrophilic
top wall (θ1 = 42°) and three other hydrophobic walls (i.e., the
1st and 2nd terms in the parenthesis, respectively). The CA at
the HV is the intrinsic contact angle of PDMS (i.e., θA = 107°),
whereas the CA at the top inlet depends on the treatment of
the PDMS microfluidic channel (i.e., θ0 = 107°, 90°, or 42° for
the untreated, APT-treated, and APT-treated with PVP-
modified PDMS, respectively). The impeditive function on the
left-hand side of eqn (1) decreases as the contact angle
experienced by the HV increases (Fig. S7†). Because the
bursting pressure of the HV is simply the second term in eqn
(1), the bursting pressure of the HVs with a contact angle of
107° is calculated to be 0.04 kPa (Fig. S8,† yellow star). When
the top wall is also rendered hydrophobic, the bursting
pressure will increase from 0.04 kPa to 0.28 kPa, representing
a simple way of tuning. As eqn (1) holds true for all three
types of microfluidic channels even for an extremely small
droplet volume (V = 0.5 μL), the HV works effectively as
designed (Fig. 2b-ii and v), whereas the fluid flows through
the entire hydrophilic channel without the HV (Fig. 2b-iv).
2.3. Working principle of HVs and their use in sequential
liquid sampling
Because chambers in microfluidic devices are often exploited
for sweat collection, novel one-opening microfluidic
chambers with an HV in the bridging channel were designed
(Fig. 3a). A unit of the microfluidic devices consists of the
main channel, a bridging channel, a collection chamber, and
an HV. In one embodiment, the bridging channel includes a
neck microchannel (length of 80 μm and width of 500 μm)
and two transition channels with radii of 300 μm (Fig. S9†).
The longer neck microchannel forms a narrower gas outlet
between the liquid meniscus and the bridging channel,
which traps air to increase the backpressure in the chamber.
For instance, the trapped air in the chamber prevented liquid
collection in the chamber when the length of the neck
microchannel was increased from 80 μm in the current
design to 200 μm (Fig. S9c†). On the other hand, a significant
diffusion between two different fluids occurred as the width
of the neck microchannel increased from 0.5 mm in the
current design to 2 mm (Fig. S9d†). Because the preparation
of a shorter and narrower neck microchannel was associated
with higher fabrication complexity and cost, the neck
channel with a width of 0.5 mm and a length of 80 μm was
chosen in this study. After the geometries of the neck
channel and chamber were chosen, the transition channels
between the two were then determined to avoid drastic
design of the HV is also important to allow the release of air
from the chambers to avoid backpressure. Although the
geometry of the HV is determined by the microfluidic
channel (Fig. S7†), its position in the main microfluidic
channel also affects liquid collection in the chambers. The
HV (see dimensions in Fig. S9†) located in the microchannel
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Fig. 3 Working principle of the hydrophobic valve to route liquid into the one-opening chamber in the microfluidic device. (a) Schematic of the
one-opening chamber with the hydrophobic valve and a bridging channel in a microfluidic device. (b) Optical images (i–iv) and (c) numerical
simulations of the hydrodynamic flow process into the one-opening chamber with a hydrophobic valve, compared to (d) optical images (i–iv) and
(e) numerical simulations of the one-opening chamber without a hydrophobic valve.

needs to be positioned close to the chamber according to the
analysis in the following section.
The capability of the novel one-opening microfluidic
chambers with the HV in the bridging channel was
demonstrated through a series of optical images during a
fluid collection process (Fig. 3b). Applying a flow rate of 1.0
μL min−1 as in the previous in vitro experiments,23 red-dyed
water was pumped into the hydrophilic microfluidic channel.
The relatively low flow rate enabled easy observation of the
fluid collection. Because the HV remained closed before the
pressure across it exceeded the threshold,33 it routed the
fluid to flow into the chamber (Fig. 3b). Observation of the
air–liquid interface (i.e., the two-point contact labeled A and
B) in the microfluidic channel illustrated the fluid collection
process. As the liquid initially arrived at the HV, it was
blocked; the air–liquid interface at point B was pinned at the
HV, whereas the air–liquid interface at point A was forced
through the bridging channel into the chamber. As point A
moved along the chamber wall clockwise and was pinned at
the HV, an initial liquid meniscus formed on the chamber
wall (Fig. 3b-ii). Due to the pinning of the air–liquid interface
at both points A and B, the liquid meniscus continued to
increase (Fig. 3b-iii) and filled the entire chamber (Fig. 3b-iv).
After the liquid filled the bridging channel, the air–liquid
interface with sufficient pressure burst open the HV and
continued to flow.
As a comparison, the one-opening microfluidic chambers
without the HV failed to collect the sweat in the chamber, as
demonstrated in a series of optical images during the fluid
collection process (Fig. 3d). The air–liquid interface first
moved through the bridging channel and formed a sloped
liquid meniscus (Fig. 3d-ii). Continuing of the fluid flow split
the air–liquid interface into two meniscuses: 1) a horizontal
meniscus that enclosed the chamber and trapped the air
inside, and 2) a vertical meniscus that continued to move in
the main channel. Due to insufficient pressure to compress
the air in the chamber, the horizontal liquid meniscus failed
to fill the chamber (Fig. 3d-iv). The designed one-opening
microfluidic chambers with and without the HV also showed
reasonably good agreement with the 2D numerical

