Article

Cite This: Anal. Chem. 2017, 89, 13160−13166 pubs.acs.org/ac

Paper−Plastic Hybrid Microfluidic Device for Smartphone-Based

Colorimetric Analysis of Urine
Uddin M. Jalal, Gyeong Jun Jin, and Joon S. Shim*
Bio IT Convergence Laboratory, Department of Electronic Convergence Engineering, Kwangwoon University, Seoul 139-701,
Republic of Korea
*
S Supporting Information

ABSTRACT: In this work, a disposable paper−plastic hybrid

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microfluidic lab-on-a-chip (LOC) has been developed and

successfully applied for the colorimetric measurement of urine
by the smartphone-based optical platform using a “Urine-
Analysis” Android app. The developed device was cost-
effectively implemented as a stand-alone hybrid LOC by
incorporating the paper-based conventional reagent test strip
inside the plastic-based LOC microchannel. The LOC device
quantitatively investigated the small volume (40 μL) of urine
analytes for the colorimetric reaction of glucose, protein, pH,
and red blood cell (RBC) in integration with the finger-actuating micropump. On the basis of our experiments, the conventional
urine strip showed large deviation as the reaction time goes by, because dipping the strip sensor in a bottle of urine could not
control the reaction volume. By integrating the strip sensor in the LOC device for urine analysis, our device significantly
improves the time-dependent inconstancy of the conventional dipstick-based urine strip, and the smartphone app used for image
analysis enhances the visual assessment of the test strip, which is a major user concern for the colorimetric analysis in point-of-
care (POC) applications. As a result, the user-friendly LOC, which is successfully implemented in a disposable format with the
smartphone-based optical platform, may be applicable as an effective tool for rapid and qualitative POC urinalysis.

U rine, like most waste products from the human body,

contains immense physiological information. Urinalysis
using reproducible standard procedures could provide an
effective tool to diagnose a number of human diseases and is
influential for the diagnosis of urinary tract infections and renal
disorders, malignancy, cardiovascular diseases, and metabolic
malfunctioning, such as diabetes or liver disease.1−4 A diseased
person can be identified by unusual or abnormal behavior of
the urine, which can be in variety of clarity, colors, smells, and
specific gravity.2,5 Ideal urinalysis procedures are cheap, require
limited technical support, and enable quick diagnosis with
significant accuracy.2,6 Dipstick or diagnostic reagent strips are
the first commercial lateral flow assay, which are commonly
used in urine analysis,7−10 and solely rely on the colorimetric
changes of reagent pads. The reagent pads change their color
on reaction with the urine analytes. Thus, the intensity of the
color change of the reagent pads corresponds to the
concentration of the analytes present in the urine specimen.10,11
Finally, the urine analytes reacted colored pads are matched
with a reference color chart that consists of a number of color
blocks, each block implying a certain concentration of the
analyzed analyte. Since the interpretation of the colorimetric
changes potentially suffers from observer variability, the
assessment is notably imprecise, labor-consuming, and a
subjective issue for point-of-care (POC) clinical applica-
tions,12,13 although the use of dipstick reagent pads is user-
convenient and cheap, with necessary compactness and
portability.
Highly sensitive automated dipstick readers have been
introduced to address the limitations of colorimetric assessment
using dipstick reagent pads in urinalysis, overwhelming
observer variability.14,15 The use of electrical readers has also
been suggested for the qualitative detection of a colorimetric
dipstick.16 Such automated or electrical dipstick readers for
urine analysis are inconvenient for resource-limited environ-
ments, because of their requirement of technical skill and
relatively high cost.17
Recently, smartphone-based lab-on-a-chip (LOC) devices
have been reported for colorimetric analysis, affording a
number of portable, and user-friendly, POC diagnostic
tools.18−23 Integration of a smartphone-based platform with
an LOC device for POC testing greatly reduces the diagnostic
cost, along with minimal instrumentation.24−29 The use of a
smartphone-embedded high-resolution camera can provide
rapid optical detection of bodily fluid biomarkers, both in the
ambient and in-house optical chamber.30−32 The external
housing unit that requires additional optical elements, including
a light source for imaging, such as a light-emitting diode (LED),
light diffuser, and optical chamber, provides more precise and
accurate on-site screening of liquid analytes. 18,33,34 A
smartphone app with the necessary algorithm conclusively
Received: July 5, 2017
Accepted: November 13, 2017
Published: November 13, 2017

