Materials Chemistry and Physics 173 (2016) 285e290

Contents lists available at ScienceDirect

Materials Chemistry and Physics

journal homepage: www.elsevier.com/locate/matchemphys

A bioinspired color-changing polystyrene microarray as a rapid

qualitative sensor for methanol and ethanol
Wen-Kai Kuo a, Hsueh-Ping Weng a, Jyun-Jheng Hsu a, Hsin Her Yu b, *
a
Graduate Institute of Electro-Optical and Materials Science, National Formosa University, 64 Wenhua Road, Huwei, Yunlin 63208, Taiwan
b
Department of Biotechnology, National Formosa University, 64 Wenhua Road, Huwei, Yunlin 63208, Taiwan

h i g h l i g h t s g r a p h i c a l a b s t r a c t

Opal-like array of polystyrene (PS)

microspheres is synthesized by self-
assembly.

This periodic PS array is used as a
rapid sensor for methanol and
ethanol.

Solvents are detected by routes of
reflection coordinates in chromaticity
diagram.

They are also detected directly by
naked eye based on change in color
of sensor.

The color change is irreversible for
methanol but reversible for ethanol.

a r t i c l e i n f o a b s t r a c t

Article history: Polystyrene (PS) microspheres were synthesized by emulsifier-free emulsion polymerization and ar-
Received 10 March 2015 ranged in an array of closely packed, opal-like photonic crystals by slow self-assembly through dip-
Received in revised form coating. This periodic array of PS microspheres was then employed as a rapid qualitative sensor for
1 February 2016
methanol and ethanol. Both solvents could be detected rapidly based on the routes of their reflection
Accepted 4 February 2016
Available online 12 February 2016
coordinates in the chromaticity diagram or directly by the naked eye on the basis of the change in color
within 1 min once a solvent sample had been placed on the PS photochromic sensor. This opal-like PS
sensor can thus not only be employed as a rapid sensor for methanol and ethanol but can also be used as
Keywords:
Nanostructures
a powerful tool for the fast screening of illicit drugs and toxic chemicals during forensic investigations.
Polymers © 2016 Elsevier B.V. All rights reserved.
Surfaces
Visible and ultraviolet spectrometers

1. Introduction Pachyrhynchus argus have properties similar to those of photonic

In 2003, Parker et al. [1] reported that the elytra of the beetle
Abbreviations: DRS, dynamic reflection spectra; FESEM, field-emission scanning
electron microscopy; PS, polystyrene.
* Corresponding author.
E-mail addresses: wkkuo@nfu.edu.tw (W.-K. Kuo), sherry.weng7949@gmail.com
(H.-P. Weng), k88520x@gmail.com (J.-J. Hsu), hhyu@nfu.edu.tw (H.H. Yu).
crystals and have a structure similar to that of opal, exhibiting a
uniform metallic luster from all viewing angles. Opal is not a perfect
photonic crystal but is composed of many smaller crystals with
different orientations, such that it can reflect light of different fre-
quencies and produce beautiful structural colors. Owing to the
interesting colors and related properties exhibited by biological
structures, bioinspired photonic crystals are being explored for use
in optical engineering [2], electronic microwave communications