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Fig. 4 Functional demonstrations of the microfluidic devices (a) with black-dyed water and (b) with two colors of water dyed in red and blue (the
zoomed-in image of the 3rd chamber shown in the inset indicates minimal mixing). (c) Measurements of the water loss in the chambers with and
without an outlet, indicating reduced evaporation for improved accuracy of analyte assessment.
simulations (Fig. 3c and e), which further verified the
feasibility of the proposed designs.
Next, sequential sweat collection was demonstrated for
potential applications in which it was necessary to analyze
the concentration changes of various biomarkers and
electrolytes at different time points (Fig. 4). The chrono-
sampling process was illustrated by spontaneous and
sequential pumping of black-dyed water into the hydrophilic
microfluidic channel (Fig. 4a and Movie S1†). The flow rate
was found to be 60 μL min−1 as the ratio of the volume in the
microfluidic channel (0.5 μL, blue region in the following
figure) to the corresponding filling time of 0.5 s. Although
the function of the HVs may be compromised when operated
at an even higher flow rate, the maximum sweat rate in the
inlet (5.4 μL min−1 generated by 9 sweat glands, each with
the highest rate of 0.6 μL min−1 per gland as in previous
reports23) is well within the demonstrated operation rage.
Because the top wall in the HV is hydrophilic in the current
demonstration, a small amount of the black-dyed water
wicked into the main microfluidic channel before the
chamber was completely filled. However, this has a negligibly
small effect on the sweat collection. Endowing the top wall of
the HV with hydrophobicity could also eliminate or minimize
this effect. Minimal interference with the initially collected
sweat sample from the later collection was also demonstrated
using water dyed with two colors (Fig. 4b). In this
demonstration, the 1st chamber was filled with red-dyed
water, whereas the 2nd and 3rd chambers were subsequently
filled with blue-dyed water. The diffusion between the two
different fluids occurred in two directions: 1) from the 2nd
fluid in the main microfluidic channel to the 1st fluid in the
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chamber, and 2) from the 1st fluid in the chamber to the 2nd
fluid in the main microfluidic channel. As demonstrated in
Fig. 4b, the diffusion along these two directions occurred only
at the chamber wall, with less than 5% of the total volume in
both the 1st and 2nd chambers (Fig. S10†). Because the
volume of the red-dyed water was larger than that of the 1st
chamber, it was partially collected by the 2nd chamber, with a
slightly larger mixing of 15%. The design of the 2nd chamber
was indeed chosen to buffer the transition between the two
different fluids. Because of the micro-scale dimensions in the
device and low sweat flow rates, molecular diffusion
accounted for the mixing inside the chambers.23 The
diffusion (or interaction) between the two different fluids only
occurred at the chamber wall (less than 5% of the total
volume of a collection chamber), indicating minimal
interference between fluid collections at different time points.
This feature also allowed accurate assessment of multiple
biomarkers in different chambers with a single device.
The novel one-opening microfluidic chambers also
reduced the evaporation rate of the fluid inside the chamber.
The weight loss of the liquid was determined by the
difference between the remaining weight of liquid V and the
initial weight of liquid V0; both were measured by a high-
precision electronic balance. Although weight loss was
observed in both chambers when they were exposed to air at
26 °C, the one-opening chamber (i.e., without an outlet)
exhibited remarkably smaller loss than that with an outlet
(0.45 mm) over a period of 10 h (Fig. 4c). Therefore, the
outflow evaporation rate was much smaller than the inflow
sweat rate, allowing the sweat to fill the chamber during the
low-intensity exercise. The storage time is already sufficient
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for many applications (e.g., in situ colorimetric analysis or

