T.R.

NECMETTIN ERBAKAN UNIVERSİTY

INSTITUTE OF SCIENCE

PAPER BASED OPTİCAL SENSORS

Manel AZLOUK

PhD SUBJECT PRESENTATION

Department Of Chemistry

SUPERVISOR
Prof. Dr. Haluk BINGOL

June-2018
KONYA
All Rights Reserved
ABSTRACT
The recent advances and state of the art in paper-based analytical devices (PADs) are shown
through the analysis of their integration with microfluidics and LOC micro- and nanotechnologies,
electrochemical/optical detection and electronic devices as the convergence of various knowledge areas.
The important role of the paper design/architecture in the improvement of the performance of sensor
devices is discussed. The discussion is fundamentally based on μPADs as the new generation of paper-
based (bio) sensors.
The main advantages and disadvantages of the use of paper as a substrate for sensors, as well as
its impact on their performance and application are presented,presents a short history of paper in
analytical chemistry, and discusses fabrication methods and available sources of paper.
Data about the scientific publication ranking of PADs, illustrating their increase as an
experimentalresearch topic in the past years, are supplied. In addition, an analysis of the simultaneous
evolution of PADs in academic lab research and industrial commercialization highlighting the parallelism
of the technological transfer from academia to industry is displayed. A general overview of the market
behaviour, the leading industries in the sector and their commercialized devices is given. Finally, personal
opinions of the authors about future perspectives and tendencies in the design and fabrication technology
of PADs are disclosed.

Keywords: analytical, device, diagnostics, lab-on-a-chip, microfluidics,paper, sensor .

 SEMİNER BİLDİRİMİ

Bu seminerdeki bütün bilgilerin etik davranış ve akademik kurallar çerçevesinde

elde edildiğini ve tez yazım kurallarına uygun olarak hazırlanan bu çalışmada bana ait
olmayan her türlü ifade ve bilginin kaynağına eksiksiz atıf yapıldığını bildiririm.

DECLARATION PAGE

I hereby declare that all information in this seminar document has been obtained
and presented in accordance with academic rules and ethical conduct. I also declare that,
as required by these rules and conduct, I have fully cited and referenced all material and
results that are not original to this work.

Manel AZLOUK

Date: 02.07.2018
 ACKNOWLEDGEMENT

I would like to express my gratitude to my supervisor Prof. Haluk BINGOL, for

the excellent supervision, help, guidance and encouragement he provided. In addition, I
am grateful to all the members of the GNM Research Group for their valuable help
during all phases of this work.

Most importantly, I would like to thank my family for their support,

understanding and love all these years.

Manel AZLOUK
KONYA-2018
 CONTENT

ABSTRACT................................................................................................................... iii

ACKNOWLEDGEMENT.............................................................................................iv

CONTENT.......................................................................................................................v

1. INTRODUCTION.......................................................................................................1

1.1. Chemical sensors....................................................................................................1

1.2. Sensor types............................................................................................................1

2. OPTICAL DETECTION............................................................................................2

3. FABRICATION AND PATERNING........................................................................3

3.1. Inkjet printing.........................................................................................................3

3.1. Photolithography....................................................................................................3

4. TRENDS OF PAPER BASED OPTICAL SENSORS.............................................3

3. MARKET OF PAPER BASED SENSORS...............................................................3

REFRENCES...................................................................................................................5

Curriculum vitae................................................................................................................8

v
 1

1. INTRODUCTION

1.1. Chemical sensors

The ideal chemical sensor is an inexpensive, portable, foolproof device that

responds with perfect and instantaneous selectivity to a particular target chemical
substance (analyte) present in any desired medium in order to produce a measurable
signal output at any required analyte concentration.
The committee defined chemical sensors as devices or instruments that
determine the detectable presence, concentration, or quantity of a given analyte. The
complexity of a chemical sensor application is related to the technical difficulties
associated with these determinations and with the specific nature (i.e., elemental or
molecular) of the chemical substance to be analyzed. Given the huge number (>10 6) of
known molecular substances, molecular sensing typically relies on recognition of
molecular structure or associated reactivity; this recognition aspect is called selectivity.
Sensitivity and limit of detection relate to the quantity or concentration of the element
or molecule to be analyzed (the analyte). The quantity of analyte present in a sample can
have a dynamic range of greater than 10 23, and chemical sensors are commonly required
to detect 10-9 molar concentrations or less. Thus, the challenge of attaining the needed
sensitivity in chemical sensing is comparable to that of achieving the needed selectivity.
The sensitivity and selectivity aspects of chemical sensing are affected by the
phase, dimensional, and temporal aspects of the desired determination. The analyte can
be present in a gas, liquid, or solid phase on various dimensional scales ranging from
bulk volumes of liters to picoliters, or surface layers from nanoscopic to monomolecular
scale. It may also be persistent or transitory. A further set of requirements can originate
from a need for repetitive measurements of the analyte over long times (e.g., days,
months) or at multiple and perhaps remote locations, such as in environmental analysis
and personal monitoring (Zor. E., 2012).
 2

