This article has been accepted for publication in IEEE Sensors Journal.

This is the author's version which has not been fully edited and
content may change prior to final publication. Citation information: DOI 10.1109/JSEN.2023.3305464

IEEE SENSORS JOURNAL, VOL. XX, NO. XX, MONTH X, XXXX 1

WaveFlex Biosensor: A Flexible-Shaped

Plasmonic Optical Fiber Sensor for Histamine
Detection
Wen Zhang, Ragini Singh, Feng-Zhen Liu, Carlos Marques, Bingyuan Zhang,
Santosh Kumar, Senior Member, IEEE

Abstract—In this work, a combination of

simulation and experiment was used to analyze the
distribution of the evanescent field in a W-shaped
optical fiber. A novel flexible W-shaped optical
fiber (FWOF) biosensor based on the localized
surface plasmon resonance effect (also known as
a WaveFlex biosensor) is designed and fabricated
for detecting histamine in food. First, gold
nanoparticles are immobilized on the FWOF
probe's surface to excite the LSPR effect. Second,
niobium carbide MXene and molybdenum disulfide
nanoparticles are immobilized on the sensing area
of the FWOF probe to improve the reaction area. A
larger surface area can offer more immobilization
sites for biomolecules. Meanwhile, to enhance the
specificity of the sensing probe, the diamine
oxidase enzyme is functionalized on the nanomaterials (NMs)-immobilized probe. The absorption spectrum and
microscopic distribution of the nanoparticles are characterized and measured using a UV-visible spectrophotometer
and a high-resolution transmission electron microscope. The immobilization results of NMs on the probe surface are
verified by scanning electron microscopy. In order to further explore the performance of the sensor, pH testing,
stability testing, reproducibility, repeatability, and probe specificity testing are also carried out. The proposed sensor
has a linear detection range of 0-1000 μM, a detection limit of 52.5 μM, and a sensitivity of 4.4 pm/μM.
Index Terms—WaveFlex biosensor; W-shaped optical fiber; optical fiber sensor; localized surface plasmon
resonance; histamine detection

I. Introduction vomiting, rash, diarrhea, and hypotension. Histamine poisoning

H
incidents occur frequently around the world, which not only
istamine is an amine substance produced by the cause the waste of food resources but also have a negative
decomposition of free histidine under the action of impact on consumer health. As a result, strict standards for the
microbial histidine decarboxylase [1]. It is a common limit of histamine in aquatic products are required. The Food
and most toxic biological amine that widely exists in organisms and Drug Administration stipulates that the histamine content
and a variety of aquatic products. When the single histamine of imported aquatic products not reach 450 μM. The European
intake of the human body reaches 50 mg, it will cause the Union requires that the histamine content of aquatic products
relaxation of the body's capillaries and the contraction of and their derivatives not surpass 0.9-1.8 mM [4]–[6]. Histamine
gastrointestinal smooth muscle, which will lead to histamine content, as one of the biomarkers of food quality, has attracted
poisoning [2], [3]. The clinical manifestations are nausea, much attention. Therefore, it is particularly critical to develop a

This work was supported by the Double-Hundred Talent Plan of of Physics Sciences and Information Technology, Liaocheng University,
Shandong Province, China; Liaocheng University (318052341), the Liaocheng 252059, China (e-mail: [email protected],
Special Construction Project Fund for Shandong Province Taishan [email protected], [email protected]).
Mountain Scholars; and the Natural Science Foundation of Shandong R. Singh is with the College of Agronomy, Liaocheng University,
Province (ZR2020QC061). This work was also developed within the Liaocheng 252059, China (e-mail: [email protected]).
scope of the projects i3N by the Fundacao para a Ciencia e a Tecnologia Feng-Zhen Liu is with the Liaocheng People’s Hospital, Medical
under grant 2021.00667.CEECIND; i3N projects LA/P/0037/2020, College of Liaocheng University, Liaocheng 252000, China (e-mail:
UIDB/50025/2020, and UIDP/50025/2020; and DigiAqua Project [email protected]).
PTDC/EEI-EEE/0415/2021. (Corresponding authors: Santosh Kumar Carlos Marques is with the Department of Physics, I3N, University of
and Bingyuan Zhang). Aveiro, 3810-193 Aveiro, Portugal (e-mail: [email protected]).
W. Zhang, B. Zhang and S. Kumar are with Shandong Key
Laboratory of Optical Communication Science and Technology , School

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8 IEEE SENSORS JOURNAL, VOL. XX, NO. XX, MONTH X, XXXX

sensor with fast detection speed, low cost, high accuracy, and of the noble MNPs, thereby realizing the quantitative detection
real-time detection of histamine. of the target analyte [18], [19]. In contrast to conventional fiber-
To date, a variety of methods have been used by researchers optic biosensors, LSPR-based biosensors would not require
to detect histamine. For example, Tian et al. [7] proposed a labeling of molecular recognition elements, are simple to
temperature-regulated biosensor of electroactive phase-change operate, and are beneficial to protecting the activity of
microcapsules (EPM) that innovatively combined EPM and biological cell molecules. As an emerging sensor, LSPR-based
diamine oxidase (DAO) to achieve temperature regulation of optical fiber biosensors have the advantages of high sensitivity,
the working electrode. It has practical application in the high resolution, fast response speed, and strong anti-
detection of histamine. Tian et al. [8] proposed a colorimetric interference ability, and they have a significant application in
histamine sensor in fish based on magnetic DAO-induced gold environmental monitoring, biomedicine, and biochemical
nanorod etching. The designed colorimetric biosensor shows a sensing.
good response to histamine, which has a certain application Nanomaterials (NMs) have novel planar spatial
prospect in the quality detection of aquatic products. configurations and unique physical and chemical properties,
Additionally, high performance liquid chromatography, that inject new impetus into the development of WaveFlex
chemical colorimetry, capillary electrophoresis, thin-layer biosensors [20]. The rational utilization of NMs can create
chromatography, and other techniques [6] have been better conditions for the immobilization of biometric materials,
extensively used to detect histamine. However, these detection which can improve the stability and immobilization of
methods have disadvantages such as high professional skill biological components, thereby significantly improving the
requirements, low sensitivity, and high instrument operating sensor's sensitivity and detection range. Gold nanoparticles
costs, making it difficult to meet the demands for convenient (AuNPs) occupy an increasingly important position in the field
and on-site detection. of LSPR sensing due to their superior optical qualities, easy
In recent years, fiber optic-based sensors have gained surface modification, good stability, and excellent
popularity among researchers, like side-polished [9], V-groove biocompatibility [21]. During the development of fiber optic
[10], and taper optical fiber (TOF) structures. Similarly, sensors, two-dimensional (2D) materials such as niobium
Esposito [11] investigated chemical sensors based on long- carbide (Nb2CTx) MXene, and molybdenum disulfide
period gratings (LPGs) and paved the way for the advancement nanoparticles (MoS2-NPs) are often used to improve their
of fiber optic sensors based on LPGs technology. In addition, sensing performance. The unique structure, composition, and
numerous grating-based fiber optic sensors are employed in the physical and chemical properties of the 2D nanomaterial
biosensing field, such as LPGs-based sensors for C-reactive Nb2CTx MXene make it another hot material in the field of 2D
protein in human serum [12] and TFBGs-based sensors for NMs after graphene [22]. The application range of Nb2CTx
human epidermal growth factor receptor-2 proteins [13]. MXene extends from the fields of mechanics, optics,
The optical fiber sensor based on the lossy mode resonance electronics, and energy storage to biomedicine, environmental
(LMR) principle has also gained the favor of researchers. For protection, etc. [23]–[26]. This is due to its excellent stability,
example, Chiavaioli et al. [14] designed an LMR-based fiber layered structure with a large area of contact, good
optic sensor for the detection of tau protein, a biomarker of biocompatibility, high conductivity, and abundant hydrophilic
Alzheimer's disease. Similarly, Moro et al. [15] proposed a high functional groups. Excellent hydrophilicity enables it to adsorb
performance and multifunctional sensing strategy based on more biomolecules and improve detection sensitivity. In the
LMR for the determination of monofluoroalkyl and MoS2-NPs structure, molybdenum atoms are connected with
polyfluoroalkyl substances in environmental samples. two layers of sulfur atoms by covalent bonds. This structure
Choudhary et al. [16] devised a fiber optic sensor based on makes MoS2-NPs more stable, not only as an effective bridge
LMR, which was experimentally demonstrated to have between enzymes or molecules and NPs, but also as an effective
potential for biochemical sensing applications. Additionally, it way to improve the loading capacity of the probe surface [27].
has low temperature cross-sensitivity. It has been widely used in research fields such as catalysis,
In recent years, the development of (localized) surface sensing, biology, drug loading, and treatment [28]–[31].
plasmon resonance (SPR/LSPR)-based sensors has received a In this work, a FWOF probe is fabricated using a
lot of attention from the research community. The optic-fiber conventional single-mode fiber (SMF). AuNPs, MoS2-NPs, and
LSPR sensor has attracted extensive attention from scientists Nb2CTx MXene are immobilized on the probe to improve the
because of its small size, low detection cost, high sensitivity, sensing performance, and DAO enzyme is used to improve the
and strong anti-interference ability. It has a wide range of specificity of the WaveFlex biosensor. The developed probes
applications in biomedicine and environmental monitoring and synthesized NMs are characterized before being tested on
[17]. Compared with SPR, LSPR has the characteristics of a the analyte. Finally, the sensing performance (selectivity,
short attenuation length of the electromagnetic field, less stability, reproducibility, pH, and reusability test) of the sensor
environmental impact, simple design, high flexibility, and high is evaluated and found to be satisfactory.
sensitivity to the surface biochemical interaction of metal
II. EXPERIMENTAL SECTION
nanoparticles (MNPs). The LSPR-based optical fiber biosensor
was designed and proposed by noble MNPs excited by the A. Materials and reagents
evanescent waves (EWs) generated by the fiber-optic
configuration. The interaction of the target analyte and the The FWOF structure was fabricated using traditional SMF (9
ligand on the sensor surface causes changes in the surrounding μm, 125 μm, Shenzhen Technologies Co.,Ltd). Acetone was a
RI, which results in a shift in the absorption peak wavelength ketone cleaning agent that was used to remove various organic