http://dx.doi.org/10.1016/j.matchemphys.2016.02.014
0254-0584/© 2016 Elsevier B.V. All rights reserved.
286 W.-K. Kuo et al. / Materials Chemistry and Physics 173 (2016) 285e290
[3], biomedical applications [4], and agricultural biotechnology [5].
The development of a rapid sensor for methanol and ethanol
would be highly beneficial. High-performance liquid chromatog-
raphy [6], gas chromatography [7], fuchsine sulfurous acid color-
imetry [8], and chromotropic acid colorimetry [9] are the methods
used traditionally to detect methanol. However, the pretreatments
involved in these methods are time consuming and costly, and the
testing procedures themselves are tedious.
In the past years, photonic sensors for liquid chemical species
have been realized based on the shift in the balanced reflection
peak and the change in their structural color. When a solvent in-
filtrates the voids of an opal solid, the effective refractive index of
the entire colloidal crystal increases, because the air is replaced by
the solvent, leading to a red shift in the reflection. The reflection
wavelength exhibits a linear relationship with the solvent's
refractive index, so that the simulated straight line can be used to
distinguish between different solvents. There are numerous situa-
tions where chemical sensors that are small, portable, easy to
handle, and capable of on-line monitoring a chemical species are
needed; these should ideally have a visual (i.e., visible to the naked
eye) readout. Sensors based on photonic crystals are promising in
this respect, particularly because they are not affected by electro-
magnetic fields [10,11].
It has been reported that, when the wings of the butterfly
Morpho menelaus, which are blue, are wetted with ethanol, the
resulting change in their refractive index causes them to turn green.
As the ethanol evaporates, the wings gradually turn blue again [12].
Inspired by the idea of developing photonic sensors similar to these
wings, in this study, we fabricated a rapid, qualitative sensor for
methanol and ethanol based on an array of polystyrene (PS) pho-
tonic crystals.
2. Materials and methods
2.1. Computing required PS particle size
Green light can be observed readily by the naked eye. If one
wishes to ensure that green light (l ¼ 495e510 nm) is reflected by
an array of PS photonic crystals, one can use Bragg and Snell's laws
to calculate the required crystal size [13,14]:
l
D ¼ qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
 