Figure 1.1. Main principle of a chemical sensor

The design of chemical sensors also requires appreciation of the needed degree
of quantitative reliability (precision or accuracy). Finally, economic resources and
constraints can affect the design and strategy of any sensing task in many different
ways.
The capability of chemical sensing technology is substantial and has grown
steadily over the past several decades, but it has been outpaced by the needs and
diversity of chemical measurements. Materials limitations are prominent among the
existing limitations of chemical sensors. The following discussion outlines the various
chemical sensor types and strategies for further development, with emphasis on those
areas in which materials needs are especially evident (Singh et al., 2017)..

1.2. Sensor types

The development of instrumentation, microelectronics and computers makes it

possible to design sensors utilizing most of the known chemical, physical and biological
principles that have been used in chemistry. Chemical sensors may be classified
according to the operating principle of the transducer.
 3

1. Optical devices transform changes of optical phenomena, which are the result of
an interaction of the analyte with the receptor part. This group may be further
subdivided according to the type of optical properties which have been applied
in chemical sensors:
a) absorbance, measured in a transparent medium, caused by the absorptivity of
the analyte itself or by a reaction with some suitable indicator.
b) reflectance is measured in non-transparent media, usually using an
immobilized indicator.
c) luminescence, based on the measurement of the intensity of light emitted by a
chemical reaction in the receptor system.
d) fluorescence, measured as the positive emission effect caused by irradiation.
Also, selective quenching of fluorescence may be the basis of such devices.
e) refractive index, measured as the result of a change in solution composition.
This may include also a surface plasmon resonance effect.
f) optothermal effect, based on a measurement of the thermal effect caused by
light absorption.
g) light scattering, based on effects caused by particles of definite size present in
the sample.
The application of many of these phenomena in sensors became possible
because of the use of optical fibres in various configurations. Such devices have
also been called optodes. It should be emphasized that fibre optics now
commonly used are only technical devices applicable in a large group of optical
sensors which can be based on various principles (Chan et al., 2016).
2. Electrochemical devices transform the effect of the electrochemical interaction
analyte - electrode into a useful signal. Such effects may be stimulated
electrically or may result in a spontaneous interaction at the zero-current
condition. The following subgroups may be distinguished:
a) voltammetric sensors, including amperometric devices, in which current is
measured in the d.c. or a.c. mode. This subgroup may include sensors based on
chemically inert electrodes, chemically active electrodes and modified
electrodes. In this group are included sensors with and without (galvanic
sensors) external current source.
 4

b) potentiometric sensors, in which the potential of the indicator electrode (ion-

selective electrode, redox electrode, metaVmeta1 oxide electrode) is measured
against a reference electrode.
c) chemically sensitized field effect transistor (CHEMFET) in which the effect
of the interaction between the analyte and the active coating is transformed into
a change of the source-drain current. The interactions between the analyte and
the coating are, from the chemical point of view, similar to those found in
potentiometric ion-selective sensors.
d) potentiometric solid electrolyte gas sensors, differing from class 2b) because
they work in high temperature solid electrolytes and are usually applied for gas
sensing measurements.
3. Thermometric devices based on the measurement of the heat effects of a specific
chemical reaction or adsorption which involve the analyte. In this group the heat
effects may be measured in various ways, for example in the so called catalytic
sensors the heat of a combustion reaction or an enzymatic reaction is measured
by use of a thermistor. The devices based on measuring optothermal effects (If)
can alternatively be included in this group.
4. Other physical properties as for example X-, p- or r- radiation may form the
basis for a chemical sensor in case they are used for determination of chemical
compostion (Tunç, S., 2017).
 5