short-term storage before ex situ analysis). Additionally, the
storage time can be significantly increased by reducing the
storage temperature or covering the outlet. It should be noted
that the sweat collection would be challenging if the sweat rate
were significantly reduced. Although our new design of a one-
opening chamber could help reduce the evaporation rate,
efforts are still needed to address the challenges in this new
application. Because sweat evaporation inevitably increased the
measured concentrations of the biomarkers or electrolytes (10–
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100 mM for sodium in sweat1), the one-opening chamber led

to a concentration measurement ca. 1.6 times more accurate
than those with an outlet after 3 h. To demonstrate the
increased sensing accuracy from the reduced evaporation rate,
a pH assay (902 07, Macherey-Nagel, Germany) was used as
evaporation from the chambers occurred at 40 °C (Fig. S11†). A
large deviation in the R index of ∼10 was first observed
between the two designs (i.e., with and without the 0.45 mm
outlet). Because the pH value was calculated from the RGB
index, the pH value at the measured time point would be
different from the time of collection due to evaporation. Fig. 5 Fluid dynamic modeling of the fluid collection process.
Therefore, a reduced evaporation rate would result in a better Schematics of the advancing front of the air–fluid interface into the
chamber (a) with and (c) without the hydrophobic valve. (b) Calculated
representation of the pH value at the time of collection.
map of the normalized capillary driving pressure versus the normalized
location variables of the advancing interface (l′ and x′). The yellow line
2.4. Fluid dynamic modeling of one-opening chambers with represents the calculated motion trajectory of the advancing front of
the HV the air–fluid interface based on the fluid dynamic model. The points
mark the experimental and simulated results, with the circle or cross
The pressure across the air–liquid interface in the channel corresponding to the success or non-success filling of the chamber.
(Pi) is obtained as (see Methods in the ESI†):
 
sinðα − θA Þ cos θA
P i ¼ 2σ þ (2)
b* h Pb ðL′ þ l′ − 0:5πÞ cos θA þ ð1 − x′Þ sin θA
r¼ 2
; (3b)
2σ L′ þ l′2 þ 0:25π2 − L′ðπ − 2l′Þ − πl′ þ ð1 − x′Þ2
where σ is the surface tension of the liquid, θA is the constant π π 1
≤ l′ ≤ þ ;
contact angle at the channel wall, α is the apparent angle 2 2 4
between the tangent to the channel wall and the straight line
of the air–liquid interface, and b* and h are the effective where L′ is defined as L′ = L/r and x′ is related to l′ through eqn
width and height of the channel, respectively. The capillary (S5) in the ESI† (Fig. S12†). The normalized driving pressure Pb
pressure difference (ΔP) that drives the liquid advancing front r/2σ is positive for the APT-treated and PVP-modified

Pb ð4 þ L′Þ cos θA − cosð0:25 − l′ − θA Þ − x′ sin θA π 1 1

r¼ 2 2
; þ ≤ l′ ≤ π þ : (3c)
2σ ð4 þ L′Þ þ ð1 þ x′ Þ − ð8 þ 2L′Þ cosð0:25 − l′Þ þ 2x′ sinð0:25 − l′Þ 2 4 4

