DOI: 10.1002/asia.

201700814 Focus Review

Sensor Design

Fluorescence and Sensing Applications of Graphene Oxide and

Graphene Quantum Dots: A Review
Peng Zheng and Nianqiang Wu*[a]

Chem. Asian J. 2017, 12, 2343 – 2353 2343 T 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Abstract: Graphene oxide and graphene quantum dots are
attractive fluorophores that are inexpensive, nontoxic, pho-
tostable, water-soluble, biocompatible, and environmentally
friendly. They find extensive applications in fluorescent bio-
sensors and chemosensors, in which they serve as either flu-
orophores or quenchers. As fluorophores, they display tuna-
ble photoluminescence emission and the “giant red-edge
effect”. As quenchers, they exhibit a remarkable quenching
efficiency through either electron transfer or Fçrster reso-
Introduction
Graphene-based nanomaterials are two-dimensional (2D)
atomic crystals composed of sp2-hybridized carbon atoms.[1]
The family includes graphene, graphene oxide (GO), reduced
graphene oxide (rGO), and graphene quantum dots (GQDs).
Ideally, graphene is completely composed of sp2-hybridized
carbon atoms, which exhibit no fluorescence due to a zero op-
tical bandgap. Due to the introduction of functional groups,
GO, rGO, and GQDs consist of mixtures of sp2 and sp3 carbons
that open the optical bandgap and result in fluorescence.[2] GO
is rich in oxygen-containing functional groups. The chemical
reduction of partial oxygen-containing functional groups in GO
leads to the formation of rGO.[3] GQDs are cut from a single to
several atomic layers of graphene sheets several nanometers in
lateral size.[4] Typically, GQDs contain both sp2 and sp3 carbon
atoms, which are inherent from synthesis processes. GQDs
have a size-dependent optical bandgap due to the quantum
confinement.[5] GO and GQDs are inexpensive, nontoxic, photo-
stable, water-soluble, biocompatible, and environmentally
friendly, and they exhibit excitation-wavelength-dependent
fluorescence.[6] In addition, their 2D surface shows strong non-
covalent interactions with adsorbed biomolecules through p–p
interactions, electrostatic forces, or hydrogen bonding, which
provide a chemically tunable platform for bioconjugation.[6–7]
Therefore, both GO and GQDs have found extensive applica-
tions in fluorescent sensors in recent years. They can be em-
ployed not only as a tunable fluorophore but also as an effec-
tive fluorescence quencher.
We begin with a discussion of the physical principles of fluo-
rescence of GO and GQDs. We then show how to design fluo-
rescent sensors by utilizing the unique physicochemical prop-
erties of GO and GQDs. Also, we highlight the applications of
GO- and GQD-based fluorescent sensors in a variety of fields.
[a] P. Zheng, Prof. N. Wu
Department of Mechanical and Aerospace Engineering
West Virginia University
PO Box 6106, Morgantown, WV 26506 (USA)
E-mail: nick.wu@mail.wvu.edu
The ORCID identification number(s) for the author(s) of this article can be
found under https://doi.org/10.1002/asia.201700814.
Chem. Asian J. 2017, 12, 2343 – 2353 www.chemasianj.org
Focus Review
nance energy transfer (FRET) process. In this review, the
origin of fluorescence and the mechanism of excitation
wavelength-dependent fluorescence of graphene oxide and
graphene quantum dots are discussed. Sensor design strat-
egies based on graphene oxide and graphene quantum
dots are presented. The applications of these sensors in
health care, the environment, agriculture, and food safety
are highlighted.
Fluorescence Principles of Graphene Oxide
Fluorescence Origin
Electronic energy transitions are responsible for the fluores-
cence of GO.[5] As shown in Figure 1, each ’’fingerprint’’ fluores-
cence band of functionalized GO comes from specific electron-
ic transitions between the antibonding and the bonding mo-
lecular orbitals such as s*!n, p*!n, and p*!p.[5] GO general-
ly contains various oxygen-containing functional groups, such
as hydroxyl groups (COH), carboxyl groups (COOH), carbonyl
(C=O), epoxy (C-O-C), and aromatic rings (C=C). As a result,
multiple fluorescence peaks, which correspond to different
electronic transitions, are usually excited simultaneously. When
these fluorescence peaks overlap with each other, a single
broad peak is observed, as shown in Figure 1.[8]
Functional groups, dopants, lateral size, localized domains,
and strain in GO can alter the electronic energy transitions and
lead to changes in the fluorescence peak position, width, and
shape. For example, after GO is incubated in aqueous KOH or
HNO3, it becomes enriched with OH or COOH groups, respec-
tively. Consequently, the fluorescence peak position and inten-
sity are tuned as shown in Figure 1.[8] Bear in mind that the
bandgap of GO is created as a result of the mixture of sp2 and
sp3 carbon atoms after the introduction of functional groups.
The electronic band structure can be tuned by varying the
ratio of sp2/sp3 carbon atoms, which allows the fluorescence to
be tuned from the visible light to the near-infrared (NIR) wave-
length range. The fluorescence of GQDs has the same origin as
GO. Doping has been found to modulate the electronic struc-
ture of GQDs and thus affect the fluorescence emission. For ex-
ample, in nitrogen-doped GQDs, the fluorescence is dominated
by the n!p* transition between the aromatic rings that con-
tain N and the conjugate structure of graphene.[9] An increase
in the doping percentage of nitrogen in GQDs could redshift
the emission and improve photostability.[10] Due to quantum
confinement, changes in the size of GQDs can modulate the
electronic structure and thus shift the fluorescence from green,
to red, to NIR. In addition, changes in the localized domains, in
which p electrons are confined in localized sp2 regions, can
also alter the fluorescence of GO.[10–11]
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Figure 1. a) UV/Vis absorption of as-synthesized GO and GO treated with KOH and HNO3. b) Fluorescence spectra corresponding to a); inset: the different
electronic transitions. Reproduced from Ref. [8] with permission from The Royal Society of Chemistry.
Excitation-Wavelength-Dependent Fluorescence
Breakdown of Kasha’s rule
According to Kasha’s rule, the peak fluorescence wavelength
for organic dyes and inorganic quantum dots is independent
of the excitation wavelength because fluorescence always
starts at the band edge after rapid internal relaxation. Howev-
er, Kasha’s rule does not hold for GO. Excitation-wavelength-
dependent fluorescence occurs in GO suspended in polar sol-
vents, as shown in Figure 2.[8] When the excitation wavelength
was increased from l = 325 to 650 nm, the fluorescence band