2
2 3 n  sin2 q
2
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
n¼ n2sphere  0:74 þ n2air  0:26
where D is the size of the PS microspheres, l is the reflection
wavelength of the PS photonic crystal array (or the opal-like
structure), q is the angle of incidence of the light, and n is the
average refractive index of the PS photonic crystal array in Eq. (1). In
Eq. (2), nsphere is the refractive index of PS (n ¼ 1.600) and nair is the
refractive index of air (n ¼ 1.000). Based these equations, the size of
PS microspheres such that they reflected green light was found to
be 223e273 nm.
2.2. Fabrication of opal-structured PS sensor
The polymerization procedure used to form the array of PS
microspheres was as follows. A mixture of deionized water (90 ml),
styrene (10 ml), and 4-styrenesulfonic acid sodium salt (40 mg) was
placed in a four-necked flask equipped with a reflux condenser and
a mechanical stirrer. After the reaction mixture had been homo-
geneously mixed, its temperature was raised to 70  C. Next, a
deoxygenated aqueous solution of potassium persulfate (0.9 mg)
was added to it. The emulsion polymerization process was stopped
after 24 h. Owing to the hydrophobicity of the glass substrate sur-
face, an oxygen plasma treatment was necessary. The oxygen
plasma (PCD-150, All Real Technology Co., Ltd.) treatment was
performed at a pressure of 300 mTorr using a 20 sccm oxygen flow
and an electrical power of 100 W. Next, the PS suspension was
diluted with deionized water to a concentration of 20 wt%, and the
opal-structured PS microspheres were arranged on the hydrophilic
glass surface at a dip-coating rate of 1 mm/s (see Fig. 1) [15].
2.3. Characterization of sensor
The average diameter and size distribution of the PS micro-
spheres in water were determined using a Zetasizer system (Mal-
vern 3000HS, UK). The morphology of the opal-like PS structure
was examined with a field-emission scanning electron microscopy
(FESEM) system (JEOL JSM-6700F, Japan), which was operated at an
accelerating voltage of 15 kV in the secondary electron image mode.
The samples were coated with a thin layer of gold by vapor depo-
sition using a vacuum sputtering system (Sputter JEOL JFC-1100C)
prior to FESEM characterization. The reflectance spectra and the
routes in the Commission Internationale d'Eclairage (CIE)-1931
chromaticity diagram of the solvents tested were determined using
a variable-angle multifunctional optical characteristics measuring
system (HMT MF-630). This apparatus was equipped with a
barium-sulfate-coated standard integrating sphere and a halogen
light source. The HMT MF-630 is a high-resolution spectrometer
capable of measuring the absorbance, transmittance, and reflec-
tance of solid (film) samples for wavelengths of 300e1100 nm.
3. Results and discussion
3.1. Analysis of opal-like PS structure
3.1.1. Average size and size distribution of synthesized PS
microspheres
The average particle size of the synthesized PS microspheres
was 242 nm, as determined using the Zetasizer system. Their par-
ticle size distribution index was 0.006 (see Fig. 2), indicating that
the PS particles were very uniform in size.
(1) 3.1.2. Analysis of surface of opal-like PS structure
Fig. 3(a) and (b) show SEM images of the surface and the cross-
section, respectively, of the opal-like PS structure. Though a few
local defects and vacancies existed in the PS microarray, most of the
(2)
PS microspheres were uniformly dispersed and arranged in a
compact and ordered periodic structure. The stacking thickness of
the PS microspheres was approximately 3.63 mm (~15 layers), as
evaluated from the SEM image in Fig. 3(b). This orderly arrange-
ment of the PS microspheres was attributable to the balance be-
tween the van der Waals attraction and the electrostatic repulsion
between the microspheres at the air/suspension interface. In order
to improve the ethanol and methanol detection sensitivity of the PS
structure, the thickness of the PS microarray was optimized; the
optimal thickness was found to be approximately 3.63e4.84 mm
(i.e., 15 to 20 layers in thickness). The stacking of an excessive
number of PS layers can lead to cracking as well as inhomogeneities
in the resulting opal film [16].
The stacking thickness of the PS microspheres on the substrate
was the primary factor influencing the reflectance (R%). The PS
microspheres arranged on the substrate tended to be white, since
the PS suspension was a white emulsion. When the PS suspension
concentration was increased from 20 to 100 wt%, the stacking
thickness of the PS microspheres on the substrate increased. This
 W.-K. Kuo et al. / Materials Chemistry and Physics 173 (2016) 285e290
Fig. 1. Schematic of the process for preparing the polystyrene (PS) microarray.
Fig. 2. Particle size and size distribution of the polystyrene microspheres.
Fig. 3. Scanning electron microscopy images of the polystyrene microarray: (a) top view and (b) cross-sectional view.
decreased the amount of light scattered over the microspheres and
thus the R% value (see Ref. [15]). Therefore, in order to be able to
detect the change in the color of the microarray with the naked eye
directly, the substrate with a monolayer of the PS microspheres was
not considered.
The reproducibility of the PS microarray was affected by the
following factors. (1) Environmental interference: vibrations
should be avoided when preparing the opal films, since the PS