into the chamber is mainly contributed by Pi in eqn (2). As the
air–liquid interface advances in the bridging channel, the
effective channel width increases and the apparent contact
angle α decreases in eqn (2). As a result, the first term in eqn
(2) changes as the air–liquid interface advances in the bridging
channel, whereas the second term is independent of the
position. Therefore, the first term of the driving pressure (i.e.,
Pb = 2σ sin(α − θA)/b*) has then been systematically investigated.
Denoting the advancing front of the air–liquid interface at l′ = l/
r and x′ = x/r with r as the radius of transition channels
(Fig. 5a), the normalized driving pressure is obtained as
Pb L′ cos θA − cosðl′ þ θA Þ − x′ sin θA π
r¼ ; 0 ≤ l′ ≤ ; (3a)
2σ 1 þ L′2 þ x′2 − 2L′ cos l′ − 2x′ sin l′ 2
hydrophilic microfluidic channel (θA = 42° ± 2° in Fig. 5b)
during the process of the liquid flow into the chamber.
Considering the additional constant driving pressure, the
resulting driving pressure spontaneously wicks the fluid into
the one-opening chamber with the HV.
In the case of the one-opening chamber without the HV,
the resistance of compressed gas is estimated to be ca. 6 kPa
upon filling of the bridging channel (Fig. 5c). Because the
resistance is one order of magnitude higher than the
capillary drive force, the liquid does not enter the chamber.
In contrast, placing the HV near the chamber routes the fluid
into the chamber. Three experimental measurements and five
calculation cases (see details in Fig. 3 and S13†) marked in
Fig. 5b indicate the critical position of the HV at x′c = 3.5 (i.e.,

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the HV needs to be placed to the left of x′c ). The HV placed
to the left of the critical position allows the sweat to flow
along on the chamber wall, which can effectively vent air
through the HV to avoid backpressure. When the HV is
placed to the right of the critical position, the chamber is
not filled and the liquid simply flows through the main
microfluidic channel. Once the chamber is filled with the
liquid, the flow automatically bursts open the HV. During
the liquid collection at a later stage, the liquid bypasses the
previous chamber without mixing with the liquid samples
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2.5. Chrono-sampling and chemical analysis of the sweat
Human testing involved the evaluation of the skin-interfaced
microfluidic device mounted on the forehead of a young
adult volunteer during running exercises (Fig. 6a). The sweat
was driven into the microfluidic device by the positive
pressure from sweat excretion, assisted by the capillary action
in the hydrophilic channels. The flow rate of the sweat in the
microfluidic channel was determined by the dimension of
the microchannel, surface tension, and the positive pressure

collected in the previous chambers. Due to its large
curvature, the curved main microfluidic channel is
approximated to a straight channel for easy calculation. The
simulated results further indicate no significant differences
for sweat collection between the circular and straight main
microfluidic channel (Fig. S13†). Because of the curved
main microfluidic channel, the effective width experienced
by the liquid meniscus could be slightly different from the
theoretical model. According to a previous literature
report,34 the calculated bursting pressure would differ by
<5%. Because the calculated bursting pressure is much
smaller than the pressure generated by the eccrine sweat
gland (i.e., 2.4–2.9 kPa), the effect from this small variation
is minimal.
from sweat excretion. The sweat flow rate from the young
adult volunteer may not be constant; however, a relatively
stable flow advancing front during the low-intensity running
exercise was observed, indicating a relatively small effect
from the varying sweat flow rate. It was also noted that the
largely varied flow rate may lead to different timeframes to
completely fill each chamber; however, the time point for
analysis could be adjusted accordingly. Although running
could cause vibration, the relatively strong adhesion strength
between the device and skin confined the sweat within the
microfluidic device. Placing a red dye of water-soluble
particles at the inlet of the microfluidic device allowed for
visual assessment of the cumulative sweat loss during the
sweat collection process. The red-dyed sweat was observed to

Fig. 6 In situ chrono-sampling from various body positions during running exercise. (a) Images of microfluidic devices applied to the forehead for
sweat collection and (b) the corresponding volume measurements through a syringe pump study. Note: the syringe pump study does not include
the volume of ∼6 μL in the inlet; therefore, it must be added before filling of the 1st chamber in the human testing. (c) Changes in the RGB values
as a function of pH value. (d) Images of microfluidic devices deployed to the lower back over half an hour. (e) The sequentially measured sweat pH
values from various body positions (i.e., lower back, neck, and forehead).