was redshifted to longer wavelengths.
Figure 2. Excitation-wavelength-dependent fluorescence of GO. Reproduced
from Ref. [8] with permission from The Royal Society of Chemistry.
Giant red-edge effect
Excitation-wavelength-dependent fluorescence of GO is due to
the “giant red-edge effect”.[12] As shown in Figure 3, the giant
red-edge effect occurs in a polar solvent, such as water, in
which the solvation introduces an additional relaxation step.
For an excited fluorophore in a polar solvent, the dipole is not
in equilibrium with the solvent dipole. During solvation, these
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Focus Review
dipoles are realigned with one another and reduce the interac-
tion energy to achieve equilibrium. For a common fluorophore
in a polar solvent, the solvation dynamics are orders of magni-
tude faster than fluorescence. The solvation is usually complet-
ed prior to fluorescence. Therefore, the final fluorescence only
undergoes a small redshift. However, if the solvation dynamics
are slowed down to the fluorescence timescale, fluorescence
occurs simultaneously as the energy of the excited fluorophore
is reduced, which leads to a strong dependence of fluores-
cence on the excitation. The giant red-edge effect takes place
not only in GO but also in GQDs and carbon dots.
Mr. Peng Zheng received his B.S. degree in
2011 from Central South University, China,
and his M.S. degree in 2014 from West Virgin-
ia University (WVU), USA. He is working
toward his Ph.D. degree at WVU in the group
of Prof. Nianqiang Wu. He strives to under-
stand the optical properties of nanostructures
and to develop biosensors.
Dr. Nianqiang (Nick) Wu is currently Professor
of Materials Science at West Virginia Universi-
ty, USA. He is a Fellow of Royal Society of
Chemistry (FRSC) and a Fellow of the Electro-
chemical Society (FECS). He serves on the
Board of Directors in the Electrochemical Soci-
ety (ECS) and is Chair of Sensor Division. His
research interest lies in photocatalysts and
photoelectrochemical cells, batteries and su-
percapacitors, and biosensors and labs-on-
chips.
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Figure 3. Fluorescence properties of GO. GO exhibits excitation-wavelength-
dependent fluorescence (or giant red-edge effect) due to solvation. It also
displays an excited-state protonation from COOH groups. The measured
fluorescence of GO is a superposition of contributions from COH and COOH
groups. Reprinted with permission from Ref. [12]. Copyright (2014) American
Chemical Society.
Excitation-wavelength-dependent fluorescence due to strain
Given the fast solvation dynamics ( & 10@12 s@1) of the polar sol-
vents used, there must be some important factors relevant to
material itself that drastically slow down the dipole realign-
ment process to the timescale of fluorescence. In this regard,
efforts have been made to study the effects of oxygen per-
centage, doping percentage, disorder, and strain in GO, which
have been commonly hypothesized to be the underlying rea-
sons for the giant red-edge effect.[11a, 13] Of the aforementioned
factors, strain has been found to correlate with the giant red-
edge effect.[14] When there is an out-of-plane strain, the plane
of the GO sheet becomes curved, which generates a strong
local reorganization potential. The local potential prevents a
fast reorientation of the excited dipoles with the solvated di-
poles in the oxygen-containing functional groups nearby and
drastically slows down the dipole realignment process to the
fluorescence timescale, which generates the giant red-edge
effect.[14] The redshift in the peak fluorescence wavelength in
GO is also found to scale linearly with the increase in the exci-
tation wavelength.[8, 12] The peak fluorescence can be tuned
from visible to the NIR region by simply changing the excita-
Figure 4. pH-dependent photoluminescence of GO excited at l = 440 nm. a) The pH effect on photoluminescence. b) Plot of the photoluminescence intensi-
ties at l = 683, 506, and 479 nm with respect to the pH of the GO solutions. Reprinted with permission from Macmillan Publishers Ltd: [Scientific Reports]
(Ref. [11a]), copyright (2011).
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Focus Review
tion wavelength without changing any other factor, such as
chemical composition and size.
Influence of pH on GO fluorescence
In addition, the fluorescence of GO is dependent on the pH
value of the solvents, as shown in Figure 4.[11a] Under acidic
conditions, GO exhibits a broad fluorescence peak centered at
around l = 680 nm. As the pH gradually increased, the l =
680 nm peak reduced in intensity, and finally disappeared with
the emergence of two new peaks at around l = 500 nm under
basic conditions. This pH-dependent fluorescence is believed
to be caused by an excited-state proton transfer, as shown in
Figure 3. Under acidic conditions, the COOH group is largely
deprotonated and exists in the form of ionic COO@ in the
ground state. Upon excitation, the excited-state protonation
from ionic COO@ to COOH contributes to the broad fluores-
cence peak at l = 668 nm. In comparison, the fluorescence
emission under basic conditions is associated with the excited
COO@ moieties.
Sensing Applications of GO and GQDs
GO can be used as a fluorophore with tunable fluorescence
emission by varying the chemical composition, size, and other
factors. GO can also be used as a highly efficient quencher
through either charge-transfer or resonance-energy-transfer
processes owing to its super electrical conductivity and 2D
planar structure. The dual roles of GO as both a fluorophore
and a quencher open up new opportunities for sensor design,
as shown in Figure 5. Therefore, graphene-based nanomaterials
are finding increasing applications in environment, agriculture,
biomedicine, food safety, and homeland security. This section
outlines different design strategies for fluorescent biosensors
and also highlights some important applications.
GO as a fluorescent label
GO has been widely used as a fluorescent label, just like organ-
ic dyes and inorganic quantum dots.[15] However, the broad
emission peak limits the sensing performance when the fluo-
rescence spectral intensity is transduced as the signal. Instead,
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Figure 5. Schematic illustration of sensor design based on GO. a) GO as a fluorescent label. b) GO as a FRET donor and its fluorescence can be quenched by a
quencher, such as a gold nanoparticle, through FRET. c) Charge transfer occurs between GO and a fluorophore in close proximity. d) GO acts as a quencher
and can quench the fluorescence of a nearby fluorophore through FRET.