2. OPTICAL DETECTION

Detection by optical means is the most inexpensive and simplest method and is
universal . Nevertheless some variables have to be taken into account, such as the fact
that differences in illumination will cause variations in color intensity and hue. This
kind of problem can be solved by a white balance correction, subtraction of the
background color, or comparison with a calibration curve of standards of known color
and intensity. The latter solution can also be applied when different imaging devices are
used, e.g., when we want to compare results recorded by a camera, a scanner, and a
portable spectrometer. Several groups noted that quantification from hydrophobic paper
gave much better results than that from hydrophilic materials.
Droplets of reagents spread only a little on hydrophobic paper, resulting in
smaller, more densely packed detection regions, increasing the sensitivity of the assay
and facilitating quantification. It was also noted that hydrophilic paper could lead to
premature ending of the assay due to surface drying. The substrate could also entrap
sensing molecules too deeply, restricting accessibility. Irradiation with polychromatic
UV light before modification of paper could decompose phosphorescent impurities
found in Whatman 1PS filter paper, minimizing the phosphorescence background.
Another group noted that drying of the reagents at room temperature gave higher
background signals in comparison with papers dried in an incubator at 37 °C.
Uniformity of signal development is also a problem. In some cases reporting molecules
are carried out with liquid accumulating on the borders of the detection zone. In those
cases several simple pretreatment techniques can be applied to at least partially
immobilize or stabilize color development, e.g., with poly(vinyl amine), gelatin,
poly(acrylic acid), and poly(ethylene glycol).

2.1.1. Detectors and image analysis

A variety of detectors can be applied for optical sensing, from the commonest
devices such as scanners or camera phones to more specialized devices, including
spectrophotometers and fluorimeters, to sophisticated devices, e.g., gel documentation
systems. The simplest and least expensive method that does not require any additional
equipment is naked eye detection. As it can be influenced by different visual perception
of people or the aforementioned lightning conditions, this method should be used with
 6

color charts, which could be integrated on the devices. Differences between the color of
the reaction zone a dry printed paper and a wetted paper should be taken into account.
Another common device is an office scanner, which provides high resolution
and ensures focus of the digitalized image; furthermore, image intensity is not affected
by the external lighting conditions. Scanners can be portable (e.g., pen scanners), can
used by unskilled personnel, and are widely used all over the world. A digital camera
can also be used as a detector with automatic white balance, and also does not require
any special skills to use it. One shortcoming is that it cannot focus on objects that are
too close (approximately 20 cm). Cell phone cameras are also applicable, and it is worth
remembering that around 67 % of the world’s population owns a cell phone, with the
largest growth of ownership in the developing world. Userfriendly software could be
developed to unambiguously present results on the screen or the image could be sent to
a specialist for further analysis (telemedicine). More sophisticated cell-phone-based
readers are being developed, using the embedded flash of the cell-phone camera as the
light source, and piping it with an optical fiber exactly to the detection zone. They could
also provide pulses of energy required, for example, in electrochemiluminescence
sensing (Turner et al., 1987).
After digitalization, the image can be analyzed with the use of a computer
program such as Adobe Illustrator, Adobe Photoshop, GenePix, DigitalColor Meter,
Corel PhotoPaint, or ImageJ or other personalized software written in Python,
MATLAB, etc. Images can be converted to color spaces: CMYK, RGB, grayscale,
HSV, or CIE L*a*b* . Depending on the color and hue of the image analyzed, the total
value or just one channel of the color space can be used for quantification. Usually the
quantitative response is given as the arithmetic mean of the pixel intensity within the
detection zone after subtraction of the mean intensity for the control region.
Another interesting group of detectors incorporates simple photoelectric meters
(Şaki, N., 2004) and homemade devices. Several research groups have already
presented small devices that can be fabricated with off-the-shelf components, e.g., the
portable color identifier described by Li et al. consisting of light-emitting diodes (LED),
a photodiode, an integrated chip, a power source, an amplifier, and capacitors or the
transmittance colorimeter described by the Whitesides group, both assembled from
components readily available in any electronics supply catalog. Not so common, but
still useful options are spectrophotometers and fluorometers that could be applied to
optimize the fabrication process in the laboratory or in their portable versions for on-site
 7

detection. Finally more sophisticated equipment can be used, such as microplate

readers, photomultiplier tubes, and gel documentation systems, but these kinds of
devices are rather expensive, not so popular, and therefore not fully compatible with
simple paper-based sensors (Tunç, S., 2017).

2.2. Colorimetric methods

Colorimetric paper-based sensors are surely the most prevalent as this substrate
offers a bright, high-contrast, colorless background for color change readings. Sensors
of this type are so numerous that their review will be divided into groups (Zhou et al.,
2013). For example, The first work on paperbased devices from the Whitesides group—
so often cited as the breakthrough in paper-based diagnostics—describes an assay for
glucose and proteins. In this two-dimensional device channels are delimitated with
photoresist and detection zones are modified with tetrabromophenol blue for protein
detection and a potassium iodide, horseradish peroxidase, and glucose oxidase mixture
for glucose detection. The detection range is 0.38–7.5 mM for bovine serum albumin
and 2.5– 50 mM for glucose (Georgakilas et al., 2012).

2.3. Fluorescence methods

Two flaws of paper-based fluorescence are the need for additional

instrumentation and the fact that many commercial papers fluoresce (because of
additives to increase the impression of whiteness), resulting in high background signal.
Nevertheless, many paper-based fluorescence sensors have been developed and have
achieved low limits of detection (Ferrari et al., 2015). An interesting example is the
sensor described by Thom et al. They presented a sensor for β-Dgalactosidase and
albumin integrated with a galvanic cell battery, all made from paper. The battery serves
as a power source for the UV LED used to illuminate the assay region. This galvanic
cell consists of several components incorporated in interchangeable layers of paper and
tape. Square or disk metal electrodes are incorporated into air gaps in the patterned
layers of tape, salt bridges and electrolytes are predeposited in hydrophilic regions, and
finally conductive connections are made from copper tape. Batteries can be fabricated
from a range of materials and they can be connected in series or in parallel, resulting in
the desired current or potential value.
 8

Figure 2.1 β-D-galactosidase assay with integrated light-emitting diode (LED) and fluidic
battery

2.4. Chemiluminescence methods

Chemiluminescence methods are based on measurement of the intensity of light

emitted by a chemical reaction. They are generally praised for their simplicity, high
sensitivity, low cost, and compatibility with micromachining technologies as well as the
fact that the measurement may be performed in the dark (Zor et al., 2015).
The next device is an example of electrochemiluminescence detection where
light is produced as a result of an electrochemical reaction on the electrode. Delaney et
al. presented a platform for quantification of NADH and 2-(dibutylamino)ethanol.
Graphite working and auxiliary electrodes and a silver reference electrode were
fabricated on a plastic substrate. A hydrophilic channel on paper was delimitated with
alkenyl ketone dimer by means of an ink-jet printer. Tris(2,20-bipyridyl)ruthenium(II)
was deposited on the patterned paper. The paper layer was stacked on a plastic sheet
with screen-printed electrodes and laminated. A small incision was made to allow
introduction of the sample. The reaction was initiated when the potential of the working
electrode was scanned by the oxidation potential of the ruthenium complex, resulting in
orange luminescence.
 9

Figure 2.2. Assay for simultaneous quantification of glucose and uric acid

2.5. Detectors

Apart from electrochemical workstations available in the laboratory, several

simple detector systems have been proposed to permit field measurements. The first
group includes homemade devices that can be fabricated from off-the-shelf
components (an amplifier, a voltage regulator, a voltage inverter, batteries). They
are characterized by their simplicity and low cost, and could be adapted to work on
batteries or to draw power from the network (Hyun et al., 2013). They can be
assembled at the place of need, which is important for applications in developing
countries. Multimeters form the next class of potential electrochemical detectors.
They are readily available and affordable (approximately $30) and have already
been integrated in iPhones and iPods as socalled iMultimeters(Huang et al., 2011).
Commercial glucose meters are another group of detectors compatible with
paper-based platforms. They are designed to be operated by unskilled personnel.
The cost of the development of this technology has already been absorbed by the
market; thus, we can take advantage of this sophisticated engineering for a
reasonable price. Assays can be adapted to a variety of analytes and types of
measurement, such as amperometry, chronoamperometry, anodic stripping
voltammetry, and cyclic voltammetry.
 10

