Applied Surface Science 434 (2018) 155–162

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Applied Surface Science

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

Full Length Article

Graphene quantum dots modified with adenine for efficient

two-photon bioimaging and white light-activated antibacteria
Zhimin Luo a,1 , Dongliang Yang a,1 , Chen Yang a , Xiangyang Wu d , Yanling Hu a ,
Ying Zhang a , Lihui Yuwen a , Edwin Kok Lee Yeow d , Lixing Weng c , Wei Huang a,b ,
Lianhui Wang a,∗
a
Key Laboratory for Organic Electronics and Information Display (KLOEID) and Institute of Advanced Materials (IAM), Jiangsu National Synergetic
Innovation Center for Advanced Materials (SICAM), Nanjing University of Posts and Telecommunications, 9 Wenyuan Road, Nanjing 210023, China
b
Key Laboratory of Flexible Electronics (KLOFE) and Institute of Advanced Materials (IAM), Jiangsu National Synergistic Innovation Center for Advanced
Materials (SICAM), Nanjing Tech University (NanjingTech), 30 South Puzhu Road, Nanjing 211816, China
c
College of Geography and Biological Information, Nanjing University of Posts and Telecommunications, 9 Wenyuan Road, Nanjing 210023, China
d
Division of Physics and Applied Physics, School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore
637371, Singapore

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

Article history: A simple two-step microwave-assisted method was developed for the preparation of adenine-modified
Received 8 June 2017 graphene quantum dots (A-GQDs) with striking fluorescence properties. The prepared A-GQDs with the
Received in revised form 11 October 2017 lateral size of 3–5 nm and single- to few-layer thickness exhibit intense fluorescence with a quantum
Accepted 16 October 2017
yield of 21.63% (excited at 350 nm) and an average lifetime of 4.47 ns. The A-GQDs are biocompatible
Available online 18 October 2017
and present two-photon green fluorescence, which render them suitable for two-photon fluorescent
cell imaging. More interestingly, we demonstrate for the first time that the A-GQDs also possess white
Keywords:
light-activated antibacterial property. Due to their excellent photoluminescence performance and ease of
Graphene quantum dots
Adenine
biological conjugation, A-GQDs can be envisioned as an emerging role for many multifunctional biomed-
Fluorescence ical applications.
Two-photon bioimaging © 2017 Elsevier B.V. All rights reserved.
White light-activated antibacteria

1. Introduction
Graphene, an atomically thin carbon film with honeycomb
structure, attracts considerable attention due to its unique phys-
ical and chemical properties along with great potential for wide
ranges of applications such as electronics, sensors, energy stor-
age and conversion [1–6]. However, graphene’s zero bandgap and
lack of photoluminescence limit its application in biological fields
[7,8]. Towards this challenge, recent developments have focused
on graphene quantum dots (GQDs), which are composed of single-
to few-layer graphene with heteroatomic functional groups on the
surface. Unlike simple graphene, these GQDs have outstanding pho-
toluminescence properties due to quantum confinement and defect
effect on the surface. They exhibit many interesting characteristics
such as excitation wavelength-dependent fluorescent emission,
resistance to photobleaching, excellent biocompatibility, and ease
∗ Corresponding author.
E-mail address: iamlhwang@njupt.edu.cn (L. Wang).
1
These authors contributed equally to this work.
for bioconjugation [9–12]. Since traditional organic dyes are not
environmentally stable [13,14] whilst inorganic quantum dots (e.g.
CdS, CdTe, CdSe) usually contain poisonous heavy metal [15–18],
GQDs or their derivatives are the emerging alternatives of novel flu-
orescent nanomaterials to replace the conventional fluorophores.
However, current preparations of GQDs are usually based on the
chemical cutting of graphene oxide sheets [9,10]. These methods
suffer from complicated procedures and often result in weak fluo-
rescence of GQDs [19]. A more simple method for preparing highly
fluorescent GQDs or their derivatives is expected.
Singlet oxygen (1 O2 ) is a major active species to kill bacteria,
especially for exterminating Eschetichia coli (E. coli) via photo-
dynamic antibacterial chemotherapy (PACT) [20–23] in which
phorphyrin and its derivatives are often used as the photosensitizer
agents. For example, carbon nanotube/porphyrin composite [24]
and polythiophene/porphyrin complex [25] are able to produce
1 O to kill bacteria under white light illumination. However, these
2
materials show poor photostability due to photobleaching and
generally have no absorption in wavelengths longer than 600 nm
[26,27]. Even for recent GQDs finding in which 1 O2 was generated to

https://doi.org/10.1016/j.apsusc.2017.10.121
0169-4332/© 2017 Elsevier B.V. All rights reserved.
156 Z. Luo et al. / Applied Surface Science 434 (2018) 155–162
kill U251 human glioma cells, the experiment was carried out under
the irradiation of blue light (␭ = 470 nm) [28]. The capability to tune
the absorption wavelengths of GQDs for photodynamic antibacteria
comes handy in this case. We envision a novel antibacterial agent
that can be activated under white light and wide spectrum of light
(blue, green, red). Chemical modification of GQDs may be an effi-
cient way to enhance their photoluminescence [29,30], and even
improve their yield of singlet oxygen for antibacterial applications
[31]. Adenine, a nucleobase with a variety of roles in biochemistry,
contains amine groups, which was used for functionalizing carbon
nanotubes [32] or reduced graphene oxides [33] to extend their
applications.
Herein, we develop a facile two-step microwave-assisted
method to prepare adenine-modified GQDs (referred to as A-GQDs)
using bulk graphite and adenine as the raw materials (Scheme 1a).
The prepared A-GQDs exhibit high fluorescence with a quantum
yield of 21.63% at the excitation wavelength of 350 nm. Signifi-
cantly, the A-GQDs also present two-photon green fluorescence
when excited at 750 nm, which results in their good performance
in the two-photon fluorescent cell imaging. More intriguingly, for
the first time, we identify that the A-GQDs could act as highly effi-
cient photodynamic antibacterial agent for E. coli under white light
and wide spectrum of light (␭ = 450, 535, 635 nm).
2. Experimental section
2.1. Materials and reagents
Graphite (20 ␮m, Synthetic), adenine (Reagent grade, ≥99%),
propidium iodide (HPLC, ≥94%) and reactive oxygen species assay
kit were purchased from Sigma-Aldrich (Saint Louis, Missouri,
USA). Fetal bovine serum and Dulbecco’s modified Eagle’s medium
(DMEM) were purchased from Fisher Scientific (a part of Thermo
Fisher Scientific, Loughborough, UK). Milli-Q water (Milli-Q Sys-
tem, Millipore, Billerica, MA) was used in all the experiments. All
the reagents were used as received without further purification.
2.2. Preparation of GQDs
GQDs were prepared according to our previous reported method
[34]. 75 mg of graphite was ultrasonicated in 20 mL of mixed acid
(H2 SO4 :HNO3 = 3:1) for 2 h, and then the mixture was heated at
80 ◦ C for 4 h under microwave irradiation [34]. The resultant solu-
tion was filtered by microporous membrane (pore size 0.22 ␮m) to
get the GQDs solution containing residual acid. Sodium carbonate
was added into the GQDs solution to neutralize the residual acid.
The neutralized solution was dialyzed for 3 days to get the purified
GQDs aqueous solution, which was used as original material for
preparation of adenine modified GQDs.
2.3. Preparation of A-GQDs
20 mL of GQDs aqueous solution and 25 mg of adenine were
added into a quartz tube, and reacted in the microwave reactor
at 175 ◦ C for 12 h. The resultant colloidal solution was placed at
4 ◦ C for 12 h and then filtered by microporous membrane (pore
size 0.22 ␮m) to remove the residual adenine. The filtered solu-
tion was dialyzed for 2 days. The as-prepared colloidal solution was
evaporated and dried in vacuum drying oven over night to get the
A-GQDs.
2.4. Cytotoxicity assay
Human lung carcinoma A549 cells were used as a test model in
our experiments. The cells were incubated in DMEM supplemented
with 10% fetal bovine serum. Cell cultures were kept at 37 ◦ C in a
5% CO2 humidified incubator and harvested through centrifugation
at 750 r.p.m. for 3 min. The viability of cell was evaluated with MTT
method. The cells (1 × 104 ) were seeded on each well of 96 multi-
well plates, and then were incubated with different concentrations
of A-GQDs in DMEM (2, 1, 0.1, 0.01, 0.001, 0.0001 mg mL−1 ) at 37 ◦ C
for 24 h. After incubation, the culture medium was removed and
the cells were washed with 10 mM phosphate-buffered saline (PBS)
solution (pH = 7.0) for three times. Subsequently, 90 ␮L of fresh cell
culture medium and 10 ␮L of MTT solution (5 mg mL−1 ) were suc-
cessively added into each well. The cells were incubated for 4 h.
After the formanzan dye produced by live cells was dissolved with
DMSO (150 ␮L), the absorbance of cell culture solution at 490 nm
was recorded using a Microplate Spectrophotometer (PowerWave
XS2, BIOTEK, US). Cell viability was evaluated with the percentage
of the absorbance of cells incubated with A-GQDs to that of cells
cultured in normal culture medium.
2.5. Cell imaging
Twenty microliters of A-GQDs suspension solution
(10.9 mg mL−1 ) was added into 2 mL of DMEM containing human
lung carcinoma cell A549 in 35 mm plate (the final concentration
of A-GQDs in DMEM is 0.109 mg mL−1 ). The plate was incubated
at 37 ◦ C for 21 h and then the medium was removed. The cells
were washed with 10 mM PBS solution (pH = 7.0) for two times
before being taken an image. The fluorescent images for human
lung carcinoma cell A549 were recorded on confocal microscope
(Olympus, FV1000, Japan) at the excitation of 405 and 750 nm.
2.6. White light-activated antibacterial activity of A-GQDs
A single colony of E. coli DH5␣ (Gram-negative) on a solid Luria-
Broth (LB) agar plate was transferred to 5 mL of liquid LB culture
medium and incubated at 37 ◦ C for 12 h under aerobic condition.
Bacteria were harvested by centrifugation at 7000 r.p.m. for 3 min.
The obtained bacterial cells were washed thoroughly by 10 mM PBS
solution (pH = 7.0) for three times. The supernatant was carefully
removed through centrifugation, and the remaining bacteria were
resuspended in the 10 mM PBS solution (pH = 7.0). The bacterial
suspension was adjusted to be 0.1 optical density (OD) at 600 nm
measured by UV–vis spectrometer, which was referred as bacte-
rial inoculums for use in the next step. 8 mL of A-GQDs solution
(10.9 mg mL−1 ) was filtrated by microporous filter (0.22 ␮m), and
mixed with 3 mL of LB broth to form the A-GQDs complex. 200 ␮L
of A-GQDs complex impregnated with 8 ␮L of the above prepared
bacterial inoculums, were deposited on sterile six-well plates, and
incubated at 37 ◦ C for 21 h under the irradiation of white light in wet
atmosphere. The bacterial suspensions were sufficiently mixed, and
carefully diluted 2 × 105 fold with 10 mM PBS solution (pH = 7.0).
50 ␮L of diluted samples were then spread on LB agar plates. After
being incubated at 37 ◦ C for 21 h, plates were counted to determine
total colony-forming units (CFU) per mL of undiluted suspension.
Each test was done in triplicate and conducted along with neces-
sary controls, such as a LB broth with A-GQDs was incubated at
37 ◦ C for 21 h in the dark, and a LB broth without A-GQDs were
incubated at 37 ◦ C for 21 h in the dark or under the irradiation of
white light. S. aureus ATCC 25923 was used as a model for testing
the white light-activated antibacterial activity of A-GQDs to Gram-
positive bacteria. The experimental method is the same as that of
E. coli DH5␣.
2.7. Cell death analysis
E. coli was dispersed in 10 mg mL−1 A-GQDs solution and irra-
diated with white light (2.35 mW) for 12 h. The E. coli cells were
then incubated with 10 ␮g mL−1 propidium iodide for 15 min in the
 Z. Luo et al. / Applied Surface Science 434 (2018) 155–162
Scheme 1. (a) Schematic depicting the microwave-assisted preparation of A-GQDs. (b) white light-activated antibacterial property of A-GQDs towards E. coli.
dark. Propidium iodide-stained death cells images were recorded
by a fluorescent microscope (IX71, Olympus, Japan) at the excita-
tion wavelength of 559 nm. Experiments for antibacteria activated
by blue, green, red light were also carried out by the irradiation
with 450, 535 and 635 nm light, respectively.
3. Results and discussion
3.1. Preparation and characterizations of A-GQDs
In our experiments, fluorescent GQDs were firstly prepared
by microwave-assisted exfoliation and cutting of graphite in the
mixed strong acid solution (H2 SO4 :HNO3 = 3:1), and then GQDs
were chemically modified with adenine under microwave irradia-
tion to synthesize A-GQDs with high fluorescence. The prepared
GQDs and A-GQDs were characterized by transmission electron
microscopy (TEM), X-ray photoelectron spectroscopy (XPS), Raman
spectroscopy, and Fourier transform infrared spectroscopy (FTIR).
The TEM image (Fig. S1) and AFM analysis (Fig. S2) show that the lat-
eral size and height distribution of GQDs is 3–5 nm and 0.75–3.8 nm,
respectively, indicating that the GQDs is single- to few-layer thick-
ness. As shown in Fig. 1a, the high-resolution TEM (HRTEM) image
of A-GQDs confirms their lateral size is still less than 5 nm even
after the chemical modification of GQDs with adenine. The HRTEM
image (Fig. 1b) indicates that A-GQDs are highly crystalline with
crystal lattice spacing of 0.209 nm, which can be ascribed to the
(100) plane of the graphitic structure [35,36].
The composition of A-GQDs was confirmed by the XPS mea-
surements in which the spectra (Fig. 2a) show a graphitic C1s
peak at 284.4 eV and an O1s peak at 531 eV [36]. An obvious
N1s peak at 399.7 eV was also observed, indicating a successful
adenine modification on GQDs [12,37]. The high-resolution C1s
spectrum (Fig. 2b) exhibits the C C (284.4 eV), C C (285.4 eV), C N
(286.2 eV), and C O (287.6 eV), indicating that A-GQDs contain the
oxygenated and nitrogen-functionalized carbon [12,37]. The main
157
binding energy at 399.7 eV in the high-resolution N1s spectrum of
A-GQDs (Fig. 2c) is ascribed to the amide ( CO NH ), which fur-
ther confirms the chemical reaction between GQDs and adenine
under the microwave irradiation [38,39].
The typical D band (1340 cm−1 ) and G band (1545 cm−1 ) were
detected in the Raman spectrum of the A-GQDs (Fig. 2d) [40].
The intensity ratio of D/G is 1.04, which is much higher than
that of the pristine graphene, indicating that the graphitic struc-
ture of the A-GQDs was partially destroyed during the chemical
modification. The chemical modification of GQDs with adenine
was confirmed by the FTIR spectra (Fig. S3). Fig. S3a shows sev-
eral distinct absorption peaks of GQDs: 1) 1722 cm−1 for the C O
stretching vibration of COOH; 2) 1612 cm−1 for the O H bend-
ing vibration and graphene skeletal ring vibration; 3) 1130 cm−1
for aromatic C H in-plane deformation vibration [41–43]. Lack of
absorption peak at 1722 cm−1 for COOH and an absorption peak
at 1633 cm−1 ascribed to amide I band in the FTIR spectrum of A-
GQDs indicate the formation of CONH by the chemical reaction
between GQDs and adenine [44].
The optical properties of GQDs and A-GQDs were investigated by
UV–vis and fluorescence spectrophotometer. The absorption spec-
trum of GQDs has an evident peak at 270 nm and a weak peak at
310 nm (Fig. S4a). As shown in Fig. S5a, after modification with
adenine, the absorption peak at 270 nm redshifts to 312 nm. GQDs
emit weak blue-green light at the excitation of 350 nm (Fig. S4b).
However, a strong emission peak at 420 nm (Fig. 3a) was observed
when the A-GQD aqueous solution was excited at 350 nm. The
corresponding bright blue fluorescence of A-GQDs under the ultra-
violet irradiation of 350 nm can be seen in the photograph (inset
in Fig. 3a). The fluorescent quantum yield of the A-GQD was mea-
sured to be 21.63% by using quinine sulfate as a reference (Table
S1), which is higher than that of those reported fluorescent GQDs
[9,10,12,37,38]. When A-GQDs aqueous solution was excited at the
750 nm, it displays evident two-photon green fluorescence (Fig. 3b).
It is also found that the emission spectra of A-GQDs are depen-
158 Z. Luo et al. / Applied Surface Science 434 (2018) 155–162

Fig. 1. (a) TEM and (b) HRTEM images of A-GQDs with inset in (b) representing the crystal lattice of A-GQD.

Fig. 2. (a) Survey XPS spectrum, high-resolution (b) C1s and (c) N1s XPS spectra of A-GQDs. (d) Raman spectrum of A-GQDs.

dent on excitation wavelengths. As shown in Fig. S5b, the emission longer than that of GQDs prepared via amidative cutting of tattered
peak shifts to the longer wavelength as the excitation wavelength graphite [12] or derived from carbon fibers [37].
changes from 325 to 450 nm. These excitation-dependent emis-
sion behaviors are often observed for fluorescent carbon-based
3.2. Cytotoxicity of A-GQDs
nanomaterials and here it may result from optical transitions of
A-GQDs with different sizes and surface defects [9,10]. The fluo-
Due to its high fluorescence quantum yield and excitation-
rescence decay profile of A-GQDs was measured at the excitation
dependent fluorescent character, A-GQDs are suitable candidate for
wavelength of 375 nm by a time-correlated single photon counting
applications in bioimaging. Therefore, we further examined their
technique (Fig. 3c). The lifetime of A-GQDs is well fitted to a triple-
cytotoxicity using human lung carcinoma A549 cell as a test model
exponential function and measured as ␶1 = 0.48 ns (B1 = 8.68%),
of MTT viability assay. It can be observed from Fig. 3d that A-GQDs
␶2 = 2.7 ns (B2 = 39.55%), and ␶3 = 6.5 ns (B3 = 51.76%) [37,45,46]. The
have good biocompatibility and show excellent cell viability at the
average fluorescent lifetime is calculated to be 4.47 ns accord-
concentration of less than 2 mg mL−1 . The cell viability can still keep
ing to the equation of ␶ = B1 ␶1 + B2 ␶2 + B3 ␶3 [37,45,46], which is
94%, even if the human lung carcinoma A549 cells were incubated
with A-GQDs (2 mg mL−1 ) for 24 h.
 Z. Luo et al. / Applied Surface Science 434 (2018) 155–162
Fig. 3. Fluorescent emission spectra of A-GQDs at the excitation wavelengths of (a) 350 nm and (b) 750 nm. Inset in (a) showing the image of A-GQDs colloidal solution when
exposed to the ultraviolet irradiation (␭ = 350 nm). (c) Fluorescent lifetime of A-GQDs at the excitation wavelength of 375 nm. (d) Viability of human lung carcinoma A549
cell incubated with different concentrations of A-GQDs for 24 h.
3.3. Fluorescent bioimaging and photo-activated antibacteria of
A-GQDs
Cell imaging is an advanced biological technique for medical
diagnosis. For instance, the early detection of lung cancer and its
subsequent prevention can be carried out through cell imaging due
to the deformed shapes of human lung cell’s nuclei in the procedure
of lung cancer. A-GQDs were applied for cell imaging using human
lung carcinoma A549 cell as a model. The cells were incubated in
Dulbecco’s modified Eagle’s medium (DMEM) containing A-GQDs
at 37 ◦ C for confocal fluorescent imaging. The phase contrast image
and confocal fluorescent images of human lung carcinoma A549
cell at the excitation wavelengths of 405 and 750 nm were shown
in Fig. 4. A blue fluorescent cell image can be observed at the excita-
tion wavelength of 405 nm. Fluorescent A-GQDs mainly distribute
around each cell nucleus, indicating an efficient A-GQDs internal-
ization to the human lung carcinoma A549 cell. The two-photon
green fluorescent image at the excitation wavelength of 750 nm is
clearer due to less fluorescence interference from the background
in cells at this wavelength. These results of cell imaging suggest that
A-GQDs can be used as fluorescent label for high-contrast fluores-
cent bioimaging, especially in two-photon fluorescent technique.
The white light-activated antibacterial activity of A-GQDs
against Gram-positive Staphylococcus aureus (S. aureus) and Gram-
negative E. coli were also explored. Cell wall of Gram-positive
bacteria consists of polysaccharide and polypeptide chains, which
are positively charged and can prevent A-GQDs internalization into
cell through their electrostatic repulsion with A-GQDs (zeta poten-
tial 2.57 mV at pH 7.0). Therefore, the A-GQDs are not able to
have an effect on Gram-positive S. aureus under the irradiation of
white light (Fig. S6 and Fig. S7). The results of white light-activated
159
antimicrobial activity towards Gram-negative E. coli are shown in
Fig. S8. E. coli can grow normally in the dark (Fig. S8a) or under
the irradiation of white light (Fig. S8b) without A-GQDs whilst
only normal growth in the dark was observed after addition of
A-GQDs on the LB agar plate (Fig. S8c). However, there is almost
no growth of E. coli under the irradiation of white light on the
LB agar plate containing A-GQDs (Fig. S8d). The E. coli count (Fig.
S9) corresponding to antibacterial experiments of A-GQDs (Fig. S8)
confirms their excellent white light-activated antibacterial activity
against E. coli. The antibacterial experiment was also carried out
by propidium iodide (PI) stain (Fig. 5), which further proves that
A-GQDs have good antibacterial activity towards E. coli under irra-
diation of white light. Since white light contains wide ranges of
wavelengths (blue, green, red), the effect of each light on the pho-
toactivated antibacterial activity of A-GQDs was also explored. As
shown in Fig. 6, blue (450 nm), green (535 nm) and red (635 nm)
light all have an effect on the photo-activated antibacterial activity
of A-GQDs, indicating the broad-spectrum antibacterial activity of
A-GQDs. Until now, no report on the carbon-based nanomaterials
has shown white light-activated antimicrobial activity. Preceding
studies only demonstrated that antibacterial activity from carbon-
based nanomaterials, such as graphene oxide (GO) and reduced
graphene oxide sheets (rGO), arises from the effect of physical dis-
ruption of GO or rGO [47]. After GO or rGO was cut into GQDs,
their small size deactivates their antibacterial effect due to the loss
of physical disruption. In our experiment, similar behavior can be
confirmed by observing the lack of antibacterial activity for A-GQDs
in the dark. On the other hand, GQDs have been used as a novel
photodynamic therapy agent because they can produce 1 O2 via a
multistate sensitization process [36]. Therefore, the special surface
structure and ability of producing 1 O2 may contribute to their white
160 Z. Luo et al. / Applied Surface Science 434 (2018) 155–162

Fig. 4. (a) Phase contrast image of human lung carcinoma cell A549. Fluorescent images of human lung carcinoma cell A549 at the excitation wavelengths of (b) 405 nm and
(c) 750 nm.

Fig. 5. Optical images of E. coli incubated without A-GQDs (a) in the dark and (b) under the irradiation of white light. Optical images of E. coli incubated with A-GQDs (c)
in the dark and (d) under the irradiation of white light. Fluorescent images of E. coli incubated without A-GQDs (e) in the dark and (f) under the irradiation of white light.
Fluorescent images of E. coli incubated with A-GQDs (g) in the dark and (h) under the irradiation of white light. E. coli cells were stained with propidium iodide before imaging
by fluorescent microscope.

Fig. 6. Optical images of E. coli incubated with A-GQDs (a) in the dark and (b) under the irradiation of 450 nm light. Fluorescent images of E. coli incubated with A-GQDs
(g) in the dark and (h) under the irradiation of 450 nm light. Optical images of E. coli incubated with A-GQDs (c) in the dark and (d) under the irradiation of 535 nm light.
Fluorescent images of E. coli incubated without A-GQDs (i) in the dark and (j) under the irradiation of 535 nm light. Optical images of E. coli incubated with A-GQDs (e) in the
dark and (f) under the irradiation of 635 nm light. Fluorescent images of E. coli incubated without A-GQDs (k) in the dark and (l) under the irradiation of 635 nm light. E. coli
cells were stained with propidium iodide for imaging by fluorescent microscope.

light-activated antibacterial activity against E. coli. A-GQDs are pos- and made the E. coli. biocidal since 1 O2 was well known for killing
itively charged and can be strongly interacted with the bacterial bacteria [27]. The mechanism of white light-activated antibacterial
membrane of E. coli cell for internalization. When the A-GQDs were activity of A-GQDs was illustrated in the Scheme 1b, which was fur-
exposed to white light, 1 O2 was produced to destroy cell membrane ther proved by the control experiments that more reactive oxygen
 Z. Luo et al. / Applied Surface Science 434 (2018) 155–162
species were generated in the presence of A-GQDs under the irra-
diation of white light than that in the dark or without A-GQDs (Fig.
S10a). The yield of reactive oxygen species under the irradiation
of light with different wavelengths of 450, 535, 635 nm (Fig. S10b)
further confirms the origin of white light-activated antibacterial
activity of A-GQDs.
4. Conclusions
In summary, adenine modified GQDs, i.e. A-GQDs, with size
ranges from 3 to 5 nm were prepared via a two-step microwave-
assisted method from graphite and adenine precursors. The
resulting A-GQDs display high fluorescence with a quantum yield
efficiency of 21.63% and two-photon green fluorescence. A-GQDs
also show good biocompatibility towards human lung carcinoma
A549 cell. It makes them appealing for high-contrast fluorescent
cell imaging. More significantly, for the first time, A-GQDs are also
identified to be highly efficient white light-activated antibacterial
reagent, opening novel biological applications for GQDs.
Acknowledgements
This work was financially supported by the Ministry of Science
and Technology of China (2017YFA0205302), the Sci-tech Sup-
port Plan of Jiangsu Province (BE2014719), the Jiangsu National
Synergistic Innovation Center for Advanced Materials (SICAM),
the Priority Academic Program Development of Jiangsu Higher
Education Institutions (PAPD, YX03001), National Natural Sci-
ence Foundation of China (61705113) and the University Science
Research Project of Jiangsu Province (NY217007).
Appendix A. Supplementary data
Supplementary data associated with this article can be found,
in the online version, at https://doi.org/10.1016/j.apsusc.2017.10.
121.
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