GO is remarkably suitable for NIR biological imaging by using
two-photon excitation spectroscopy thanks to the giant red-
edge effect.[12] Two-photon excitation is based on the simulta-
neous absorption of two photons that have only half the
energy of a single photon traditionally used to excite the same
event.[16] The selection of a light source with a longer wave-
length, particularly NIR excitation, greatly helps to reduce pho-
totoxicity, lower the background signal, and increase the light
penetration depth in a biological sample. GO with excitation-
wavelength-dependent fluorescence is well coupled with two-
photon excitation spectroscopy because its excitation wave-
length can be deliberately chosen to be fallen into one of the
biological windows to allow selective biological imaging.[17]
The first and second NIR biological windows are located at l =
650 to 950 and 1000 to 1350 nm, at which biological samples
are almost transparent to incident light.[18] As an example, GO
exhibits different fluorescence when excited in the first and
second biological windows by using two-photon excitation
fluorescence spectroscopy, as shown in Figure 6a and b.[17] The
advantages of the use of GO in biological imaging under two-
photon excitation are further manifested by applications in
cancer treatment, as shown in Figure 6c–f.[19] GO nanoparticles
(GONs) were functionalized with transferrin (Trf) molecules to
target the cancer cells. The Trf-GO needs a much lower laser
power to generate sharp and well-resolved GON images. Be-
cause Trf can specifically target cancer cells, so does the Trf-
GON complex. The process of killing cancer cells by using the
Trf-GON complex is thus directly reflected in the GON fluores-
cence images. Liu et al. have also reported cellular and deep-
tissue imaging by using two-photon fluorescence spectroscopy
based on GQDs.[20] Wang et al. took advantage of two-photon
spectroscopy to image endogenous biological cyanide in plant
tissues based on a GQD–gold nanoparticles complex.[21] Two-
photon imaging with GO as a fluorescent label has been em-
ployed in biomedical fields to target liver tumor cells,[22] breast
tumor cells,[23] melanoma cells,[24] and also for food safety.[21]
GO as a donor in charge transfer
The fluorescence of GO can be quenched by a charge-transfer
process, in which GO acts as a charge donor.[25] Compared to
FRET, which is a long-range energy transfer mechanism, charge
transfer is a short-range energy transfer process. It requires the
charge donor and the acceptor to be separated by within