3. FABRICATION AND PATERNING

In fabricating paper devices, the choice of techniques and materials that meet the
criteria of low cost, simplicity and efficient production process need to be considered.
There are several techniques and processes involving chemical modification and/or
physicaldeposition that could be used to tune the properties of the paper such thatit
becomes available for further modification or direct usage in a range of applications
(Ramsden, J., 2009).
Techniques reported in the literature include photolithography, analogue
plotting, inkjet printing and etching, plasma treatment, paper cutting, wax printing,
flexography printing, screen printing, and laser treatment. Techniques were chosen
depending on the typeof material used and the typeof modification required. Much
research is focused on confining the liquid to a specific region on the paper, in whatis
known as paper-based microfluidics, so first we discuss some of these approaches
followed by some other methods to build up the active sensing element (Huang et al.,
2011).
In 2007, Martinez et al. introduced a lithographic technique to create a
microfluidic channel by using a hydrophobic photoresist, SU-8 polymer (Figure 3.1).
The hydrophilic channel defined the liquid penetration pathway as it was confined
within the hydrophobic walls. As the liquid was introduced to the hydrophilic channel,
it moved through the paper matrix by capillary flow action. A three-branch tree pattern
was lithographically patterned on the paper for the reaction site where different reagents
were spotted for glucose and protein assays. This work was a major breakthrough that
led to significant research growth in this field. It is attractive as it offers a simple and
relatively inexpensive alternative to existing technologies and is suitable for portable
applications. Since the pioneering work by Martinez et al. on paper-based microfluidics,
alternative approaches have been introduced by other researchers to create a hydrophilic
channel confined within a hydrophobic barrier. Physical deposition of patterning agents
such as wax, polydimethylsiloxane and polystyrene have been used to create paper
devices by many research groups. One potential problem with the use of
photolithography to create a microfluidic channel is that damage of the photoresist may
occur during bending or folding. To overcome the problem introduced by
photolithography, Bruzewicz et al. demonstrated printing of elastomeric
polydimethylsiloxane onto paper using a plotter. This allowed folding of the paper
 11

device without destruction of the channel. Moreover, the technique is highly

reproducible and usesinexpensive materials, and is thus suited for even the most basic
of research laboratories where microfluidic devices withfeature sizes of ~1 mm are
adequate.

Figure 3.1. (a) Steps involved in fabricating paper with millimetre-sized channels using
photolithography and (b) spotting of the paper for glucose and protein assays

3.1. Inkjet printing

Rapid, precise, and reproducible deposition of a broad variety of functional

materials, including analytical assay reagents and biomolecules, has made inkjet
printing an effective tool for the fabrication of microanalytical devices. A ubiquitous
office device as simple as a standard desktop printer with its multiple ink cartridges can
be used for this purpose.
In terms of application, inkjet technology is divided into two major categories:
drop-on-demand (DOD) and continuous inkjet (CIJ). All of the common desktop
printers and all laboratory-relevant inkjet printing systems rely on DOD technology. In
contrast to the CIJ mechanism with a continuous ejection of fluid, the jetting of ink onto
the substrate only occurs when it is needed, as implied by the term “drop-ondemand”.
DOD inkjet printing is divided into four modes of actuation: piezoelectric, thermal,
electrostatic, and acoustic. At present, only the piezoelectric and thermal methods are
routinely utilized. Figure 3.2 demonstrates the process of fluid jetting for these two
 12

DOD printers. Initially, the ink inside the cavity experiences an increase in pressure
arising from a piezoceramic deformation (piezoelectric) or a vapor bubble expansion
(thermal). Rapid expansion of a vapor bubble is achieved by superheating the ink by a
heat pulse applied for about 2 µs (Morales-Narváez et al., 2013).

Figure 3.2. . Basic working principle of drop-on-demand (DOD) inkjet

printing: a) piezoelectric inkjet and b) thermal inkjet.

Major methods for patterning channels in mPADs are briefly discussed, with the
exception of inkjet printing, to which the following Section 4 is devoted. Figure 3.3
gives an overview of various channel fabrication principles. In the simplest case,
patterning can be achieved by cutting the paper which is not further discussed here. A
more widely used approach is treatment with a hydrophobic material (Figure 3.3). The
hydrophobic modification approaches can be subdivided into indirect and direct
patterning methods. Indirect patterning methods rely on the use of a mask to project the
shape of the microfluidic channel structure on the paper. On the other hand, direct
methods involve the deposition of hydrophobic compounds onto selected positions of
the paper substrate in such a way as to define the structure of the flow channel. The
main objective in the fabrication of microfluidic channels on paper substrates by
hydrophobic treatment is to obtain a contrast between the hydrophobic barrier and the
hydrophilic flow channel. The hydrophobically impregnated areas define the
 13

dimensions of the channel (Figure 3.3). The porosity and surface composition of
cellulose fibers are two important characteristics for the covalent or noncovalent
hydrophobic treatment of the substrate. Porosity allows the penetration and filling with
liquid through the void spaces (Lee et al., 2010).

Figure 3.3. Methods to create microfluidic channels on paper substrates: a) mechanical

cutting of paper and b) treatment with hydrophobic materials.

3.2. Photolithography

Photolithography is the process of transferring geometric shapes on a mask to

the surface of a silicon wafer. The steps involved in the photolithographic process are
wafer cleaning; barrier layer formation; photoresist application; soft baking; mask
alignment; exposure and development; and hard-baking.
In the first step, the wafers are chemically cleaned to remove particulate matter
on the surface as well as any traces of organic, ionic, and metallic impurities. After
cleaning, silicon dioxide, which serves as a barrier layer, is deposited on the surface of
the wafer. After the formation of the SiO2 layer, photoresist is applied to the surface of
the wafer. High-speed centrifugal whirling of silicon wafers is the standard method for
applying photoresist coatings in IC manufacturing. This technique, known as "Spin
Coating," produces a thin uniform layer of photoresist on the wafer surface (Kratzer et
al., 2015).
There are two types of photoresist: positive and negative. For positive resists, the
resist is exposed with UV light wherever the underlying material is to be removed. In
these resists, exposure to the UV light changes the chemical structure of the resist so
 14

that it becomes more soluble in the developer. The exposed resist is then washed away
by the developer solution, leaving windows of the bare underlying material. In other
words, "whatever shows, goes." The mask, therefore, contains an exact copy of the
pattern which is to remain on the wafer. Negative resists behave in just the opposite
manner. Exposure to the UV light causes the negative resist to become polymerized,
and more difficult to dissolve. Therefore, the negative resist remains on the surface
wherever it is exposed, and the developer solution removes only the unexposed
portions. Masks used for negative photoresists, therefore, contain the inverse (or
photographic "negative") of the pattern to be transferred. The figure below shows the
pattern differences generated from the use of positive and negative resist (Liu et al.,
2015).
Negative resists were popular in the early history of integrated circuit
processing, but positive resist gradually became more widely used since they offer
better process controllability for small geometry features. Positive resists are now the
dominant type of resist used in VLSI fabrication processes (Chan et al., 2016).

Figure 3.4. Schematic of the photolithography technique for fabricating paper-based

microfluidic platforms: (a) procedure for patterning paper with hydrophobic
photoresist, (b) derivatization of the device for bioassays.
 15

Soft-baking is the step during which almost all of the solvents are removed from
the photoresist coating. Soft-baking plays a very critical role in photo-imaging. The
photoresist coatings become photosensitive, or imageable, only after softbaking.
Oversoft-baking will degrade the photosensitivity of resists by either reducing the
developer solubility or actually destroying a portion of the sensitizer. Undersoftbaking
will prevent light from reaching the sensitizer. Positive resists are incompletely exposed
if considerable solvent remains in the coating. This undersoft-baked positive resists is
then readily attacked by the developer in both exposed and unexposed areas, causing
less etching resistance (Huang et al., 2011).
One of the most important steps in the photolithography process is mask
alignment. A mask or "photomask" is a square glass plate with a patterned emulsion of
metal film on one side. The mask is aligned with the wafer, so that the pattern can be
transferred onto the wafer surface. Each mask after the first one must be aligned to the
previous pattern. Once the mask has been accurately aligned with the pattern on the
wafer's surface, the photoresist is exposed through the pattern on the mask with a high
intensity ultraviolet light. There are three primary exposure methods: contact,
proximity, and projection. They are shown in the figure below. Hard-baking is the final
step in the photolithographic process. This step is necessary in order to harden the
photoresist and improve adhesion of the photoresist to the wafer surface(Liu et al.,
2015).
 16

4. TRENDS OF PAPER BASED OPTICAL SENSORS

Since the beginning of the μPAD development the right selection of the shape
and the geometry on paper has been critical to achieve devices with high performance.
The important role of geometry in the paper microfluidic devices is inherent since the
Washburn and Darcy models describe the hydrodynamic processes in porous paper
considering the channel length and its transverse area. A good selection of the paper
shape can enhance the sensitivity, the multianalysis capacity and the multifunctionality
of the devices. A lot of works are focused on the change in the conventional geometry
of the paper strips to introduce larger volumes of sample, to concentrate the analyte
sample, to provide the multiple analytes analysis and to reduce the steps in the reagent
incorporation and consequently the time of the analysis.

4.1. 2D and 3D μPADs

The use of several paper layers or paper folds enhanced the ability of the devices
to generate interesting uses. These devices are called 3D μPADs. They distribute fluids
both vertically and laterally, and they enable streams of fluid to cross one another
without mixing. These devices use capillary wicking to distribute fluids into complex
arrays of multiplex detection zones in times of seconds to minutes (depending on the
architecture of the device and the choice of materials), permitting multianalysis in one
sample or sample multianalysis. Their architecture made by several stackable layers
may be moved, added, or removed to enable a desired function. The capability of the
3DμPADs to incorporate complex networks of channels provides them multiple
functionalities (serving as a substrate for filtering samples, performing chromatographic
separations, and taking biochemical reactions) (Hyun et al., 2013).
3D µPADs are fabricated by stacking alternating layers of paper and water-
impermeable double-sided adhesive tape (both patterned in ways that channel the f low
of f luid within and between layers of paper). The hydrophobic polymer patterned into
the paper demarcated the channels through which the f luids moved laterally (8, 9), the
layers of water-impermeable doublesided tape separated channels in neighboring layers
of paper, and holes cut into the tape allowed f luids to f low vertically.
The layers of paper were patterned with SU-8 2010 photoresist. Paper was
impregnated with photoresist, dried, exposed to UV light through a transparency mask,
and developed with acetone and isopropyl alcohol (8, 9). The layers of tape were
 17

patterned using a laser cutter. The tape could also be patterned manually, using a hole
punch, albeit much more slowly (Turner et al ., 1987).. The protective film was
removed from the tape, and a second layer of paper was aligned and attached to its
second surface. This process of stacking—paper, tape, paper, tape—was repeated as
needed to complete a device, and provided a reproducible method for fabricating
devices. The cellulose powder was used to fill the gaps between adjacent layers of paper
that were created by the thickness of the tape. Without the cellulose powder, fluids did
not wick reproducibly between two adjacent layers of paper (Guo et al., 2010).

Figure 4.1. Preparation and demonstration of a 3D µPAD.

 18

4.2. Modifications to the classical Lateral Flow Assays format

The classical LFA format has also been modified in order to provide
advantageous devices retaining the positive aspects of conventional LFAs. One of the
first reports of 2D geometry was made in 2006 by J.-H. Cho modifying the classical
LFIA format. In this work a 2D LFA that used a HRP enzyme as a signal generator
colour was placed into channels etched on the plastic chip surface for cTnI analysis. The
chip consisted of two cross-flow channels in the lateral and transverse directions. In the
lateral channel, they placed a 2 mm wide immunostrip for analyte analysis, which was
identical to a conventional LFA. In the transverse direction, an enzyme substrate supply
channel and a flow absorption pad compartment were arranged on each lateral side of
the signal generation pad of the lateral strip (Baptista-Pires et al., 2016).

Figure 4.2. Construction of an analytical chip for ELISA adapting the concept of cross-flow
chromatography.
 19

4. MARKET OF PAPER BASED OPTICAL SENSORS

4.1. Market of optical sensors

The global automation industry, particularly the process automation and industry
is growing at around 6.3% and they are the primary growth drivers for the global optical
sensors market.
According to the latest market research report “Optical Sensors Market
Analysis: By Types (Position, Light, Image, Photoelectric, Photodetectors, Motion);
Verticals (Material Handling, Building Automation, Manufacturing, Packaging,
Security & Surveillance) – With Forecast (2015-2020)”, published by IndustryARC,
the global optical sensor market is slated to grow at 9.81% CAGR year on year for next
5 years to reach $34billion by 2020.
Optical sensors are electronic detectors which work on the presence or absence
of light or infrared rays and convert the same into an electronic signal. Usually they are
part of a much larger system where this electronic signal is interpreted or analyzed in
different ways to produce a result like presence of a person, capturing an image,
movement of an object or position of an object and so on (Singh ve ark., 2017).
The Global Optical Sensor Market is rapidly growing across the globe due to the
increase in demand for the handheld devices like tablets and smart phones. Smart
phones and Tablets are the key growth contributors and as long as the market persists
there will be increase in demand for optical sensors like light and proximity sensors. On
the other hand image sensor market is skyrocketing compared to other sensors as it’s
very widely used. The Market for image sensors goes from cameras to medical
applications. The wide market for image sensors is making it the largest segment in the
optical sensor industry. The Global Optical Sensor Market is slated to grow at 9.81%
CAGR year on year to reach $34 Billion by 2020 (Malzahn et al., 2011).
The global automation industry, particularly the process automation and industry
is growing at around 6.3% and they are the primary growth drivers for the global optical
sensors market. Increasing use of automation in different industry verticals like food
and beverage, chemicals, petrochemicals, paper and pulp, packaging drives the need for
installation of products or hardware which uses optical sensors on a larger scale. Image
sensors are used in machine vision based products for assembly line monitoring and
other automation purposes. Color sensors for example are used in the packaging, food
 20

and beverage industry for accuracy purposes. Compared to image sensors they are
cheaper, faster and can be replaced easily.
Optical Sensors have created a growing attention in health monitoring through
various applications that can be integrated with wearable gadgets which include
watches, smart phones and fitness equipment. Optical sensors enlarge the opportunities
through self-observation and also provide features like pulse rate, oxygen levels and
saturation levels through small gadgets for wrist and fingers which is more convenient.
Wearable optical sensors manufactured with plastic semiconductors are now
currently emerging into the market as low cost and disposable products. Wearables are
significant for regular monitoring and the main advantage is the product is of light
weight, flexible and easy to manufacture. These are light-based sensors which can be
used to monitor the changes in the body by detecting the muscle contraction and
expansion accordingly. A very stimulating conceivable use of the optical sensor is the
process of monitoring the prosthetic limbs and possibly artificial heart without any
insertion of needles into the body. The optical sensors technology in health care with
various portable devices is driving the medical tests from hospital to home. The various
health monitoring wearable devices that are available in the market are limited to being
fitness devices which would definitely need sprucing up for use in Healthcare and
Medicine.

Figure 5.1. Expectation for sensor market by 2022.

 21

4.2. Market of paper based optical sensors

Paper-based sensors have also found applications in industry from the early
stages of their development. This simultaneous development in academic/research labs
and the industrial commercialization of PADs has been justified by numerous
advantages, with the simplicity and low cost of the mass production implementation, the
increasing necessity to have an affordable POC sensor that is easy to use by any person
through telemedicine and in field issues, and the robustness and accuracy of the devices
being some of the principal factors responsible for this rapid technological transfer.
The first paper-based bioassay was introduced in 1957 for the identification of
Glc in urine. The development of a blue colour on the strip indicated abnormal Glc
levels. In the middle of the 1960s, this assay was developed into a commercial product
to diagnose and manage diabetes. In the same year a triple-test dipstick to detect urinary
Glc, albumin, and pH involving three different reagent-deposited zones (“patches”) and
developing distinct colours as analyte response was introduced by H. M. Free et al.
After that, 10-test urinary dipsticks including other analytes such as leukocytes, nitrites,
ketones, urobilinogen, bilirubin, specific gravity and blood were developed and
commercialized. These dipsticks were packaged with a colour-coded chart that allowed
the user to read out the detected quantity of a given analyte at a defined location along
the length of the strip similar to a paper pH reader, which was widely accepted by the
medical community. Today, commercial urinalysis dipsticks have been widely adapted
for several analytes. Such is the case of URiSCAN Strips (for 12 urine analytes),
Chemstrip®(for Glc), Multistix®10SG Reagent Strips (10 tests for important markers
for diabetes and kidney disorder) and CLINITEK Microalbumin 9 Reagent Strips (9
tests for managing patients with diabetes and kidney disease), which present a
colorimetric chart for the test interpretation. Additionally, reader strip equipment has
been developed too.
In 1964 the Ames group greatly improved the dry reagent dipstick for testing
blood Glc, developing an enzymatic blood Glc assay that was sold as Dextrostix. This
filter paper strip in addition to containing the dried reagents (GOx–HRP– chromogen
reaction mixture) has a clear semipermeable barrier easing diffusion of the plasma in the
drop of blood into the reaction mixture of the filter paper and producing the colour
development. This prevented blood interferences and opened up the possibility for an
instrument reader. In 1970 Ames introduced the first hand-held battery-operated
 22

reflectance meter called ARM™. In the scientific literature the first immunotest strip
was presented in 1982 for the assay of Abs in seravia the application of the Ag as a spot
to nitrocellulose filters. In 1983 urine-based home-use immunoassay test kits for various
bioanalytes, including hCG and 9-tetrahydrocannabinol (pharmacological agent of
marijuana) were commercially available. Blood-based home-use immunodipsticks for
Chol, diabetes, hepatitis C and HIV-1 have been in common use in the clinical
laboratory for screening of diseases since the 1980s.
Although dipstick and LF formats have dominated the rapid diagnostics market
over the past three decades, recently, some examples of commercial μPAD applications
have emerged as versatile POC platforms transcending the capabilities of existing
assays. The industrial transfer of some of these devices is still a challenge, in spite of
the numerous μPAD advantages, because there are multiple factors defining the
commercialization strategies. Nevertheless, some companies have taken advantage of
the variety of laboratory works and academic proof-of-concept studies based on LOCs
and μPADs with attractive advances over the classical formats.
Many of these interesting applications have been fortunately introduced into the
market by companies currently developing the next generation of technologies. For
example Alere, Abaxis, Abbott, Achira labs, Agilent, Biosite, Chembio and Daktari
Diagnostics have produced devicesviathe integration of microfluidic and LOC
technologies with paper-based assays.
 23

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