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8 IEEE SENSORS JOURNAL, VOL. XX, NO. XX, MONTH X, XXXX

compounds from the surface of the probe. Tetrachloroauric acid noble MNPs and target capture substances
(HAuCl4), trisodium citrate (C6H5Na3O7) and deionized (DI) (enzymes/antibodies) on the surface of the probe by covalent
water were mainly used to synthesis the AuNPs. In the process bonding. When light passes through the sensing area, the NMs
of attaching enzyme to the probe surface, necessary reagents absorb light to generate plasmon resonance, and the detector
such as 11-mercaptoundecanoic acid (MUA), n- collects the plasmon resonance spectrum. When the capture
hydroxysuccinimide (NHS), and N-(3-dimethylaminopropyl)- elements immobilized on the NMs are specifically combined
n-ethylenediimide hydrochloride (EDC) were used. MoS2 with the analyte, the RI around the NMs immobilized on the
powder was purchased from Sigma-Aldrich and for the surface of the fiber changes, which in turn causes the position
preparation of MoS2-NPs, and N-N-methyl-2-pyrrolidone of the resonant peak in the LSPR characteristic spectrum to shift
(NMP) was used as a solvent to dissolve MoS2 nanosheets. [33]. The resonance peak shift and the RI of the medium
Nb2CTx Mxene was purchased from Xianfeng Nano Material surrounding the MNPs have the following relationship [34]:
Technology Co., Ltd., China and N,n-Dimethylformamide "#$
(DMF) solution was used to dissolve Nb2CTx Mxene. ∆𝜆 = 𝑚∆𝑛! &1 − 𝑒 %! * (1)
B. Measurement and instrument
Among them, 𝑚 is the sensitivity factor of the NPs, ∆𝑛!
The FWOF structure was fabricated using the powerful is the change in the RI of the NPs surface medium, 𝑙 is the
processing instrument of the combiner manufacturing system effective thickness of the bound substance on the NPs surface,
(CMS, USA). To determine the size of the NPs, the ultraviolet- and 𝑑& represents the attenuation length of the electromagnetic
visible (UV-Vis) spectrophotometer (Hitachi, Japan) was used
field. According to Eq. (1), the performance of LSPR
to measure the absorption spectra of AuNPs and MoSo2-NPs.
biosensors can be analyzed by observing a change in RI when
The microscopic distribution of NPs synthesized in the
a substance binds to the surface of NPs. This conversion of a
experiment was characterized using high-resolution
small RI change near the metal surface into a detectable
transmission electron microscopy (HR-TEM, Thermo Fisher
wavelength shift response is crucial for detecting and
Scientific, USA). To observe the coating of NMs on the surface
measuring substances using LSPR biosensors. This is the
of the sensing probe, a scanning electron microscope (SEM,
theoretical basis for realizing the WaveFlex biosensor.
Carl Zeiss Microscopy, Germany) was used to scan the sample
The formation of effective evanescent fields is critical to
by emitting a beam of high-energy electrons. The tungsten-
the performance of WaveFlex biosensors. EWs can enhance the
halogen light source (Ocean Optics, USA) can emit a wide
electric field in a certain range of penetration depth, which is
spectrum. A spectrometer (Ocean Optics, USA) was used to
beneficial to the LSPR phenomenon of noble MNPs [35]. In
collect experimental data and investigate the light transmission
traditional SMF, the core has a higher RI than the cladding,
characteristics of the WaveFlex biosensor.
causing the beam to be confined and propagate within the core.
C. Sensing principle The energy carried by the higher-order modes is leaked into the
cladding in the form of EWs, but the energy carried by the
LSPR is a nano-optical technology, that is based on the higher-order modes is very small, so it is difficult to form a
interaction between light and noble MNPs on a scale smaller strong evanescent field. The EWs energy generated by the weak
than the incident light wavelength. When incident light's evanescent field will be lower, so the phenomenon is not
electric field interacts with free electrons in NPs, charge obvious when the SPR is excited. Compared with SMF, the
separation between the free electrons and the metal nucleus binding ability of TOF to light is much lower. The mode of
occurs. Coulomb repulsion between free electrons causes them transmission light in TOF changes, and the energy of
to move in opposite directions, resulting in electron collective transmission light will be coupled into the cladding to form a
oscillation [32]. The plasmon resonance of MNPs is shown in strong evanescent field [18].
Fig. 1(a). LSPR-based sensors usually need to immobilize
(a)

(b)

Fig. 1. Schematic of (a) LSPR phenomena, (b) wave propagation through proposed WaveFlex sensor probe.

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In addition, the TOF structure is simple to fabricate, cost- E. Fabrication of sensing probe
effective, and highly sensitive, with high absorbance sensitivity
In this work, the fiber taper processing was achieved by heating
for EWs. The EWs that leak in the tapered region interact with
the surrounding medium, which changes the transmission the fiber to a molten state by the arc discharge method and
gradually stretching it under the action of tension. The CMS
mode, spectrum, light intensity, and other characteristics of the
fiber. It can be observed through the transmission spectrum to provided the most powerful tool for supporting the design and
achieve the purpose of sensing. The tapered region of FWOF is fabrication of novel FWOF structure. The proposed FWOF
more suitable for generating high-power EWs. Compared with structure was processed using a three-electrode high thermal
TOF, FWOF has a larger surface area, which is beneficial for stability plasma heating system, an innovative technology
developed for the CMS instrument. CMS worked in a partial
increasing the contact space between the EWs and the external
environment, thereby enhancing the sensor's sensitivity. The vacuum environment, which increased the long-term discharge
FWOF sensor probe's schematic is shown in Fig. 1(b). stability and made it easy to obtain target structures with good
repeatability and quality. The CMS machine is a semi-
D. Simulation of Sensor Probe automatic fiber processing system with a wider range of
position adjustment and sufficient capacity for fiber taper. After
In order to explore the transmission state of light inside the
setting the parameters of the program according to the needs of
FWOF probe in detail, the beam propagation method (BPM) in
the target structure, the CMS can automatically execute the
RSoft software was used to simulate the designed fiber
tapering process under the control of the program. The
structure, and the energy distribution of light propagating in the
important parameters of the program included the specific
sensing area was determined, as depicted in Figs. 2 (a) and (b).
length and diameter of the taper, vacuum value, electrode
The BPM can analyze the optical path and monitor specified by
discharge power, and motor moving speed. The internals of the
the user and study the optical properties of each part of the fiber
CMS machine are shown in Fig. 3(a). For consideration of
structure in real time. Fig. 2(b) illustrates the energy distribution
mechanical strength and sensing performance, procedures with
of the evanescent wave when the light propagates in the tapered
waist diameters of 40 μm and 60 μm were chosen to design the
fiber. The 1, Launch was the total energy distribution of the
probe. Under the program of CMS, bidirectional tapering, the
light source incident into the fiber core. And the 1, Mode 0 and
tapering process and parameters of each part of the FWOF
2, Mode 0 represented the energy distribution of the
probe are depicted in Fig. 3(b).
propagation mode in the core and cladding, respectively. When
the light is transmitted into the tapered region, the energy of the F. AuNPs/ Nb2CTx Mxene/MoS2-NPs synthesis process
core mode drops sharply, while the energy of the cladding mode
By using the conventional Turkevich method [36], spherical
increases sharply. Part of the light coupled with the
fundamental mode continues to propagate in the fiber core, AuNPs with a particle size of approximately 10 nm were
while part of the light leaks into the outer medium to form EWs. synthesized. MoS2-NPs were synthesized by the liquid-phase
The final total output power value of the light is 0.9, which is ultrasonic method [28]. Firstly, 10 mL of NMP solution was
used to dissolve 30 mg of MoS2 powder, followed by 1 hour in
mainly due to the loss of energy generated by the EWs formed
in the tapered region, and the loss was determined by the an ultrasonic water bath. Secondly, the solution after heating in
tapered structure and the external medium. The simulation the water bath was treated for 10 minutes in an ultrasonic cell
results verified that more EWs are excited at the tapered waist crusher with a power of 1000 W, with the aim of breaking large-
size MoS2-NPs into small-size. Finally, the obtained solution
region of the FWOF structure, which is conducive to the LSPR
effect with external materials to achieve the purpose of sensing. was centrifuged at 4000 rpm for 1 hour, and the transparent
upper layer solution was collected for further use. To make the
(a) (b) Nb2CTx MXene [19] dispersion, 2 mg of Nb2CTx Mxene was
first dissolved in 10 mL of DMF. Subsequently, the mixed
solution was treated with an ultrasonic machine for 1 hour to
obtain the dispersion of Nb2CTx Mxene.
G. Nanomaterials immobilization and enzyme
functionalization
It was necessary to perform chemical molecular modification
on the sensing region of the sensor in order to carry out the
sensing function. The surface chemistry properties determined
the stability, specificity, and reproducibility of the sensor. The
specific steps of the modification scheme were as follows: First,
clean the sensing area of the probe with an acetone solution for
20 minutes to remove impurities from the probe surface. The
probe was then immersed in Piranha solution (mixture of 3
volumes of 30% H2O2 and 7 volumes of concentrated H2SO4)
Fig. 2. (a) Light propagation in FWOF probe, (b) plot of energy propagation for 30 min. After treating the probe surface, the surface was
mode inside the FWOF probe. hydroxylated, and the organic matter was washed away to
immobilize AuNPs on the probe. Next, probe was immersed in
the 1% MPTMS ethanol solution for 12 hours. The hydroxyl

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8 IEEE SENSORS JOURNAL, VOL. XX, NO. XX, MONTH X, XXXX

20 minutes. The dip-coating process was carried out three times

to improve the stability of Nb2CTx MXene immobilization.
Afterwards, the probe was immersed in 10 mL of MoS2-NPs
solution for 30 s and dried at 50 °C for 2 minutes. This process
was carried out eight times to make sure that the MoS2-NPs
layer uniformly covered the surface of the fiber optic probe, and
then the probe was heated in the oven at 60 °C for 2 hours to
further immobilize the NMs.
In order to realize the specific detection of histamine, it
was necessary to modify the fiber with a layer of enzyme that
can specifically bind to histamine on the sensor's surface. The
covalent binding method not only made the enzyme and carrier
have higher binding efficiency, but also had better enzyme
stability. Therefore, the enzyme was immobilized on the probe
by covalent bonding, and the enzyme functionalization steps of
the probe were as follows: The probe was immersed in 10 mL
of a 0.5 mM MUA ethanol solution for 5 hours to modify the
surface with carboxyl groups. Subsequently, the sensing area
was dipped in 5 mL of a mixed solution of EDC (200 mM) and
NHS (50 mM) for 30 minutes to fully activate the carboxyl
group. EDC is a non-toxic and biocompatible cross-linking
reagent that causes the amino groups of the DAO to cross-link
with the carboxyl groups of the sensor surface. After 30
minutes, the probe was washed with DI water and immediately
soaked in a freshly made DAO enzymatic solution for 6 hours,
and the amino group of the enzyme formed a covalent bond
Fig. 3. (a) Internal layout of the CMS machine, (b) fabrication procedure of
with the activated carboxyl group to realize the
FWOF structure. functionalization of the enzyme. The process of NMs
immobilization and DAO functionalization on the FWOF
groups on the surface of the probe and MPTMS undergo a surface is shown in Fig. 4.
hydrolysis reaction to form Si-O-Si bonds, thereby forming
self-assembled groups with amino groups on the probe's H. Preparation of histamine samples
surface. The treated probes were washed with deionized water Different concentrations of histamine solution were prepared in
and dried in the oven at 70°C for 30 min. Then, the MPTMS- the concentration range of 0-1000 μM. Accurately weigh 5.56
coated probe was placed in a freshly synthesized AuNPs
mg of histamine powder, dissolve it in 1 × PBS (pH=7.4)
solution for 48 hours to realize the self-assembly of AuNPs on solution, and make it into 50 mL to obtain a concentration of
the probe sensing region. The probe was rinsed with deionized
1000 µM base solution. Dilute the stock solution with 1×PBS,
water and dried with N2 gas to remove unbound AuNPs from
and prepare eight histamine solutions with different
the surface. Next, the probe was immersed in the Nb2CTx
concentration gradients (0, 10, 50, 100, 200, 400, 600, and 800
MXene solution for 10 min, and dried in the oven at 70 °C for
μM) for testing separately.

Fig. 4. Nanomaterials-immobilization and DAO enzyme-functionalization over the WaveFlex sensor probe.

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8 IEEE SENSORS JOURNAL, VOL. XX, NO. XX, MONTH X, XXXX

(a) (b)

330 nm
519 nm

(c) (d) (e)

100 nm

Fig. 6. (a) AuNPs absorbance, (b) MoS2-NPs absorbance, HR-TEM images of

(c) AuNPs, (d) MoS2-NPs, and (e) Nb2CTx MXene.
Fig. 5. Schematic of histamine reaction process and experimental setup for
histamine detection. B. Optimization of sensor probe
I. Experimental setup CMS is the main tool for fabricating FWOF probes. Compared
with the traditional hydrogen-oxygen flame heating method, the
The fusion splicer machine (FSM) was used to connect the method of using CMS machine has the advantages of high
probe with the source and spectrometer. The tungsten-halogen efficiency, high repeatability, and high stability. CMS arc
lamp light source was used to excite the LSPR effect of AuNPs technology provides a controlled plasma field for uniform heat
on the probe's surface. Histamine was specifically bound to the distribution, supported by a three-electrode heating mode. The
DAO on the probe surface, and the decomposed byproducts three-electrode arc surrounds the uniformly fused SMF during
attach to the surface of AuNPs, resulting in changes in the RI the discharge process and is gradually stretched under the action
around them. Fig. 5 depicts the optical measurement setup for of the platform’s moving tension to form a tapered structure.
detecting various concentrations of histamine. The permittivity The TOF's specific size parameters can be preset in the
of the AuNPs also changed, which in turn created new program, and the parameters are continuously optimized and
resonance frequencies. The LSPR spectrum's resonant peak was debugged until the desired goal is achieved. Fig. 7(a) is a
red-shifted at this moment, and the concentration of the diagram of the FWOF diameter structure obtained by the CMS
histamine can be further determined through the wavelength scanning function. It can be seen from the results that the
shift, thereby realizing the detection of the histamine with the diameter of the taper waist is very uniform, and the transition is
schematic shown in Fig. 5. smooth, reflecting the stability of the parameters of the taper
procedure. Figure 7(b) depicts the transmission spectra of the
III. RESULTS AND DISCUSSIONS fabricated WaveFlex sensors. According to the results, the
A. Characterization of nanomaterials initial peak positions of the spectra of each sensor are almost
identical.
The characterization of NMs is a key aspect of the
development of WaveFlex biosensors. The synthesized AuNPs C. Characterization of nanomaterials-immobilized
solution is characterized using a UV-visible spectrophotometer. structure
There is an obvious absorbance peak at 519 nm, as shown in The surface topography of the sensor affects the modification
Fig. 6(a), that confirms that AuNPs with a diameter of and distribution of NPs on the sensor. Fig. 8(a) shows the SEM
approximately 10 nm were synthesized. The particle size and results of the WaveFlex biosensor, which suggest that the
dispersion of AuNPs also determine the efficiency and effect of sensing probe modified with NMs is a good foundation for the
AuNPs modification on the sensor surface. Also, Fig. 6(b) subsequent measurement experiments. Furthermore, the
illustrates the MoS2-NPs absorption spectrum, with an number and degree of dispersion of AuNPs on the sensor affect
absorption peak wavelength of 320 nm, which proves that the signal strength of the sensor. The AuNPs on the sensor are
relatively stable MoS2-NPs are synthesized. The distribution densely distributed and evenly dispersed, indicating that the
and morphology of AuNPs in the solution are captured by HR-
TEM and shown in Fig. 6(c). The synthesized NPs have good
spherical dispersion and consistent particle size. The uniform
size is conducive to the excitation of LSPR. The TEM image of
MoS2-NPs is shown in Fig. 6(d), and the results demonstrate
that small-size (2-3 nm) MoS2-NPs are successfully
synthesized. Nb2CTx MXene is also characterized using HR-
TEM, the geometry of Nb2CTx MXene is a layered structure,
as shown in Fig. 6(e), which is advantageous for increasing
DAO binding sites. Fig. 7. (a) Diameter analysis, and (b) transmitted intensity of FWOF probe.

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0 µM
1.0 (a)

Normalized intensity (a.u.)

10 µM
20 µM
50 µM
100 µM
0.8 200 µM
400 µM
600 µM
0.6 800 µM
1000 µM

0.4

0.2

660 662 664 666 668 670 672

400 500 600 700 800 900

Wavelength (nm)

667 (b)

Peak wavelength (nm)

666 方程 y = a + b* x
绘图 Pe a k wa vel e ngt h

Fig. 8. (a) SEM image of FWOF probe surface, (b) SEM image of NPs- 截距 661. 96009 ± 0. 03945
斜率 0. 00426 ± 1. 71965E- 4

immobilized probe (c) EDX of AuNPs/ MoS2-NPs/ Nb2CTx MXene- 665

immobilized probe.
664
AuNPs are successfully immobilized on the FWOF probe's λ = 0.0044! + 661.94
surface. The SEM results of AuNPs, MoS2-NPs and Nb2CTx 663 R² = 0.9857
MXene immobilized on the surface of the structure are also
clearly shown in Fig. 8(b). The elemental composition of the 662
immobilized NMs is verified by SEM-EDX analysis. The result
661
is shown in Fig. 8(c). The presence of Au, Mo, and Nb elements 0 200 400 600 800 1000
confirmed that the fiber surface is successfully coated with
Concentration of Histamine (µM)
AuNPs, MoS2-NPs, and Nb2CTx MXene, respectively.
Fig. 9. (a) LSPR sensing spectrum of histamine solutions, (b) linearity result
D. Measurement of analytes of WaveFlex biosensor.
The sensing ability of the sensor is evaluated by recording the Sensitivity is an essential criterion for measuring sensor
change of the sensor probe in response to histamine performance. The higher the sensor's sensitivity, the greater the
concentrations ranging from 0 μM to 1000 μM. The spectral response to small signal changes, and the higher the possibility
changes of three different sensing probes are tested in sequence of application in actual biomedicine. It can be concluded that
while the external environment remains constant. The the probe's sensitivity is 4.4 pm/µM, and the correlation
normalized transmitted intensity spectra of the three sensing coefficient is 0.98. It shows that the probe has good linearity. In
probes are averaged, and shown in Fig. 9(a). The observed the actual biosensor detection, a more important indicator is
phenomenon indicates that an increase in the concentration of involved: the limit of detection (LoD). We define the LoD as
the histamine solution leads to a corresponding shift of the peak the smallest detectable signal caused by analyte interaction,
wavelength towards longer wavelengths. Furthermore, it is calculated as:
noteworthy that the magnitude of this shift is directly
proportional to the concentration of histamine, with higher 3 × 𝑆𝐷
𝐿𝑜𝐷 = (3)
concentrations resulting in greater shifts. This is due to the 𝑆𝑒𝑛𝑠𝑖𝑡𝑖𝑣𝑖𝑡𝑦
catalytic action of the DAO enzyme on histamine. The The LoD tested according to Eq. 3 is 52.5 µM. WaveFlex
generation of by-products leads to a change in the RI of the biosensors, as the name implies, are biosensors developed on
AuNPs on the sensor, which shifts the peak position of the the basis of plasmonic waves that are sufficiently flexible and
plasmon resonance peak and is detected by the spectrometer. supple. This work results in the development of an innovative
Since the peak position shift correlates positively with WaveFlex biosensor that is experimentally verified and suitable
histamine concentrations, the quantitative detection of
for diverse applications, such as environmental monitoring and
histamine in the sample is realized by detecting the shift of the
food safety detection.
plasma resonance peak.
The linear fitting equation for the peak wavelength E. Stability and pH test
changing with the histamine solution is shown in Fig. 9(b). The
A stability test is usually employed to investigate the response
fitting equation is:
of the sensor's performance across a range of measurements.
𝜆 = 0.0044 𝐶 + 661.94 (2)
Through continuous measurements, it is determined whether
Here, 𝐶 (μM) is the concentration of the histamine sample, 𝜆 the performance of the sensor remains stable over time. The
(nm) represents the peak wavelength of the LSPR spectrum. sensing probe is placed in PBS, and the spectral data of the
sensor is recorded periodically after the sensor has completed

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(a) (b)

Fig. 10. (a) Stability, and (b) pH test of proposed probe.

its response. Figure 10(a) shows the fluctuation of the initial

peak wavelength of the probe during fifteen measurements. The
experimental standard deviation (SD) can be obtained as 0.077.
It is shown that NPs and enzyme coatings are stable on the Fig. 12. Selectivity test of sensor probe.
sensor surface, which verifies the good stability of the designed
biosensor. The purpose of pH testing is to determine the pH G. Selectivity test
value most favorable to the solvent environment of the analyte. In addition to studying the probe's sensitivity and detection
Prepare histamine sample solutions with concentrations of 0 range, the selectivity of the sensor is also a very important
µM and 1000 µM in a solvent environment with pH=3, 6, 7.4, indicator. Selectivity refers to the capacity of the sensor to
10, and 13. Test from low to high pH values, and at the same detect the measured substance in a complex environment, and
pH value, measure the low concentration (0 µM) first, then the the selectivity of the sensor determines whether the sensor has
high concentration (1000 µM). Each time, an equal amount of the possibility of application in practice. The sensor's response
the liquid to be tested is injected into the reaction tube, and the to six different interferents (Spermine, Tyramine, Putrescine,
corresponding LSPR spectrum is recorded. The results are Tryptamine, L-Cysteine, and Sarcosine) is investigated. Low-
shown in Fig. 10(b), the abscissa represents the liquid concentration (0 μM) and high-concentration (1000 μM)
environment with different pH, and the ordinate is the biomolecule solutions are prepared in PBS for testing, and the
resonance wavelength shift between the corresponding differences in resonance wavelength peaks from low-
concentrations. The results show that the maximum drift occurs concentration solutions to high-concentration solutions are
in a liquid environment with a pH of 7.4, which validates the analyzed. Fig. 12 shows the shift of the sensor's resonance
rationality of selecting PBS at pH=7.4 as the sample buffer in wavelength in all interfering substance solutions, and the shift
this experiment. of the LSPR peak of the sensor for histamine is much higher
F. Reproducibility and Reusability test than that for other substances, demonstrating the sensor’s
excellent specificity recognition ability.
Reproducibility refers to the capacity to evaluate the same
concentration of histamine with the same batch of sensing H. Evaluation of sensor performance
instruments and obtain identical results. At 1000 μM of Table 1 compares the detection range, LoD, and sensitivity
histamine, the reproducibility of the sensing probes is assessed. between the fiber optic LSPR sensor constructed in this
The recorded spectra are shown in Fig. 11(a), and the three sets experiment and other common detection methods for histamine.
of waveforms are generally highly consistent, which proves that Although some detection methods are superior to the optical
the developed probe has good reproducibility. fiber sensor proposed in this work in terms of LoD, these -
Reusability is an important indicator for assessing the
TABLE 1. PERFORMANCE COMPARISON OF THE PROPOSED
price and effectiveness of probes. The sensor probe's ability to SENSOR WITH THAT OF EXISTING HISTAMINE SENSORS.
be reused was tested under different histamine samples (100
Materials used Mechanisms Linear Sensitivity LoD Ref.
μM and 800 μM). The normalized transmission spectrum range
results are depicted in Fig. 11(b), indicating that the probe has Magnetic graphene Colorimetric 5–160 µM n.r.a 1.23 [8]
good reusability. In addition to reusability, sensor regeneration oxide µM
should also be investigated. The use of regenerative solutions AgNPs Surface- 10− 8–10− 3 n.r.a 3.088 [38]
enhanced Raman M/L nM
to eliminate immobilized nanomaterials and bioreceptors is scattering
called regeneration [37]. Tungsten trioxide- Electrochemical 1 nM–1 n.r.a 0.17 [39]
(a) (b) NPs mM nM
AuNPs Colorimetric 0.001–10 n.r.a 0.87 [40]
µM nM
Carbon quantum Fluorescence 0.1–100 n.r.a 13 ppb [41]
dots ppm
Molecularly SPR 8.68 fM– n.r.a 5.04 [42]
imprinted 86.8 pM fM
polymers NPs
AuNPs/MoS2-NPs/ LSPR 0–1000 4.4 pm/µM 52.5 This
Nb2CTx MXene μM μM work
a
Fig. 11. (a) Reproducibility, and (b) reusability test of proposed probe. not reported

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8 IEEE SENSORS JOURNAL, VOL. XX, NO. XX, MONTH X, XXXX

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