Figure 6. Two-photon fluorescence imaging a) tunable excitation-wavelength-dependent two-photon imaging in the first and second biological transparency
windows. b) Two-photon fluorescence imaging of methicillin-resistant Staphylococcus aureus (MRSA) by using aptamer-modified GO, in which the excitation
wavelength for b1)–b4) were l = 1160, 880, 980, and 760 nm, respectively. Reprinted by permission from Macmillan Publishers Ltd: [Scientific Reports]
(Ref. [17]), copyright (2014). c)–f) A comparison of in vitro two-photon luminescence imaging of GO nanoparticles and molecular dye FITC. Reprinted from
Ref. [19] with permissions from Wiley.

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 Focus Review

Figure 7. Fluorescent sensors based on charge transfer. a) Dopamine detection based on charge transfer between GO and dopamine through p–p stacking.
Reprinted with permission from Ref. [15c]. Copyright (2011) American Chemical Society. b) Hg2 + detection, in which Hg2 + is captured through a thymine-
Hg2 + -thymine (T-Hg-T) interaction and quenches the fluorescence of GO. Reprinted from Ref. [25], Copyright (2013), with permission from Elsevier. c) Hg2 +
and I@ can both be detected on the same GO fluorescent biosensor platform. Reprinted with permission from Ref. [27]. Copyright (2015) American Chemical
Society.