microarray is prepared by a slow self-assembly-based dip-drawing
method. Further, the dip-drawing rate is also an important
parameter to consider when fabricating the microarray. (2) Pre-
treatment of the glass substrate: The PS microspheres did not
attach readily to the glass substrate, owing to its hydrophobic
surface. Therefore, an oxygen plasma treatment was required. (3)
The quality of the PS suspension: the particle size distribution of
the synthesized PS microspheres should be very uniform.
Furthermore, the stacking thickness of the PS microspheres on the
substrate was determined primarily by the concentration of the PS
suspension.
287
3.2. Application of opal-like PS sensor
3.2.1. Detection of methanol and ethanol based on their routes in
CIE diagram
Considering the fact that methanol and ethanol are volatile, the
testing environment was kept at a constant temperature and hu-
midity by using an air conditioner, in order to eliminate any
possible interference from the environment during the sensing
process. Thus, we placed the samples and the variable-angle
multifunctional optical characteristics measuring system in a
rectangular acrylic box during the sensing tests. The spectra data
were recorded using an optical fiber spectrometer (Ocean Co., USA)
within 60 s after the methanol and ethanol samples (500 mL) had
been dropped onto the PS sensor; the testing conditions were the
following: temperature of 25  C and relative humidity of 45%.
During the sensing tests, the solvents infiltrated the interstices
between the microspheres.
The time-dependent reflectance spectrum and the route of the
reflection coordinates in the chromaticity diagram for methanol are
shown in Fig. 4(a) (left, right). Not only did the route of the
reflection positions not return to its original position after meth-
anol testing, but the end points in the CIE diagram were far away
from the start points. Further, the response of the sensor to ethanol
was very different from that to methanol (see Fig. 4(b) (left, right)).
It can be seen from Fig. 4(b) that the reflection coordinates in the
chromaticity diagram returned to their original positions after the
ethanol sample had evaporated completely.
The human eye can detect different wavelengths and can
reconstruct colors in a three-dimensional manner. However, the
concept of color can be divided into two parts: luminance and
chrominance. Mathematically, x and y are the coordinates of the
projector (x is the red component and y is the green component).
Using Grassmann's law, one can use the reflection spectral data to
determine the CIE coordinates [17,18]. Therefore, we could deter-
mine the sensor's responses to the two solvents by converting their
time-dependent spectra into the corresponding CIE coordinates.
288 W.-K. Kuo et al. / Materials Chemistry and Physics 173 (2016) 285e290
Fig. 4. (a, b-left) Time-dependent reflection spectra of the polystyrene microarray corresponding to methanol and ethanol, respectively. (a, b-right) Routes corresponding to (a, b-
left) in the CIE chromaticity diagram.
Fig. 5 (a and b) are the CIE routes for methanol and ethanol,
respectively, as determined over several detection runs; the start
and end points in the CIE diagram are represented by circles and
rhombic symbols, respectively. We found that the CIE routes for
methanol were near the center (CIE(x,y) at (0.33,0.33), that is, cor-
responding to the color white) of the CIE diagram; however, this
was not the case for ethanol. In addition, the end points associated
with the start points of each of the CIE routes were separated from
each other. On the other hand, each pair of end points and start
points of the CIE routes for ethanol was either very close or
exhibited overlapping (see Table 1). That is to say, we could observe
the difference between them visually. Further, both methanol and
ethanol could also be detected within 1 min after being placed on
the sensor on the basis of the change in the color of the sensor,
which could be seen with the naked eye. Fig. 6 shows the change in
the color at time intervals of 5 s after samples of methanol and
ethanol had been placed on the PS sensor. A few residual styrene
monomers still existed in the self-assembled array of PS micro-
spheres; these dissolved once the methanol penetrated into the
array. Therefore, the change in color was irreversible in the case of
methanol but reversible for ethanol. This phenomenon was very
similar to the change in color observed when the wings of the
butterfly M. menelaus are wetted with ethanol.
In addition, the time-dependent reflectance of the PS sensor
decreased when the sensor was exposed to methanol or ethanol for
3e9 s. This phenomenon can be explained by Eq. (3).
" #
ðn1  n2 Þ2
Rð%Þ ¼  100% (3)
ðn1 þ n2 Þ2
where n1 and n2 are the refractive indices of the two dielectric
materials, respectively (methanol and ethanol in this case). Further,
it is assumed that the angle of incidence is 0. The reflectance values
corresponding to the two materials (methanol and ethanol) as
determined by Eq. (3) are listed in Table 2. The refractive index of
the opal-like structure changed when methanol or ethanol infil-
trated the interstices between the PS microspheres, since the
dielectric material (air) in the interstices was replaced; this also
induced a change in the reflectance. The larger the difference in the
 W.-K. Kuo et al. / Materials Chemistry and Physics 173 (2016) 285e290 289