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Lab on a Chip
be confined within the microfluidic channel and chambers
even during low-intensity physical exercise for ca. one hour,
consistent with literature reports.2,9,23 The robust adhesion
strengthened from the chosen adhesive layer avoided the
detachment of the device from the skin. However, long-term
use of the device for sweat monitoring may result in the
detachment, which will be systematically studied in the
future. With a volume capacity of 2.12 μL in each chamber
and a capacity of 3.64 μL in the microfluidic channel, the
total volume of ∼10 μL (verified by a syringe pump study in
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Fig. 6b) in the microfluidic device corresponded to the
quantitative operational requirements. By taking the ratio of
the volumes (Fig. 6b) to times (Fig. 6a) after the sweat filled
each chamber, the average sweat rate from each sweat gland
(9 in total) was calculated to be 0.065, 0.04, and 0.018 μL
min−1 from the three chambers during the chrono-sampling
over the running exercise. Notably, the peak sweat rate from
the measurement of the 1st chamber corresponded to the
initial stage of the exercise with the largest sweat rate. The
average sweat rate per sweat gland during the entire period
of the exercise was estimated to be 0.04 μL min−1. The
measured effective sweat rate is larger than the value
measured by conventional absorbent pads (0.006 μL min−1),35
mainly due to compensatory effects (i.e., higher sweat rates
in the open sweat glands because the adjacent regions are
blocked by the adhesive).23 Nevertheless, the increased
effective sweat rate helps increase the speed of the sweat
collection in the microfluidic device.
While the collected sweat samples in these chambers can
be retrieved and analyzed later when in situ analysis is
limited, instantaneous and quantitative chemical assessment
of the sweat is also possible through colorimetric schemes.
As a simple yet representative demonstration, the pH of
sweat in the different chambers was analyzed using methyl
red as a pH indicator. Calibration of paper-based colorimetric
pH assays with buffer solutions over a medically relevant
range (pH from 4.5 to 6.0) indicated a significant change in
the G value among the three RGB values (Fig. 6c). Mounting
the microfluidic devices on various body positions (i.e., lower
back, neck, and forehead) quantified the sweat pH over half
an hour (lower-back, Fig. 6d). Using the calibration curve
(Fig. 6c), the G value obtained by the software from the
digital color images was converted into concentrations of the
target analyte (e.g., pH). The sequentially measured sweat pH
was also different among the lower back, neck, and forehead
(Fig. 6e). While the sweat pH from the lower back
sequentially decreased, no substantial differences were
observed in the sweat pH from the forehead and neck,
providing critical insights into the underlying physiological
variations.
3. Conclusions
This paper presents the design and demonstration of a novel
skin-interfaced microfluidic device with one-opening
chambers and hydrophobic valves for sweat collection and
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Paper
analysis. By reducing sweat evaporation, the new chamber
design with only one opening provides a more accurate
assessment of the biomarker or electrolyte in the sweat. Fluid
dynamic modeling and numerical simulations have been
explored to validate the effectiveness of the design. Human
testing on young healthy volunteers demonstrates the
chrono-sampling capabilities of sweat collection, storage, and
analysis. Taken together, the microfluidic device with one-
opening chambers and hydrophobic valves with significantly
reduced fabrication complexity provides a simple alternative
route to study sweat physiology for healthcare.
4. Materials and methods
4.1. Materials
Materials used in this study include polymethylmethacrylate
(PMMA; Microchem, MA, China), polydimethylsiloxane
(PDMS; Sylgard 184, Dow Corning, US), surface treating agent
(JD-206, Dongguan Jiudian Co. Ltd, China), adhesive film
(IV300, Smith & Nephew Medical Ltd, UK), polyvinyl
pyrrolidone (PVP, Sinopharm Chemical Reagent Co., Ltd,
China), methyl red (Tianjin Guangfu Fine Chemical Research
Institute, China), hydrogen phosphate (Sinopharm Chemical
Reagent Co., Ltd, China), and acid monohydrate (Aladdin
Industrial Corporation).
4.2. Design and fabrication of the microfluidic device
The microfluidic devices consisted of three layers: a PDMS
cover, a microfluidic layer, and an ultrathin adhesive layer.
The microfluidic microchannels and chambers on the surface
of polymethylmethacrylate (PMMA; Microchem, MA, China)
were created by micromachining (CNN, JK-DK40, Wenhao
Co. Ltd, China). Polydimethylsiloxane (PDMS; Sylgard 184,
Dow Corning, US) was mixed at a 20 : 1 ratio by weight to
prepare the PDMS cover and microfluidic layer. Pouring 0.5 g