10 a.[26] Such a short distance is often realized through chemi-
cal bonding or physical adsorption. As an example, dopamine
molecules have been detected in a GO fluorescent sensor
based on charge transfer (Figure 7a.[15c] The dopamine mole-
cule is chemically bonded to GO through the p–p interaction
between GO and the aromatic rings in the molecules. In the
excited GO–dopamine complex, the excited electrons in GO
are transferred to the dopamine molecule, which results in
quenching of the GO fluorescence. In addition, Wu’s research
group used the charge-transfer mechanism to detect metal
cations,[25] for which GO serves as a fluorophore and an elec-
tron donor whereas the metal ions act as the electron acceptor
(Figure 7b). GO is initially functionalized with a single-stranded
DNA (ssDNA) aptamer, which can capture Hg2 + and form a
hairpin structure on GO due to the formation of a thymine-
Hg2 + -thymine (T-Hg-T) complex. Once the GO-aptamer-Hg2 +
complex is formed, the fluorescence of GO is quenched due to
charge transfer from GO to Hg2 + . A similar sensor platform has
been used for I@ detection, as shown in Figure 7c.[27] Because
the binding constant[28] for Hg2 + and I@ is > 1029, much larger
than that for Hg2 + and thymine, which is about 106, HgI2 is
preferentially formed and pulls the Hg2 + ions away from the
GO surface, which leads to recovery of the quenched fluores-
cence and allows I@ to be detected. In addition, the GO sensor
platform based on charge transfer has been successfully ap-
plied for the detection of various other environmental and ag-
ricultural pollutants, such as Au3 + , Fe3 + , Fe2 + , Co2 + , Ni2 + , Cd2 +
, and HCl.[15d,e, 29]
GO as a donor in FRET
In a FRET process, GO can serve as an energy donor in which
its fluorescence gets quenched by an energy acceptor, such as
gold nanoparticles or organic dyes.[30] FRET is a nonradiative
energy-transfer process based on the dipole–dipole interac-
tion.[26] It requires the donor’s emission spectrum to overlap
with the acceptor’s absorption spectrum. The FRET efficiency is
inversely proportional to the sixth power of the separation dis-
tance between the donor and acceptor. Therefore, for FRET to
be efficient, the energy donor and the acceptor need to be
brought close to each other, usually within about 6 nm (the
Fçrster distance) if organic dyes serve as both the energy
donors and acceptors. This can be realized through, for exam-
ple, antigen–antibody interactions or DNA hybridization.[30b, 31]
Because GO has a broad fluorescence emission peak, the con-
dition of spectrum overlap can be easily met by using noble-
metal nanoparticles as the quencher because they have a large
absorption cross-section with absorption band tunable from
the UV to the NIR region. For example, GO has been used as
an energy donor in an immuno-biosensor system for rotavius
detection (Figure 8).[30b] Initially, gold nanoparticles were func-
tionalized with the detection antibody of the rotavirus, and
the capture antibody was immobilized the GO surface. The
gold nanoparticles and GO could be brought into proximity
through the antigen–antibody interaction after capture of the
rotavirus cell. Consequentially, the fluorescence of GO was
quenched by the gold nanoparticles by the FRET process. GO