Fig. 5. Routes in the CIE diagram corresponding to several runs to detect (a) methanol and (b) ethanol. The insets are the local magnification of the corresponding diagrams.

Table 1
CIE chromaticity coordinates corresponding to methanol and ethanol.

Fig. 6. Time-dependent changes in the color of the polystyrene microarray in response to methanol and ethanol. (For interpretation of the references to color in this figure legend,
the reader is referred to the web version of this article.)
290 W.-K. Kuo et al. / Materials Chemistry and Physics 173 (2016) 285e290
Table 2
Reflectance (%) values of the polystyrene (PS) microarray for different materials in
the interstices of the microarray.
Dielectric 1 PS, (n1 ¼ 1.600)
Dielectric 2 Air Methanol
(n2) 1.000 1.329
Reflectance (%) 5.325 0.856
Note: n1 ¼ refractive index of PS; n2 ¼ refractive index of air, methanol, or ethanol.
refractive indices of the PS microspheres and the dielectric material
present in the interstices, the higher was the reflectance of the
opal-like structure. On the other hand, if the difference were to be
small, the reflectance of the opal-like structure would also be low
[19]. In addition, we observed that the reflectance of the opal-like
PS sensor increased gradually as the methanol or ethanol sample
placed on the sensor evaporated (see the sections of the time-
dependent reflection spectra in Fig. 4(a) and (b), left correspond-
ing to 36e60 s). Both solvents could be detected by the naked eye
on the basis of the Bragg diffractive shifts of the ordered microarray
or by converting their time-dependent reflectance spectra into the
corresponding CIE routes.
Zheng et al. [20] have shown that polydimethylsiloxane-based
photonic crystal arrays can be used to visually detect the pres-
ence of nonpolar solvents such as the hydrocarbon n-hexane.
Furthermore, similar studies performed on ethanol/methanol and
ethanol/water mixtures have suggested that there exists a virtually
linear relationship between the peak wavelength and the fraction
of ethanol in the ethanol/methanol (or water) mixture. This high-
lights the advantages of photonic-crystal-based sensing over
indicator-based methods.
However, traditional photonic sensors based on the change in
the balanced reflection of photonic structures can barely distin-
guish between organic solvents with similar refractive indices. In
this study, we developed a photonic sensing method based on the
dynamic reflection spectra (DRS) of photonic crystals for dis-
tinguishing between organic solvents with similar structures and
physical properties. By testing a suitable number of solvents, a
database of standard DRS patterns can be established, such that it
would be useful for identifying unknown solvents [10].
4. Conclusions
In this study, we developed a new method for the convenient,
rapid, and sensitive detection of methanol and ethanol based on the
change in the refractive index of an opal-like sensor consisting of an
array of PS microspheres. The refractive index of the sensor
changed when the solvent to be detected infiltrated the interstices
between the PS microspheres in the array. Thus, the fabricated PS
microarray sensor was similar to the wings of M. menelaus and
showed a rapid response. Once drops of methanol or ethanol were
placed on the PS sensor, the particular solvent could be detected
rapidly (within 1 min) from the route of its reflection in the CIE
diagram or on the basis of the change in the color of the sensor,
which could be observed visually. However, three crucial differ-
ences were observed during the detection of methanol and ethanol.
First, in the case of the ethanol, the route of the reflection returned
to the original reflection coordinates in the CIE diagram after the
ethanol sample had evaporated completely; this was not the case
for methanol. Next, the route in the case of methanol lay near the
center of the CIE diagram, while that of ethanol did not. Lastly, the
change in the color of the sensor in the case of methanol was
irreversible, as methanol dissolved the residual styrene monomers.
On the other hand, the response to ethanol was reversible. Thus,
this opal-like PS sensor can be used as a powerful tool for the fast
Ethanol screening of illicit drugs and toxic chemicals during forensic
1.362
0.645
investigations.
Conflict of interest
The authors declare no conflict of interest.
Acknowledgment
The authors gratefully acknowledge financial support from the
National Science Council of Taiwan (NSC 102-2221-E-150-052 &
MOST 103-2221-E-150-021).
References
[1] A.R. Parker, V.L. Welch, D. Driver, N. Martini, Structural colour: opal analogue
discovered in a weevil, Nature 426 (2003) 786e787.
[2] T. Suezaki, H. Yana, T. Hatayama, G.A. Ozin, T. Fuyuki, Photoconductivity in
inverse silicon opals enhanced by slow photon effect: yet another step to-
wards optically amplified silicon photonic crystal solar cells, Appl. Phys. Lett.
98 (2011) 072106.
[3] E. Pourali, M.K. Moravvej-Farshi, M. Ebnali-Heidan, Design and analysis of
slow light regime in silicon carbide 2D photonic crystal waveguides, Infrared
Phys. Technol. 63 (2014) 10e16.
[4] L. Meng, P. Meng, Q. Zhang, Y. Wang, Fast screening of ketamine in biological
samples based on molecularly imprinted photonic hydrogels, Anal. Chim. Acta
771 (2013) 86e94.
[5] J.P. Walker, K.W. Kimble, S.A. Asher, Photonic crystal sensor for organophos-
phate nerve agents utilizing the organophosphorus hydrolase enzyme, Anal.
Bioanal. Chem. 389 (2007) 2115e2124.
[6] K. Wrobel, D.M. Rodriguez, F.J.A. Aguilar, K. Wrobel, Determination of meth-
anol in o,o-dimethyldithiophosphoric acid (DMDTPA) of technical grade by
UV/vis spectrophotometry and by HPLC, Talanta 66 (2005) 125e129.
[7] J. Schlatter, F. Chiadmi, V. Gandon, P. Chariot, Simultaneous determination of
methanol, acetaldehyde, acetone, and ethanol in human blood by gas chro-
matography with flame ionization detection, Hum. Exp. Toxicol. 33 (2014)
74e80.
[8] W.S. Cheng, Z.C. Qi, B.Y. Chen, Determination of methanol in cosmetics by
sulfurous acid and basic fuchsine colorimetry, J. Environ. Health 21 (2004)
58e59.
[9] G.L. Wang, L.M. Gao, G.X. Ning, J.D. Cheng, Determination of methanol in
methylamine manufacturing wastewater by chromotropic acid spec-
trophotography, Admin. Techn. Environ. Monit. 4 (2008) 37e39.
[10] Y. Zhang, Q. Fu, J. Ge, Photonic sensing of organic solvents through geometric
study of dynamic reflection spectrum, Nat. Commun. 6 (2015) 7510e7516.
[11] C. Fenzl, T. Hirsch, O.S. Wolfbeis, Photonic crystals for chemical sensing and
biosensing, Ange, Chem. Int. Ed. 53 (2014) 3318e3335.
[12] O. Sato, S. Kubo, Z.-Z. Gu, Structural color films with lotus effects, super-
hydrophilicity, and tunable stop-bands, Acc. Chem. Res. 42 (2008) 1e10.
[13] K. Liu, T. Schmedake, R. Tsu, A comparative study of colloidal silica spheres:
photonic crystals versus Bragg's law, Phys. Lett. A 372 (2008) 4517e4520.
[14] F. Wang, Z. Zhu, M. Xue, F. Xue, Q. Wang, Z. Meng, W. Lu, W. Chen, F. Qi, Z. Yan,
Cellulose photonic crystal film sensor for alcohols, Sensors Actuat. B-Chem.
220 (2015) 222e226.
[15] J.Y. Shieh, J.Y. Kuo, H.P. Weng, H.H. Yu, Preparation and evaluation of the
bioinspired PS/PDMS photochromic films by the self-assembly dipedrawing
method, Langmuir 29 (2013) 667e672.
[16] B. Hatton, L. Mishchenko, S. Davis, K.H. Sandhage, J. Aizenberg, Assembly of
large-area, highly ordered, crack-free inverse opal films, Proc. Natl. Acad. Sci.
107 (2010) 10354e10359.
[17] S. Thomas, J. Guild, The CIE colorimetric standards and their use, Trans. Opt.
Soc. 33 (1931) 73e134.
[18] R.W.G. Hunt, M.R. Pointer, Measuring Colour, fourth ed., Wiley, Chichester,
2011.
[19] J. Li, T. Zheng, A comparison of chemical sensors based on the different or-
dered inverse opal films, Sens. Actuators B 131 (2008) 190e195.
[20] C. Fenzl, T. Hirsch, O.S. Wolfbeis, Photonic crystal based sensor for organic
solvents and for solvent-water mixtures, Sens 12 (2012) 16954e16963.