© 2017 American Chemical Society 13160 DOI: 10.1021/acs.analchem.7b02612

Anal. Chem. 2017, 89, 13160−13166
Analytical Chemistry
Figure 1. (a) Conceptual scheme of a hybrid lab-on-a-chip (LOC) device made of patterned polycarbonate (PC) sheet for urinalysis, (b) urine
solution inside a cup, and (c and d) its operational steps. (c) Finger force is applied to initiate negative pressure to move sample solution into the
LOC device chamber, and (d) the solution flows into the device chamber to react with the reagent pads.
facilitates rapid quantification of the smartphone camera-
captured image of the liquid analyte.
In the conventional dipstick method, the dipping of a strip
sensor in a bottle of urine could not control the reaction
volume and results in the time-dependent variation for the
colorimetric assessment of the conventional test strip. To
address the issues experienced in the conventional method, this
work demonstrates a cost-effective, single-channel hybrid LOC
device integrated with an on-chip poly(dimethylsiloxane)
(PDMS) micropump for analyzing artificial urine analytes,
such as, glucose, protein, pH, and red blood cells (RBC), the
most common urine analytes reported for a number of diseases.
In the formation, an array of commercial paper-based reagent
test pads was embedded across the microchannel of a plastic-
made microfluidic device to form a paper and plastic based
hybrid LOC device. Since the microchannel working as reaction
chamber has a predefined volume, it contained the specific
volume of urine sample. So, in every measurement, the certain
volume of sample reacted with the test pads. The micropump
integrated with the LOC device enables the loading of sample
volume into the microchannel. Since the accuracy of the
colorimetric measurement is conceivably influenced by the
optical uniformity during image processing,25,30,35 an imaging
box made of white acrylic poly(methyl methacrylate) (PMMA)
sheet, reported previously by our group,18 has been integrated
with the developed platform. The imaging box including PDMS
light diffuser evenly distributes the camera flash, while
maintaining a consistent distance between the camera lens
and target reagent pads.
■
PRINCIPLES BEHIND THE DEVELOPED DEVICE,
OPTICAL PLATFORM, AND COLORIMETRIC
ANALYSIS
Device Principle. In conventional urinalysis, since the
sensitivity of the colorimetric analysis relies on the homoge-
neity of color of the urine analyte reacted reagent pads, the
accuracy in quantitative analysis has always been an inclusive
issue. For simple and quantitative colorimetric urinalysis,
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glucose, protein, pH, and RBC, the most common urine
analytes, have been considered using the hybrid LOC device
shown in conceptual view in Figure 1, parts a and b. The device
accommodates a paper-based reagent strip embedded into the
microchannel of a polycarbonate (PC) plastic-material-made
LOC device. The device used for the experiment was designed
as a single microfluidic channel structure with a single inlet for
drawing sample solution into the device reaction chamber and
an outlet connected to an elastic PDMS micropump. The
disposable PDMS micropump, which is outlined in Figure 1c,
initiates negative pressure inside the microchannel of the
device, to demonstrate the flow of sample analyte for reaction
with the embedded reagent pads. When a finger is pressed onto
the PDMS micropump, shrinking of the PDMS pump
compresses the unoccupied zone under the pump, and the
LOC device inlet is dipped into a sample solution container, as
in Figure 1b. When the finger is released, as shown in Figure
1d, negative pressure originated inside the microchannel of the
device drives the sample analytes through the LOC micro-
channel that serves as a reaction chamber. Eventually, the
smartphone camera senses the colorimetric change of the
reagent pads that have reacted with the analytes.
Optical Platform. The proposed hybrid LOC platform can
be utilized for the analysis of equal and controlled volume of
sample, fixing the test pads positions in a specific location for
image processing. Since image processing using a smartphone
camera is largely affected by the color rendering index (CRI)
values, uniform light needs to be provided.36,37 Hence, the
diagnostic LOC platform includes an imaging box of white
acrylic PMMA sheet, which is equipped with a light diffuser, as
previously described.18 The imaging box provides a reprodu-
cible test platform by unifying the ambient lighting conditions.
When the image of the test pads is captured inside the white
box under the flashlight, the flashlight transmitted through the
diffuser equally disperses on the test pads, and also the light
reflects from the white acrylic surface, thereby reducing the loss
of illumination, to avoid optical variations during image
processing.
DOI: 10.1021/acs.analchem.7b02612
Anal. Chem. 2017, 89, 13160−13166
Analytical Chemistry
Colorimetric Algorithm for the Conversion of RGB
Values to Analytical Hue Values. RGB (red, green, blue)
colors are the three major determinants of a smartphone
camera.12,32 The images that are sensed by the smartphone are
digitized into RGB color coordinates and show non-monotonic
behavior with light wavelength and intensity.38,39 Therefore, the
conversion of RGB data extracted from the smartphone
camera-captured image to other color spaces that correspond
to the color spectra of the analyzing strips is suggested.40
Regardless of the optical variation, as previously reported,41,42
the hue is more perceptual than the RGB values for the
quantification of the colorimetric reagent strip.
The conversion of RGB values of the smartphone camera-
captured image to their corresponding hue values is executed in
two consecutive steps. Initially, the nonlinear rgb pixels counted
from the image are linearized to estimate their RGB values
following the eq 1, which follow the mathematical relationship
of eq 2 to result in the respective hue value of the image.43−45
Finally, a “UrineAnalysis” app for the Android platform has
been developed to obtain the hue value from the RGB data
ranging from 0 to 255 based on the algorithms using eqs 1 and
2:
r g b
R= × 100, G = × 100, B = × 100
r+g+b r+g+b r+g+b
(1)
⎡ ⎤
−1⎢ 0.5{(R − G) + (R − B)} ⎥ for π ≥ H ≥ 0(G > B)
H = cos
⎢⎣ {(R − G)2 + (R − B)(G − B)} ⎥⎦
(2)
■
MATERIALS AND METHODS
Sample Preparation. “URiSCAN” reagent strips provided
by YD-Diagnosis, Korea were used to fabricate the hybrid
microfluidic LOC device for urinalysis. The chemicals for the
preparation of the artificial urine specimen were dextrose
powder (Streck, U.S.A.), albumin solution (Comscience Ltd.,
Korea), and pH buffer solutions (Samchun, Korea). Urinary
solutions of glucose and protein of different concentrations
were prepared using DI water; solution of different RBCs/μL of
sample, which was collected from a healthy donor in a hospital
following the local regulations, was prepared using phosphate
buffer saline (PBS). The concentrations associated with the
analysis were 0−350 mg/dL for glucose, 0−2000 mg/dL for
protein, 5.25−7.5 for pH, and 0−280 RBC/μL of solution for
red blood cells, which are in the typical physiological ranges
found in actual urine for a wide variety of screening issues.
These concentrations of glucose, protein, and RBC were
obtained through serial dilution of their standard solutions.
Device Fabrication. The hybrid POC testing platform
developed for urinalysis comprises (i) an array of commercial
URiSCAN reagent test pads embedded across the micro-
channel of the microfluidic hybrid LOC device, (ii) a white
acrylic imaging box, and (iii) a smartphone with the developed
Android “UrineAnalysis” app for the analysis of captured image
of the colorimetric test strip inside the microchannel of the
LOC device.
To fabricate the LOC device for urine analysis, the
technologically sensitive lithography process has been replaced
by a low-cost hot-embossing technique (Qmesys Corp., Korea),
using a laser cutter (C30, Coryart Inc., Korea) for patterning of
the microchannel of the device. Figure 2a demonstrates the
fabricated LOC device of 70 mm × 20 mm × 4 mm (length ×
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Figure 2. (a) Fabricated hybrid LOC device with PDMS micropump,
(b) insertion of urine analyte into the reagent chambers of the LOC
device by the elastic restoration force of a PDMS micropump, (c) a
smartphone on the top of a white acrylic imaging box, and (d) the
disposable hybrid LOC device inside the acrylic box for imaging. The
scale bars equal 1 cm.
width × height) dimensions, which includes two layers of PC
sheet with corresponding functional steps. One sheet of PC was
patterned as a microchannel with channel length, width, and
depth of 60 mm, 2 mm, and 200 μm, respectively, using the
laser cutter (C30, Coryart Inc., Korea), and also, four reaction
chambers of 5 mm2 area with a depth of 2 mm for the
accommodation of reagent pads were patterned across the
microchannel as in Figure 2a. Subsequently, the PC substrate
with patterned channel was thermally bonded on the blank PC
substrate to form the microfluidic channel using a hot press
with applied pressure of 2.5 bar at 140 °C for 10 min, and an
outlet hole of 1 mm diameter was drilled in the blank PC
substrate.
To prepare the disposable hybrid LOC platform, the reagent
strip including four different reagent pads as sensing area was
attached to the microfluidic channel of the LOC device through
the vacant sidewall of the device, and the sidewall was then
sealed with epoxy polymer. The elastic PDMS micropump was
fabricated by replica molding technique as reported pre-
viously39 and attached to the outlet of the LOC device, as
shown in Figure 2a, to load sample inside the microchannel.
The standard elasticity of the PDMS micropump was
confirmed by the mixing ratio of curing agent and Sylgard
184 (Dupont Inc., U.S.A.) of 1:14 and pouring the PDMS
mixture into a hemispherical mold, which was then cured at 70
°C for 4 h and shaped accordingly to prepare the micropump.
The fixation of the paper-based four reagent test pads into the
LOC device provides a number of measurement facilities over
the conventional dipstick methods, which have been
summarized as Table S1 in the Supporting Information.
The finger pressing and releasing from the PDMS micro-
pump that involve the flow of artificial urine analytes through
the device microchannel, as described in the Device Principle
section, are shown in Figure 2b. Figure 2c shows a smartphone
on the top of the white acrylic imaging box for imaging of the
LOC device. While the urine analytes interact with test strip
inside the microchannel, and colorimetric changes occur in the
reagent pads, the LOC device is positioned inside the imaging
DOI: 10.1021/acs.analchem.7b02612
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Analytical Chemistry Article

Figure 3. Measurement of hue value (a) varying the reaction time of the test strip into glucose solution of 50 mg/dL and (b) varying the removal
time of solution from the test strip while taking out from the glucose solution of 50 mg/dL in the conventional dipstick method.

Figure 4. Measurement of hue value with respect to sample volume of (a) 50 mg/dL glucose, (b) 300 mg/dL protein, (c) pH 5.5, and (d) 150
RBC/μL of solution using the developed LOC device.

box using a sample holder as shown in Figure 2d for imaging
with the smartphone.
Image Acquisition and Colorimetric Analysis of the
Image Using the Developed App. To quantify the hue
value from the colorimetric reaction, the LOC device including
the test pads is placed inside the imaging box, as in Figure 2d.
Then, a smartphone placed on top of the box captures the
image of the test pads, and the “UrineAnalysis” Android app
processes the images to extract the colorimetric information as
hue value following the functional steps as given in Figure S2 of
the Supporting Information.
been embedded into the LOC chip, includes test pads for the
measurement of urinary glucose, protein, pH, and RBC.
Limitations of Conventional Dipstick-Based Urinaly-
sis. Conventionally, urinalysis using the URiSCAN dipstick is
executed by exposing the dipstick reagent pad inside the urine
solution, and clinical diagnosis is interpreted based on the visual
colorimetric change of the reagent pads. The test method
suggests (i) dipping the test strip for 1 s inside the urine
solution, (ii) removing the leftover solution from the strip using
soft tissue when the strip is taken out of the solution, and (iii)
then leaving the strip for 1 min, referred to as the reaction time,

■ RESULTS AND DISCUSSION

Urinalysis using dipstick methods refers to the analysis of the
chemical composition of urine. The analysis can be readily
carried out to quantify the presence of protein, glucose, pH,
ketones, blood RBC, and bilirubin/urobilinogen in urine.
Several suppliers include additional reagent pads onto the test
strip to determine the white blood cells (WBC), nitrite, and
specific gravity of urine. The URiSCAN test strip, which has
13163
before colorimetric analysis. The colorimetric analysis for the
determination of the certain physiological disorder is assessed
by matching the color change of a selective reagent pad in the
strip with a reference color chart. But the entire process may
significantly suffer from the variability of dipping time and the
solution removal time from the strip. To inspect these time-
dependent issues for the regular dipstick method, studies have
been carried out as per the measurement guidelines from the
URiSCAN urine test assay suppliers.
DOI: 10.1021/acs.analchem.7b02612
Anal. Chem. 2017, 89, 13160−13166
Analytical Chemistry
Figure 3a shows the hue values for the glucose of 50 mg/dL
measured with varying dipping times and leftover sample
removal time of 1 s using the typical dipstick method. Since the
hue value corresponds to the same concentration of analyte, it
should be reliably constant for the entire dipping time. Figure
3a shows that the hue value decreases until the dipping time of
2 s, becomes relatively steady in the range of 2−4 s, and beyond
4 s eventually falls again. Until the dipping time of 2 s, the
decreasing hue value is most probably because of the increasing
number of molecules of the solution reacting with the reagent
pad. This means that the reagent pads absorb the proper
amount of urine solution within 2 s, whereas with longer
dipping time beyond 4 s, some molecules from the reagent pads
of the dipstick may get released into the glucose solution, which
may correspond to the unreliably varying hue values. But the
hue values were notably consistent from 2 to 4 s, thus
suggesting the dipstick test within this reaction phase. In
addition, when the test strip was taken out from the glucose
solution after 1 s of dipping time, the time taken to remove
solution by tissue was measured to study the difference
between the quick removal and the slow removal of urine.
Figure 3b shows that the hue values decrease with increasing
removal time, since the longer removal time may wash some
molecules away from the reagent pads, resulting in nonuniform
colorimetric changes on the pads. Thus, the sample volume
with processing time is influential to the colorimetric
quantification of urine analytes.
Optimization of Urine Sample Volume. Since the
regular dipstick method suffers from time-dependent variability
in terms of the dipping time and the solution removal time, an
LOC embedding the reagent pads inside its microchannel has
been proposed so that a certain amount of urine can be reacted
with the urine strip to address those issues. In addition, the
proposed device can improve the measurement inconstancy
ignoring the sample removal step during analysis. Since the
novelty of the developed hybrid LOC platform is the precisely
controlled volume of analyte used for the colorimetric
assessment, to optimize the volume of urine that must flow
into the microfluidic device, various volumes of urine sample
solution were flowed into the microfluidic device to measure
the change in the corresponding hue value.
Parts a−d of Figure 4 show the hue values in terms of the
controlled volume of glucose, protein, pH, and RBC of blood,
using the developed paper−plastic-based LOC hybrid device.
The results showed that the hue value was stable for more than
30 μL of the sample solution, which means that the sample
solution of more than 30 μL is required to entirely react with
the individual reagent pad. On the basis of the experimental
results, a micropump was made that can move 40 μL of urine
sample for the accurate and detailed colorimetric analysis of
urine.
Comparison of the Results of the LOC-Based Micro-
fluidic Chip and Conventional Dipstick Test. The key
achievement of the proposed LOC device over the dipstick
method is the comparably uniform color change of the test
pads while reacting with the urine analytes inside the
microchannel that function as reaction chamber of the device.
This is because of the defined volume of the microchannel
accommodating a precise amount of urine sample and the
certain volume of sample reacts with the test pads in each
measurement. A comparative data for the typical dipstick
method and the developed LOC hybrid platform, with the
variation of reaction time is shown in Figure 5.
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Figure 5. Measurement of hue value varying the reaction time of the
reagent strip in glucose of 50 mg/dL using both the dipstick method
(cyan blue data) and the developed LOC device (brown data).
This graph shows the change in hue value over time for the
same concentration of glucose. The result presents that, when
the sample is analyzed using the developed LOC device, since a
certain volume of urine reacts with the urine strip inside the
microchannel, the color change as well as the change in hue
value with the reaction time is small, and the value is almost
similar beyond 90 s. However, in the case of the dipstick
method, hue value with systematic variation of reaction time
with 1 s leftover sample removal time was measured, where the
change in the hue value over time is relatively large. Thus, the
developed LOC device shows less impact on reaction time with
improved reproducibility and accuracy of urinalysis. Also, as
there is no leftover sample removal issue in the LOC-based
urinalysis, the inaccuracy of the colorimetric measurement
experienced in the dipstick method as shown in Figure 3b is
entirely avoided in the proposed LOC platform.
Detailed Analysis of Urine Analytes. After the
demonstration of a proof of concept for the optimization of
time variable inaccuracy experienced in the dipstick method,
and the advantages of volume-controlled measurement using
the developed LOC device, the analysis technique has been
transferred to the detailed measurement of glucose, protein,
pH, and RBC, and the corresponding hue values are shown in
Figure 6.
The figure corresponds to the hue values for the glucose of
0−350 mg/dL, protein of 0−2000 mg/dL, pH of 5.25−7.5, and
red blood cells of 0−280 RBC/μL of solution. The data and the
error bars in the figures correspond to the respective mean and
relative standard deviation. With increasing concentration, the
hue value increases linearly for protein, pH, and RBC, and
decreases linearly for glucose. As seen from the Figure 6a, the
hue value for glucose beyond 300 mg/dL is small and constant,
which is because of the almost equal color changes of the
reagent strips at higher concentration above 300 mg/dL of
glucose. The measurement covers a wide range of concen-
trations that are in the clinical detection range for glucose,
protein, RBC, and pH.30,40,46 Thus, the developed platform
allows for a greater dynamic range of quantitative measure-
ment of glucose, pH, protein, and occult blood with increased
sensitivity.
Paper-based dipstick tools are promising as they are
implemented cost-effectively for the resource-limited develop-
ing societies in many countries.47 But their practical
applicability suffers from time-dependent measurement in-
accuracy and observer variability in qualitative colorimetric
changes of the test pads. Assembling of test pads across the
microchannel in a plastic-made LOC device qualitatively
DOI: 10.1021/acs.analchem.7b02612
Anal. Chem. 2017, 89, 13160−13166
Analytical Chemistry Article

Figure 6. Measurement of the hue value for (a) glucose, (b) protein, (c) pH, and (d) RBC of different concentrations.

improves the measurement variation requiring minimal volume

of analytes. Optimization of color balancing in ambient lighting
■
*
ASSOCIATED CONTENT
S Supporting Information
conditions using a smartphone camera, which is a major The Supporting Information is available free of charge on the
challenge in colorimetric assays, has already been addressed by ACS Publications website at DOI: 10.1021/acs.anal-
different approaches using an external opto-mechanical housing chem.7b02612.
unit along with a smartphone camera including batteries, LED Advantages of the proposed paper−plastic hybrid
arrays, and external lenses.48−51 In this work, a unique optical microfluidic LOC, image acquisition and colorimetric
box equipped with simplified PDMS light diffuser previously analysis of the image, reaction phenomena that happen
on the reagent pad, and analysis of glucose in real urine
reported by our group18 has been adopted, eliminating the and PBS buffer (PDF)

■
design complicated LED arrays and highly priced external
lenses for the colorimetric analysis of urine assays. The AUTHOR INFORMATION
dimension of the white-acrylic-made optical box is phone-
Corresponding Author
specific to fix the optimal distance between the smartphone *Phone: +82-10-4121-3075. E-mail: shim@kw.ac.kr.
camera and target sample that could be revised with minimal
ORCID
redesigning for different specifications of the smartphones. Uddin M. Jalal: 0000-0003-2533-8019

■ CONCLUSIONS
In this work, a unique low-priced hybrid microfluidic LOC
device combined with a smartphone-based optical platform has
been proposed for the colorimetric detection of urinary
biomarkers. The hybrid device successfully demonstrated the
precisely controlled volume of sample solution for a fixed
colorimetric reaction time inside the device microchannel,
thereby reducing time-dependent measurement inaccuracy.
The utilization of the smartphone app as an optical reader
and image processor made the colorimetric urinalysis
convenient, reducing the observer and measurement variability
that arise in the typical colorimetric POC diagnosis. Thus, the
developed smartphone-based hybrid LOC platform can be
widely applicable for other colorimetric assays in clinical
applications.
Notes
The authors declare no competing financial interest.
■ ACKNOWLEDGMENTS
The authors gratefully acknowledge the support for this work
from a Kwangwoon Research Grant of 2017. This research was
also partly supported by the Technological Innovation R&D
program of the SMBA (S2498668) and the Basic Science
Research Program through the National Research Foundation
of Korea (NRF), funded by the Ministry of Education (no.
NRF-2015R1D1A1A01060369).
■ REFERENCES
(1) Coppens, A.; Speeckaert, M.; Delanghe, J. Acta. Clin. Belg. 2010,
65, 182−189.
(2) Devillé, W. L. J. M.; Yzermans, J. C.; van Duijn, N. P.; Bezemer,
P. D.; van der Windt, D. A.; Bouter, L. M. BMC Urol. 2004, 4 (1), 4−
17.

13165 DOI: 10.1021/acs.analchem.7b02612

Anal. Chem. 2017, 89, 13160−13166
Analytical Chemistry Article

(3) Kanegaye, K. T.; Jacob, J. M.; Malicki, D. Pediatrics 2014, 134 (35) García, A.; Erenas, M. M.; Marinetto, E. D.; Abad, C. A.; de
(3), 523−529. Orbe-Paya, I.; Palma, A. J.; Capitan-Vallvey, L. F. Sens. Actuators, B
(4) Simerville, J. A.; Maxted, W. C.; Pahira, J. J. Am. Fam. Physician. 2011, 156, 350−359.
2005, 71 (6), 1153−1162. (36) Lopez-Ruiz, N.; Curto, V. F.; Erenas, M. M.; Benito-Lopez, F.;
(5) McPherson, R. A.; Pincus, M. R. Henry’s Clinical Diagnosis and Diamond, D.; Palma, A. J.; Capitan-Vallvey, L. F. Anal. Chem. 2014,
Management by Laboratory Method; Elsevier/Saunders: Philadelphia, 86, 9554−62.
PA, 2011. (37) Wang, R.; Prabhakar, A.; Iglesias, R. A.; Xian, X.; Shan, X.;
(6) Chenari, M. R.; Gooran, S.; Zarghami, A.; Fazeli, F. OJU 2012, 2, Tsow, F.; Forzani, E. S.; Tao, N. IEEE Sens. J. 2012, 12, 1529−1535.
227−231. (38) Murdock, R. C.; Shen, L.; Griffin, D. K.; Kelley-Loughnane, N.;
(7) Abirami, K.; Tiwari, S. C. J. Ind. Acad. Clin. Med. 2001, 2 (1), 39− Papautsky, I.; Hagen, J. A. Anal. Chem. 2013, 85, 11634−11642.
50. (39) Lee, H. C. Introduction to Color Imaging Science; Cambridge
(8) Delanghe, J.; Speeckaert, M. Biochemia. Medica. 2014, 24 (1), University Press: Cambridge, U.K., 2005.
89−104. (40) Oncescu, V.; O’Dell, D.; Erickson, D. Lab Chip 2013, 13, 3232−
(9) Ko, K.; Kwon, M.-J.; Ryu, S.; Woo, H.-T.; Park, H. J. Clin. Lab. 38.
Anal. 2016, 30 (5), 424−430. (41) Chang, B. Y. Bull. Korean Chem. Soc. 2012, 33, 549−52.
(10) Pugia, M. J. Lab. Med. 2000, 31 (2), 92−96. (42) Cantrell, K.; Erenas, M. M.; de Orbe-Payá, I.; Capitán-Vallvey,
(11) Mundt, L. A.; Shanahan, K. Graff’s Textbook of Routine Urinalysis L. F. Anal. Chem. 2010, 82 (2), 531−542.
and Body Fluids; Wolters Kluwer/Lippincott Williams & Wilkins (43) Khan, M.; Jamil, A.; Haleem, F.; Muhammad, Z. A. Middle-East
Health: Philadelphia, PA, 2016. J. Sci. Res. 2014, 22 (5), 647−654.
(12) Hong, J. I.; Chang, B. Y. Lab Chip 2014, 14, 1725−1732. (44) Marshall, W. J.; Bangert, S. K. Clinical Biochemistry: Metabolic
(13) Memişoğullar, R.; Yüksel, H.; Yildirim, H. A.; Yavuz, Ö . Eur. J. and Clinical Aspects; Churchill Livingstone: Edinburgh, U.K., 1995.
(45) Kim, H.; Awofeso, O.; Choi, S.; Jung, Y.; Bae, E. Appl. Opt.
Gen. Med. 2010, 7 (2), 174−178.
2017, 56 (1), 84−92.
(14) De Silva, D. A.; Halstead, A. C.; Côté, A. M.; Sabr, Y.; von
(46) Im, S. B.; Kim, S. C.; Shim, J. S. Anal. Bioanal. Chem. 2016, 408,
Dadelszen, P.; Magee, L. A. J. Obstet. Gynaecol. Can. 2014, 36 (7),
1391−1397.
605−612. (47) Vashist, S. K.; Luppa, P. B.; Yeo, L. Y.; Ozcan, A.; Luong, J. H.
(15) Langlois, M. R.; Delanghe, J. R.; Steyaert, S. R.; Everaert, K. C.; T. Trends Biotechnol. 2015, 33, 692−705.
De Buyzere, M. L. Clin. Chem. 1999, 45 (1), 118−122. (48) Mudanyali, O.; Dimitrov, S.; Sikora, U.; Padmanabhan, S.;
(16) Jeong, S. G.; Kim, J.; Nam, J. O.; Song, Y. S.; Lee, C. S. Int. Navruz, I.; Ozcan, A. Lab Chip 2012, 12, 2678−2686.
Neurourol. J. 2013, 17 (4), 155−161. (49) Dutta, S.; Sarma, D.; Patel, A.; Nath, P. IEEE Photonics Technol.
(17) Lee, D.-S.; Jeon, B. G.; Ihm, C.; Park, J.-K. Lab Chip 2011, 11, Lett. 2015, 27 (22), 2363−2366.
120−126. (50) Dutta, S.; Saikia, K.; Nath, P. RSC Adv. 2016, 6, 21871−21880.
(18) Kim, S. C.; Jalal, U. M.; Im, S. B.; Ko, S.; Shim, J. S. Sens. (51) Kim, S. D.; Koo, Y.; Yun, Y. Sensors 2017, 17, 1604.
Actuators, B 2017, 239, 52−59.
(19) Chen, A.; Wang, R.; Bever, C. R. S.; Xing, S.; Hammock, B. D.;
Pan, T. Biomicrofluidics 2014, 8, 064101−064111.
(20) Barbosa, A. I.; Gehlot, P.; Sidapra, K.; Edwards, A. D.; Reis, N.
M. Biosens. Bioelectron. 2015, 70, 5−14.
(21) Roda, A.; Michelini, E.; Cevenini, L.; Calabria, D.; Calabretta,
M. M.; Simoni, P. Anal. Chem. 2014, 86, 7299−7304.
(22) You, D. J.; Park, T. S.; Yoon, J.-Y. Biosens. Bioelectron. 2013, 40,
180−185.
(23) Kim, S.; Cho, D.; Kim, J.; Kim, M.; Youn, S.; Jang, J. E.; Je, M.;
Lee, D. H.; Lee, B.; Farkas, D. L.; Hwang, J. Y. Biomed. Opt. Express
2016, 7 (12), 5294−5307.
(24) Pierce, M. C.; Weigum, S. E.; Jaslove, J. M.; Richards-Kortum,
R.; Tkaczyk, T. S. Ann. Biomed. Eng. 2014, 42, 231−240.
(25) Shen, L.; Hagen, J. A.; Papautsky, I. Lab Chip 2012, 12, 4240−
4243.
(26) Weigl, B.; Domingo, G.; Labarre, P.; Gerlach, J. Lab Chip 2008,
8 (12), 1999−2014.
(27) Zhu, H.; Isikman, S. O.; Mudanyali, O.; Greenbaum, A.; Ozcan,
A. Lab Chip 2013, 13, 51−67.
(28) Jalal, U. M.; Kim, S. C.; Shim, J. S. Biomed. Opt. Express 2017, 8
(7), 3317−3328.
(29) Vashist, S. K.; van Oordt, T.; Schneider, E. M.; Zengerle, R.; von
Stetten, F.; Luong, J. H. T. Biosens. Bioelectron. 2015, 67, 248−255.
(30) Yetisen, A. K.; Martinez-Hurtado, J. L.; Garcia-Melendrez, A.; da
Cruz Vasconcellos, F.; Lowe, C. R. Sens. Actuators, B 2014, 196, 156−
160.
(31) Lee, S.; Oncescu, V.; Mancuso, M.; Mehta, S.; Erickson, D. Lab
Chip 2014, 14, 1437−1442.
(32) Koesdjojo, M. T.; Pengpumkiat, S.; Wu, Y.; Boonloed, A.;
Huynh, D.; Remcho, T. P.; Remcho, V. T. J. Chem. Educ. 2015, 92 (4),
737−741.
(33) Jung, Y.; Kim, J.; Awofeso, O.; Kim, H.; Regnier, F.; Bae, E.
Appl. Opt. 2015, 54 (31), 9183−9189.
(34) Wang, Y.; Li, Y.; Bao, X.; Han, J.; Xia, J.; Tian, X.; Ni, L. Talanta
2016, 160, 194−204.

13166 DOI: 10.1021/acs.analchem.7b02612

Anal. Chem. 2017, 89, 13160−13166