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Figure 8. GO as a FRET donor in a sensor for rotavirus detection, in which
the fluorescence of GO is quenched by gold nanoparticles. Reprinted from
Ref. [30b] with permission from Wiley.
has also been exploited as an energy donor in FRET sensors for
drug detection, disease diagnosis, pharmaceutical screening,
and so on.[30a, 32]
GO as an acceptor in FRET
In a FRET process, GO can also act as an energy acceptor, in
which it quenches the fluorescence of an energy donor, such
as quantum dots and organic dyes.[29a, 33] The use of GO as a
quencher has several advantages: 1) The 2D planar surface
area renders a large number of bonding sites; 2) the oxygen-
containing functional groups enable further bioconjugation;
3) it is water-soluble, non-toxic, biocompatible, and able to
Figure 9. Schematic illustration of sensor design strategies with GO as a quencher. a) An aptamer-based sensor. b) A DNA-based sensor. c) A sandwich-struc-
tured immunoassay. d) A competitive immunoassay.
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Focus Review
penetrate into cells. Also, the effective quenching distance
could be extended to around 30 nm if GO is used as an
energy acceptor. Figure 9 highlights several FRET detection
schemes.[29a, 33] For example, 1) the fluorophore-labeled single-
stranded aptamers were initially captured on GO through the
p–p stacking force between the aromatic nucleobases of apta-
mer and the aromatic rings in GO, as shown in Figure 9a. The
fluorescence of the fluorophore, such as organic dyes and
quantum dots, was quenched by GO in proximity through the
FRET process. In the presence of analytes, the aptamers
formed hairpin or G-quadruplex structures, which have a weak
interacting force with GO, so they were released from GO and
turned on the fluorescence. 2) In Figure 9b, the fluorophore-la-
beled double-stranded DNA molecules were initially captured
on the GO surface, with one strand of DNA covalently bonded
to GO and the other strand of DNA labeled with a fluorophore.
Fluorescence of the fluorophore was quenched if the fluoro-
phore was controlled to be close to the GO surface (typically
less than 6 nm). The addition of analytes caused DNA dehy-
bridization, and the fluorophore-labeled DNA strand formed a
hairpin structure and detached from the GO surface. 3) In Fig-
ure 9c, the free-standing fluorophore-labeled detection anti-
bodies initially displayed fluorescence. When analytes were
present, the analytes were sandwiched between the detection
antibody and the capture antibody, which brought the fluoro-
phore close to the GO surface and led to quenching of the
fluorescence. 4) In Figure 9d, the fluorophore-labeled antigens
were initially attached to the capture antibody on the GO sur-
face, with the fluorescence quenched. After addition of ana-
lytes, the fluorophore-labeled antigens were displaced away
from GO due to the stronger affinity between the analyte and
the capture antibody, which led to recovery of fluorescence.
For example, microRNA (miRNA) is an important type of bio-
marker because it regulates the expression of diverse genes
and can be related to many diseases, such as cancers and dia-
betes. GO, with its ability to penetrate through intracellular
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Figure 10. GO as a quencher in sensors. a) miRNA detection based on nanosized graphene oxide (NGO) and peptide nucleic acid (PNA), in which the fluores-
cence of the dye label on the PNA probe was initially quenched by GO but later recovered after the probe detached from NGO and hybridized with a target
miRNA. Reprinted with permission from Ref. [33a]. Copyright (2013) American Chemical Society. b) Detection of the ovarian cancer biomarker CA-125, based
on CRET. Reproduced from Ref. [36] with permission of The Royal Society of Chemistry.

membranes, has been used to build FRET sensors for the in
vitro detection of miRNA, as shown in Figure 10a.[33a] In this
sensor, a dye-labeled peptide nucleic acid (PNA) was initially
quenched after being adsorbed onto GO. The GO-PNA-dye
complex was then delivered into the cytoplasm, in which the
target miRNA hybridized with the dye-labeled PNA and de-
tached from GO, which led to fluorescence recovery. In addi-
tion, GO as a FRET quencher has also been used for in vitro
and in vivo molecular probing based on two-photon excitation
spectroscopy.[34] By replacing the capture and detection re-
agents, the GO sensor platform has been extended for the de-
tection of proteins, DNA, drugs, heavy metals, and so on.[33b, 35]
In addition to fluorescence, chemiluminescence provides an
alternative for optical biosensing. Chemiluminescence occurs
as a result of chemical reactions rather than direct photoexcita-
tion, which could lead to a low signal-to-noise ratio. It has
been reported that the strong quenching capability of GO and
GQDs can be used to quench chemiluminescence through
chemiluminescence resonance energy transfer (CRET), as
shown in Figure 10b, in which the cancer biomarker CA-125
was detected based on CRET.[36] GQDs were used as the
quenchers (energy acceptors) and assembled on a glass sub-
strate, then further functionalized with the capture antibody
for CA-125. The detection antibody for CA-125 was labeled
with the horseradish peroxidase (HRP) enzyme. In the presence
of the CA-125 antigen, an antibody-antigen-antibody complex
was formed with HRP and brought in proximity to GQDs. As a
result, the HRP enzyme catalyzed H2O2 to produce reactive
oxygen species (ROS), which further oxidized luminol to gener-
ate chemiluminescence. If HRP is brought close to GQDs in the
presence of the CA-125 antigen, the produced chemilumines-
cence is quenched by nearby GQDs. This “turn-off” sensor ena-