of the mixture into the PMMA molds, followed by degassing
in a vacuum desiccator for removing bubbles and curing at
80 °C for 50 min, formed the microfluidic layer with a
thickness of ∼600 μm. The PDMS cover with a thickness of
∼300 μm was created by spinning (300 rpm for 30 s on a
smooth PMMA, curing at 80 °C for 50 min). The pH assay
papers were loaded into the chambers. The PDMS cover and
microfluidic layer were bonded by exposure to air plasma
(Harrick Plasma Cleaner, PDC-002-HP, Harrick Plasma, US)
with ‘high’ power for 3 min. Next, the surface of the device
was covered with a surface treating agent (JD-206, Dongguan
Jiudian Co. Ltd, China). A double-sided adhesive was applied
to connect the adhesive layer to the device. A thin adhesive
film (IV300, Smith & Nephew Medical Ltd, UK) was placed at
the bottom of the device to adhere to the skin. After the mask
for the HVs was designed by AutoCAD, a laser cutting system
(NARVEL 6000) was used to cut the mask from stainless steel.
Next, a microscope on the high-precision three-axis micron
positioning stage was used to place the mask at the
prescribed location in the main microfluidic channel close to
the chambers. Exposing the microfluidic device to air plasma
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Paper
treatment endowed the PDMS with hydrophilicity in the
regions not covered by the mask. The unexposed PDMS
region under the mask remained hydrophobic and formed
the HV in the microfluidic channel.
4.3. Surface modification
The PDMS layers were washed with ethanol and deionized (DI)
water, followed by drying with N2. The PDMS layers were
treated with air plasma with ‘high’ power for 3 min to activate
the surface. Next, the PDMS layers were immersed in 10% (w/v)
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polyvinyl pyrrolidone (PVP, Sinopharm Chemical Reagent Co.,
Ltd, China) solution for 3 min. After washing with DI water
and drying with N2, PDMS layers with long-term hydrophilic
surfaces were obtained. Spin casting of the 20 : 1 PDMS on a
smooth PMMA layer at 3000 rpm provided an uncured PDMS
thin layer. Dipping the capping layer against this uncured
PDMS thin layer generated a tacky surface on the capping layer
to facilitate its bonding with the microfluidic layer.
4.4. Contact angle measurements
The contact angles of the samples were measured using a
Model 250 (p/n 250-F1) optical contact-angle goniometer
(Rame-hart, USA) at ambient temperature. After dropping 3
μL of deionized water onto the sample surface with an
automatic dispense controller, the contact angles were
determined automatically using the Laplace–Young fitting
algorithm. The average contact angle was based on five
independent contact angle measurements.
4.5. Chemicals for the pH assay
Methyl red (Tianjin Guangfu Fine Chemical Research
Institute, China) served as a chromogen for pH detection. A
homogenous 0.5% (w/v) solution of methyl red was prepared
in ethanol. A filter paper served as the assay carrier of the
reaction and was hole-punched into circular shapes with
diameters of 2.5 mm. Next, each piece of filter paper was
dipped in the solution for 30 s and dried at room
temperature for 30 min to form a stable pH assay. Mixing 0.2
M disodium hydrogen phosphate (Sinopharm Chemical
Reagent Co. Ltd, China) and 0.1 M citric acid monohydrate
(Aladdin Industrial Co.) yielded pH buffer solutions with
various pH values (i.e., 4.5, 5.0, 5.5, and 6.0) as validated by a
pH meter (PHS-3B, Shanghai Shiheng Co. Ltd, China). After
dropping 1 μL of buffer solution on the pH essay papers,
Photoshop was used to determine the RGB values of each
chromogenic paper, which corresponded to
concentrations of the selected biomarkers.
4.6. Chrono-sampling of sweat in microfluidic devices
Testing involved healthy young adults as volunteers during
normal physical activities, and the volunteer subjects gave
informed consent. The skin was wiped with alcohol to clean
grease and ash to form strong adhesion to the device. Low-
intensity running was carried out at an ambient temperature
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of 35 °C. A volume of water was imbibed during the exercise,
and the sweat collection continued until the microfluidic
device was completely filled with sweat.
Conflicts of interest
The authors declare no competing financial interest.
Acknowledgements
This research was supported by the National Natural Science
Foundation of China (11872326 and 11702236), Natural Science
Foundation of Hunan Province (2018JJ2379 and 2018JJ2396),
Scientific Research Fund (18B089) of Hunan Provincial
Education Department, and Open Fund from Institute of
Flexible Electronics Technology of THU (2019KF1102). H. C.
also acknowledges the support from Pennsylvania State
University and American Chemical Society Petroleum Research
Fund (59021-DNI7). The helpful discussion with W. Xia at Xi'an
Jiaotong University is also acknowledged.
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