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bled the CA-125 cancer biomarker to be detected sensitively.
In addition, CRET sensors with GO or GQDs as quenchers have
also been used for the detection of other protein biomarkers,
DNA, and environmental pollutants.[36–37]
GQDs in fluorescent biosensors
Although GQDs shares many common excellent properties
with GO, such as easy functionalization, photostability, water-
solubility, biocompatibility, and nontoxicity, GQDs have their
own unique properties. For example, GQDs have a much small-
er steric effect due to their small size and can easily penetrate
through intracellular membranes. This unique property makes
GQDs a candidate material in drug delivery and in vivo and in
vitro bioimaging.[38] For example, Lannazzo et al. employed
GQDs for cancer-targeted drug delivery.[38c] Because GQDs also
have an excitation-wavelength-dependent emission, it can dis-
play different colors in bioimaging under different excitations
to meet the needs of various types of biosensing, as shown in
Figure 11a.[38e] The ability of GQDs to penetrate through intra-
cellular membranes also allows in-depth bioimaging by using
NIR light, which enormously extends the scope of fluorescent
bioimaging, as shown in Figure 11b.[38d]
Conclusions and Perspectives
There are many superior properties of GO and GQDs. Com-
pared with organic dyes, GO and GQDs are photostable. Com-
Figure 11. Bioimaging with GQDs by using the giant red-edge effect. a) Fluorescence images of A549 cells cultured with GQDs under different excitations
(shown on the upper right of each image). Reprinted by permission from Macmillan Publishers Ltd: [Light: Science & Applications] (Ref. [38e]), copyright
(2015). b) Fluorescence images with GQDs at different depths in tissue. Reprinted with permission from Ref. [38d]. Copyright (2017) American Chemical Soci-
ety.
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pared with most inorganic semiconductor quantum dots, GO
and GQDs are nontoxic and biocompatible. The strong excita-
tion-wavelength-dependent fluorescence of GO and GQDs are
unique to fluorescent sensing and imaging, and can be ex-
tended to other photoluminescent devices. However, GO and
GQDs exhibit broad fluorescence peaks in the presence of a
large variety of oxygen-containing moieties, as compared with
the sharp fluorescence peak of organic dyes and inorganic
quantum dots. If a narrow fluorescence peak is needed for
some applications, it is necessary to purify and functionalize
GO.
Interestingly, GO is suitable for NIR biological imaging by
using two-photon excitation fluorescence spectroscopy due to
a large two-photon absorption cross-section. Conversely, GO
can be purified to eliminate various oxygen-containing moiet-
ies and further functionalized to tailor the fluorescence peak
position and width. In the future, further studies are needed to
tune the fluorescence of purified and functionalized GO.
GO and GQDs can be employed not only as fluorophores
but also as quenchers, which provides plenty of room for the
development of new fluorescent biosensors and bioimaging
systems. It is envisioned that the applications of GO and GQDs
in in vitro and in vivo sensing and imaging will continue to
expand.
Multiplex detection is significant for practical applications of
biosensors because it will enormously improve the specificity
and lower the cost of detection.[39] The dual roles of GO and
GQDs as a fluorophore and a quencher make them highly
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promising. They can transduce an excitation-wavelength-de-
pendent fluorescence signal and simultaneously quench the
fluorescence from external fluorophores, which has strong im-
plications in the detection of multiple events on a single bio-
sensor.
Furthermore, the 2D planar structure of GO makes it ideal to
couple with lab-on-chip devices.[40] The integration of all these
functionalities into a single chip would greatly promote point-
of-care and field-deployable applications of biosensing devices
in the future. It is expected that GO and GQDs, along with
other materials in the family of graphene-based materials, will
find increasing applications in healthcare, environmental pro-
tection, agriculture, food safety, homeland security, and so on.
Acknowledgements
Research reported in this publication was supported by the
National Institute of Neurological Disorders and Stroke of the
National Institutes of Health under award number
R15NS087515. The content is solely the responsibility of the
authors and does not necessarily represent the official views of
the National Institutes of Health. This work was also partially
supported by an NSF grant (CBET-1336205).
Conflict of interest
The authors declare no conflict of interest.
Keywords: biosensors · fluorescence · graphene · quantum
dots · sensors
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Manuscript received: June 1, 2017
Revised manuscript received: July 23, 2017
Accepted manuscript online: July 25, 2017
Version of record online: August 30, 2017
T 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim