Chemical Engineering Journal 431 (2022) 134110

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Chemical Engineering Journal

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

Two-photon photoluminescence and bio-imaging application of

monodispersed perovskite-in-silica nanocrystals with high biocompatibility
Chu-Yao Zhong a, Liangyan Xiao b, Jielin Zhou c, Zhifeng Chen c, Yibo Chen a, *, Zhao-Qing Liu a,
Jin Z. Zhang d
a
School of Chemistry and Chemical Engineering/Guangzhou Key Laboratory for Clean Energy and Materials, Guangzhou University, Guangzhou 510006, China
b
Guangdong Provincial Key Laboratory of Nanophotonic Functional Materials and Devices, School of Information and Optoelectronic Science and Engineering, South
China Normal University, Guangzhou 510006, China
c
School of Physics and Materials Science, Guangzhou University, Guangzhou 510006, China
d
Department of Chemistry and Biochemistry, University of California, Santa Cruz, CA 95064, USA

A R T I C L E I N F O A B S T R A C T

Keywords: Developing two-photon photoluminescence bio-probes with high-biocompatibility is still a challenge. In this
Two-photon photoluminescence work, single CsPbBr3 perovskite NCs were encapsulated into a silica nanoshell to prevent leakage of Pb2+ ion out
Bio-probes of the shell, and their two- and one-photon photoluminescence property, as well as the application potential for
Perovskite-in-silica Nanocrystals
bio-imaging, were investigated by detailed optical characterization and two-photon cell imaging experiment. It
turns out that the core–shell NCs show good two-photon absorption property under the excitation of an 800 nm
femtosecond laser and emit green light peaked at 526 nm. The survival rate of human hepatocellular carcinomas
(HepG2) cells maintains 96.21% after incubating with the core–shell NCs (16 μg/mL) for 24 h without adding
protective ligands, demonstrating the excellent biocompatibility of the nanoprobes. Based on the excellent
biocompatibility and good two-photon absorption property of the perovskite-in-silica nanoprobes, fluorescence
imaging of HepG2 cells was demonstrated under 800 nm femtosecond laser illumination. The cumulative fluo­
rescence intensity of the two-photon fluorescence images was five times higher than the control group. This work
demonstrates the good two-photon photoluminescence property of well-controlled core–shell perovskite-in-silica
NCs and provides a new insight for the promising applications in bio-imaging.

1. Introduction All-inorganic lead perovskite nanocrystals (CsPbX3 NCs, X = Cl, Br,

I), as a star material in the field of semiconductor optoelectronics
Early detection and diagnosis are effective ways to improve the [12–15], have shown large nonlinear absorption and low-threshold
curing rate of cancer, which is of great significance for human health. multiphoton pumped amplified spontaneous emission (ASE) [16–19].
NIR laser can be focused in a very small domain and offers advantages However, it is rarely been reported as a two-photon fluorescence bio­
such as high spatial resolution, deep penetration, low background logical probe due to the intrinsic instability and the toxicity of Pb2+ ions.
fluorescence, and little photodamage to tissues [1–3]. Thus, two-photon Silica (SiO2) coating is an effective strategy for protecting colloidal
fluorescence imaging, which utilizes two near-infrared (NIR) photons as nanoparticles from reactive species such as moisture, improving their
the excitation source and results in the emission of visible light, is photo-stabilities, and reducing the diffusion of toxic ions [20–26]. For
considered a powerful tool to monitor the dynamics of cells and detect example, Hu et al. embedded CsPbBr3 quantum dots (QDs) into silica
early pathological changes [4–7]. In recent years, biological imaging shells to suppress blinking while obtaining improved moisture resistance
under NIR light utilizing lanthanide-doped nanoparticles, aggregation- and enhanced photostability [22]. However, the size of the micrometer-
induced emission nanoparticles, and II-VI quantum dots have aroused scale structures is too large for biomedical applications. Song et al. used
extensive attention [3,8–11]. Even though, it is of strong interest to a “stress-response” method to encapsulate CsPbBr3 NCs into silica nano-
develop high-efficiency, high-biocompatibility, and low-cost two- shells (~3.5 nm) and used them as fluorescence probes for tumor cell
photon fluorescence probes. imaging [23]. Unfortunately, they conducted the cell imaging under UV

* Corresponding author.
E-mail address: chenyibo@gzhu.edu.cn (Y. Chen).

https://doi.org/10.1016/j.cej.2021.134110
Received 26 August 2021; Received in revised form 12 November 2021; Accepted 8 December 2021
Available online 14 December 2021
1385-8947/© 2021 Elsevier B.V. All rights reserved.
C.-Y. Zhong et al. Chemical Engineering Journal 431 (2022) 134110

illumination, which has low penetration depth in biological tissues, and A1, Zeiss, Berlin, Germany), and a spectrometer (SR-500i-B1, Andor,
the related application is limited to surface cells. Oxford, UK). The two-photon PL lifetime was measured using an 800 nm
In this work, we synthesized monodispersed lead bromide femtosecond (fs) laser, detected by an amplified fast detector, and
perovskite-in-silica core–shell NCs in a facile one-step heterogeneous measured by an oscilloscope. The laser pulse duration was ~ 100 fs, and
nucleation process and continuously controlled the silica shell thickness the excitation fluence was ~ 17 J/cm2. The temporal resolution of the
in the range of 15 ~ 60 nm. All the as-prepared samples show strong system is ~ 2 ns. After immersing the NCs in DI water for 24 h with the
two-photon absorption, and the photoluminescence (PL) is dependent initial concentration of 50 μg/mL, the concentration of leaked Pb2+ in
on the silica shell thickness. The lead bromide perovskite-in-silica cor­ the supernatant was measured by an inductively coupled plasma mass
e–shell NCs also show good biocompatibility and strong performance in spectrometer (iCAP Qc ICPMS, Thermo Fisher Scientific).
two-photon fluorescence imaging of HepG2 cells under an 800 nm
pulsed laser. This study demonstrates the potential of core–shell 2.5. Cell culture
perovskite-in-silica NCs in two-photon fluorescence bio-imaging.
The human hepatocellular carcinomas (HepG2) cells used in this
2. Experimental section experiment were purchased from the Cell Lab of the Cell Resource
Center of the Chinese Academy of Sciences. The cells were seeded into
2.1. Raw materials the culture medium at 105 cells/dish and incubated in a humidified
incubator at 37 ℃ with 5% CO2. The culture medium is consisting of
Lead bromide (PbBr2, 99%, Aladdin), cesium bromide (CsBr, 99.9%, 89% Dulbecco’s Modified Eagle Medium (DMEM), 10% fetal bovine
Aladdin), oleylamine (OAm, 80%-90%, Macklin), oleic acid (OA, AR, serum, and 1% penicillin/streptomycin. To keep the cells active, the
Aladdin), tetramethoxysilane (TMOS, 98%, Aladdin), N, N-Dime­ culture medium was changed every three days.
thylformamide (DMF, 99.9%, Macklin), ammonia solution (25%,
Guangzhou Chemical Reagent Factory), toluene (HPLC, Guangzhou 2.6. Cell imaging
Chemical Reagent Factory) were used directly without further
purification. The cultured HepG2 cells were washed with phosphate buffer saline
(PBS) solution and then cultured in a new medium containing 16 μg/mL
2.2. Synthesis of lead bromide perovskite-in-silica nanocrystals (NCs) CPB@SiO2-100 NCs as well as the above ingredients for 24 h at 37 ℃
with 5% CO2. During this process, the NCs entered the cell through
The lead bromide perovskite-in-silica NCs were prepared according endocytosis. After that, the cells were washed with PBS solution again,
to a previous report with modifications [24]. In a typical procedure, and a small amount of PBS solution was further added to keep the cells
0.40 mmol PbBr2, 0.30 mmol CsBr, 0.6 mL OAm and 2.0 mL OA were moist. Finally, the two-photon fluorescence images of HepG2 cells were
added in 10 mL DMF. The mixture was stirred in an oil bath at 90 ℃ for obtained using laser scanning microscopy (Nikon A1, Japan) under the
2 h, and a clear precursor solution was obtained. After that, 40 μL of excitation of an 800 nm femtosecond laser.
2.8% ammonia solution was added in 2 mL of the above precursor so­
lution. Then the mixed solution was added into 100 mL toluene con­ 3. Results and discussions
taining 100 μL TMOS under vigorous stirring and maintained for 120
min. The final product was obtained by centrifugation and washed three In this work, we successfully constructed monodispersed perovskite-
times with toluene. The products obtained by adding 75 μL TMOS (25 μL in-silica nanocrystals for two-photon bio-imaging (Fig. 1a). Firstly,
ammonia), 100 μL TMOS (30 μL ammonia), 125 μL TMOS (35 μL CPB@SiO2 core–shell NCs with different SiO2 thicknesses were synthe­
ammonia), and 150 μL TMOS (40 μL ammonia) were labeled as sized by changing the amount of TMOS (Fig. 1b), and the corresponding
CPB@SiO2-75, CPB@SiO2-100, CPB@SiO2-125, and CPB@SiO2-150, TEM images are shown in Fig. 1c-f. With an increasing amount of TMOS,
respectively. the thickness of the SiO2 shell gradually increases from ~ 15 to ~ 60 nm,
while the core diameter remains around ~ 25 nm. The formation of SiO2
2.3. Synthesis of lead bromide perovskite nanocrystals (NCs) shell can be characterized by Fourier transform infrared spectroscopy
(FTIR), as shown in Fig. 1g. The sharp peaks at 1090, 796, and 465 cm− 1
2 mL of the above precursor solution was quickly injected into 100 are due to the symmetric stretching vibrations of Si-O-Si and Si-O,
mL toluene, stirred vigorously for 10 min to obtain a yellow-green so­ indicating the existence of silica. The asymmetric and symmetric
lution. CsPbBr3 NCs powder was obtained by centrifugation and washed stretching vibrations of the CH2 at 2923 and 2854 cm− 1, as well as the
with toluene. bending vibrations of the NH2 bond at 1560 cm− 1, can be designated to
oleic acid and oleylamine as the surface ligands [23,24,27]. The ab­
2.4. Physicochemical characterizations sorption at 2365 cm− 1 is attributed to the C = O stretching vibration
caused by the sample absorbing CO2 from the atmosphere [28–29]. In
Transmission electron microscope (TEM, JEM-2100F) with Energy the Raman spectra of CPB@SiO2 NCs at room temperature (Fig. 1h), the
Dispersive X-ray Spectrometer (EDS) was used to analyze the surface band at 50 ~ 100 cm− 1 are reported as mostly related to [PbBr6]4-, and
morphology and crystal microstructure of the NCs. X-ray diffraction the broadband at 110 ~ 150 cm− 1 are mostly from Cs+ ions [30,31].
(XRD, PANalytical PW3040/60) with Cu-Kα radiation (λ = 0.15418 nm) Both of them shift to lower frequency with the growing size of the
was used to characterize the phase composition of the NCs. X-ray CPB@SiO2 NCs, which can be attributed mainly to the heavier silica
photoelectron spectroscopy (XPS, Thermo K-Alpha) was used to inves­ shells. The above results support that CsPbBr3 NCs were successfully
tigate the compositions and valence states of the NCs. A fluorescence encapsulated in silica shells, forming core–shell structures with
spectrometer (Edinburgh, FLS 1000) was used to measure the photo­ controllable shell thickness. In addition, to evaluate the effect of SiO2 on
luminescence (PL) spectra, PLQY spectra, and fluorescence lifetime preventing leaking of Pb2+, the concentration of Pb2+ in the supernatant
under 365 nm excitation. On the Hitachi UV-3010 spectrophotometer, solution after the NCs were immersed in DI water for 24 h was measured
BaSO4 was used as a reference to detect the UV–vis absorption spectra of by inductively coupled plasma mass spectrometer (ICP-MS). The values
the NCs. The Raman spectra were recorded using a Raman spectrometer for CsPbBr3, CPB@SiO2-75, CPB@SiO2-100, CPB@SiO2-125, and
(Thermo Fisher Scientific) with a 785 nm laser. The two-photon fluo­ CPB@SiO2-150 NCs were 20.14, 8.45, 6.86, 6.42, and 5.69 ppm,
rescence spectra were obtained using a mode-locked Ti: sapphire oscil­ respectively (Fig. 1i and Table S1). Obviously, as the thickness of the
lator (Mira 900S, Coherent, US), an inverted microscope (Axio Observer silica shell increases, the concentration of leaked Pb2+ becomes smaller

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Fig. 1. (a) Schematic diagram of CPB@SiO2 core–shell NCs for two-photon bio-imaging. (b) Relationship between amount of TMOS and size of core and shell. TEM
images of (c) CPB@SiO2-75, (d) CPB@SiO2-100, (e) CPB@SiO2-125, and (f) CPB@SiO2-150 NCs; the scale bar is 50 nm. (g) FTIR spectra of CPB@SiO2 NCs. (h)
Raman spectra of CPB@SiO2 NCs with different TMOS adding amount at room temperature. (i) Concentration of Pb2+ in supernatant solution.

and smaller, and the concentration of leaked Pb2+ ions in CPB@SiO2 and Cs 3d5/2 are centered at 738.2 and 724.3 eV while the peaks of Pb
NCs is significantly lower than that of pure CsPbBr3 NCs. It indicates that 4f5/2 and Pb 4f 7/2 are centered at 142.7 and 137.8 eV, which demon­
SiO2 can effectively prevent the loss of Pb2+ and that CPB@SiO2 NCs strate the existence of Cs(I) and Pb(II), respectively [25,32,33].
have lower cytotoxicity, which suggested that CPB@SiO2 NCs have the To determine how the TMOS concentration affects PL, a series of
possibility of bio-imaging. UV–vis (Fig. 3a) and PL spectra (Fig. 3b) at room temperature were
As a typical sample, the microstructure of CPB@SiO2 − 100 NCs was measured and compared. The thickness of the silica shell has no sig­
characterized. The energy-dispersive X-ray spectrometry (EDS) mapping nificant effect on the absorption of the NCs, and the UV–vis absorption
of CsPbBr3@SiO2-100 NCs shown in Fig. 2a reveals a monodisperse onset is ~ 505 nm. In addition, the PL emission peak of CPB@SiO2 NCs
core–shell structure with a core size of about 25 nm and a shell thickness is ~ 517 nm, which can emit bright green light under UV excitation
of about 30 nm, in which Br, Cs, and Pb are distributed in the central (Fig. 3c). As shown in Fig. 3d, the peak position and FWHM of PL spectra
part, while Si and O are deposited around. In the EDS linear scanning hardly change, indicating that the CPB NCs with different silica shell
spectrum (Fig. 2b), the energy peaks of Br, Cs, and Pb are weak at the thicknesses undergo no phase or band structure change. However, based
edges and strong at the center, while the energy peaks of Si and O are on the PL spectra (Fig. 3b), the amount of TMOS added strongly affects
strong at the edges and weak at the center, demonstrating the core–shell the PL intensity of the CPB@SiO2. With 100 μL of TMOS (the SiO2 shell
structure. The XRD patterns of the prepared samples (Fig. 2c and Fig. S1) thickness is ~ 30 nm), the PL intensity is the highest (Fig. 3f). This seems
are consistent with the standard ones of orthorhombic CsPbBr3 to suggest that optimized TMOS amount can generate a silica shell that
(ICSD#01–072-7929). High-resolution transmission electron micro­ completely covers the CsPbBr3 surface to alleviate aggregation
scopy (HRTEM) image (Fig. S2) shows a distinct lattice spacing of 0.42 compared to lower TMOS dosage. Too much TMOS seems to result in
nm for the CPB@SiO2-100 NCs, corresponding to the (0 2 0) crystal solid silica spheres (Fig. S4) and leads to less effective passivation and
planes of orthorhombic CsPbBr3. The X-ray photoelectron spectroscopy reduced PL intensity. The photoluminescence quantum yields (PLQY)
(XPS) of the CPB@SiO2 NCs are shown in Fig. 2d-h and Fig. S3. The main were also measured and compared. With the increased amount of TMOS,
peak of the O 1 s spectrum at 532.9 eV and Si 2p band peaked at 102.8 the variation trend of PLQY (Fig. S5) is consistent with the intensity of
eV confirm the formation of the SiO2 shell. The shoulder peak of O 1 s PL. What’s more, the PLQY value of the CPB@SiO2-100 NCs is more than
spectrum at 531.1 eV is attributed to –COOH of the residual oleic acid on two folds of the original CsPbBr3 NCs without the silica shell. Time-
the surface. In addition, XPS spectra of Br 3d, Cs 3d, and Pb 2p (Fig. 2d- resolved PL decay profiles of the CPB@SiO2 samples (Fig. 3e) were
h) are consistent with previous reports on CsPbBr3 [23,32]. The two measured to help better understand the charge carrier dynamics. All PL
peaks of Br 3d3/2 and Br 3d5/2 located at 68.6 and 67.6 eV correspond to decay curves can be fitted with the following double-exponential func­
the inner and surface Br- ions, respectively. The two peaks of Cs 3d3/2 tion [34–36]:

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C.-Y. Zhong et al. Chemical Engineering Journal 431 (2022) 134110

Fig. 2. (a) EDS mapping images showing the elemental distribution of Si, O, Br, Cs, and Pb; the scale bar is 20 nm. (b) EDS linear scanning along the white line
marked in (a). (c) XRD patterns of CPB@SiO2-100 NCs. (d-h) High-resolution XPS spectra of Si 2p, O 1 s, Br 3d, Cs 3d, and Pb 2p for CPB@SiO2-100 NCs.

( ) ( )
− t − t emission peak is red-shifted by ~ 10 nm compared with the one-photon
I(t) = A1 exp + A2 exp (1)
τ1 τ2 PL band shown in Fig. 3b, which can be attributed to photon self-
absorption caused by deeper penetration of NIR photons [41,42]. The
where the short and long decay lifetimes (τ1 and τ2) are attributed to excitation intensity dependence of the PL intensity can be fit with a slope
different decay processes and A1 and A2 embody the homologous of 1.38, 1.67, 1.58, and 1.46 for CPB@SiO2-75, CPB@SiO2-100,
weighted contributions, respectively (Table S2). The average lifetime CPB@SiO2-125, and CPB@SiO2-150, respectively (Fig. 4e). This non-
(τavg) can be obtained by the following equation [37,38]: linear dependence indicates a two-photon absorption mechanism
∑ 2 [2,17,43]. To investigate the carrier dynamics under two-photon exci­
Aτ
τavg = ∑ i i (2) tation, the PL decay profile of CPB@SiO2-100 NCs under 800 nm laser
Ai τ i
was shown in Fig. 4f. The average PL decay lifetime (τavg) is 46.2 ns,
and the calculated τavg values are 33.4, 43.2, 43.8, and 43.6 ns for close to the value under one-photon excitation (43.2 ns). This suggests
CPB@SiO2-75, CPB@SiO2-100, CPB@SiO2-125, and CPB@SiO2-150, that the electronic states involved in both the two-photon and one-
respectively (Fig. 3d). The results indicate that the SiO2 shell increases photon processes are similar. Fig. 5 illustrates the main mechanism of
the observed lifetime, likely by suppressing non-radiative recombina­ light absorption and PL. In the two-photon process, since one NIR
tion of the electron-hole pairs [39,40]. However, more TMOS did not photon (800 nm) with the energy of 1.55 eV is not enough for direct
further increase the observed lifetime. This is consistent with the above inter-band transition (the calculated bandgap of CsPbBr3 is 2.34 eV
PL results, and the optimized silica shell thickness is confirmed to be ~ shown in Fig. S6), two photons are required to excite an electron from
30 nm. the valence band (VB) to the conduction band (CB). After relaxation
The two-photon fluorescence spectra of CPB@SiO2-75, CPB@SiO2- within the bands, the electrons and holes reach their respective band
100, CPB@SiO2-125 and CPB@SiO2-150 samples are shown in Fig. 4a-d. edges, just like a single photo above the bandgap transition, producing
Under the excitation of 800 nm, an emission peak at ~ 526 nm is the green PL peaked at 526 nm. The other electrons and holes recombine
observed, and the emission intensity increases with the laser power. The non-radiatively.

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C.-Y. Zhong et al. Chemical Engineering Journal 431 (2022) 134110

Fig. 3. (a) Absorption spectra of CPB@SiO2 NCs. (b) Photoluminescence spectra of CPB@SiO2 NCs under 365 nm excitation. (c) Photographs of CPB@SiO2 NCs
under sunlight(left) and UV (right), from top to bottom are CPB@SiO2-150, CPB@SiO2-125, CPB@SiO2-100, and CPB@SiO2-75 NCs. (d) Emission peak position and
FWHM of CPB@SiO2 NCs. (e) Time-resolved PL decay profile of CPB@SiO2 NCs under 365 nm. (f) Variation of photoluminescence intensity and average lifetime
along with the amount of TMOS.

Fig. 4. Two-photon fluorescence spectrum of (a) CPB@SiO2-75, (b) CPB@SiO2-100, (c) CPB@SiO2-125, and (d) CPB@SiO2-150 NCs under 800 nm excitation for
several excitation laser power levels. (e) Power dependence of two-photon excited fluorescence intensity of CPB@SiO2 NCs under 800 nm excitation. (f) Time-
resolved PL decay profile of CPB@SiO2-100 NCs under 800 nm excitation.

To verify the protective effect of SiO2, the representative CPB@SiO2- photostability of CPB NCs and CPB@SiO2-100 NCs dispersed in toluene
100 NCs were taken to evaluate the photostability under different con­ solution are compared under the 365 nm UV light irradiation. As shown
ditions. As shown in Fig. S7, the fluorescence intensity of CPB@SiO2-100 in Fig. S8, under long-term irradiation, the fluorescence intensity of CPB
NCs decreases slightly after being stored in air for 6 and 12 months, and NCs decreases significantly, while that of CPB@SiO2-100 NCs is rela­
the fluorescence peak position and the FWHM have no change. It clearly tively stable. In addition, CPB@SiO2-100 NCs can maintain 80% of the
shows that CPB@SiO2-100 NCs have excellent air stability. The original fluorescence intensity after irradiation under 800 nm

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(Fig. 6c), while the experimental group emitted bright green fluores­
cence due to two-photon absorption (Fig. 6d). This indicates that the
cellular luminescence signals originate from CPB@SiO2-100 NCs that
entered into the cells through endocytosis. The cumulative fluorescence
intensity of the experimental group was five times higher than that of
the control group (Fig. 6b). This demonstrates that using CPB@SiO2-100
NCs as the two-photon imaging nanoprobes can effectively avoid the
interference of autofluorescence of biological cells and ensure good
imaging quality.

4. Conclusions
Fig. 5. Schematic diagram of the two-photon absorption and emission process
in CPB@SiO2 NCs. In summary, we have synthesized monodisperse CPB@SiO2 core–­
shell NCs by a modified supersaturated recrystallization method. The
femtosecond laser for 2 h (Fig. S9). The above results indicate solidly optimal thickness of silica shell at ~ 30 nm can alleviate aggregation and
that the CPB@SiO2-100 NCs have good photostability and can be a reduce non-radiative recombination of charge carriers. Under the exci­
candidate material for two-photon photoluminescence bio-probes. tation of a NIR femtosecond laser of 800 nm, the core–shell NCs show
To evaluate the biocompatibility of the CPB@SiO2 NCs, a series of strong two-photon absorption and emit green light peaked at 526 nm,
cytotoxicity experiments was conducted by the standard 3-(4-pyrroli­ with a slope of 1.38 ~ 1.67 for the pumping power dependence of the
dine-5-dimethylthiazolyl-2-yl)-2-pyrrolidine-5-diphenyltetrabrozole two-photon PL intensity. The average PL decay lifetime under two-
(MTT) assay [44,45]. When using CPB@SiO2-100 NCs with a concen­ photon excitation is close to the value under one-photon excitation,
tration of 16 μg/mL for cultivation, the survival rate of HepG2 cells was indicating the same or very similar states involved under both types of
as high as 96.21% (Fig. 6a). Increasing the concentration of CPB@SiO2- excitation. Besides, the survival rate of HepG2 cells maintains 96.21%
100 NCs to 48 μg/mL, the survival rate of the cultured cells still main­ after culturing with the typical CPB@SiO2-100 NCs for 24 h, demon­
tains above 88.83%, which indicates that the NCs have good biocom­ strating excellent biocompatibility. The cumulative fluorescence in­
patibility. To investigate the application potential of CPB@SiO2 NCs in tensity of the two-photon fluorescence images of the experimental group
two-photon fluorescence biological imaging, HepG2 cells were incu­ with CPB@SiO2 NCs is five times higher than the control group. This
bated with CPB@SiO2-100 NCs with a concentration of 16 µg/mL for 24 work demonstrates a reliable method for the preparation of controllable
h. After washing with phosphate buffer saline (PBS) three times, HepG2 perovskite-based core–shell nanostructures with potential applications
cells were scanned by 800 nm femtosecond laser under a laser scanning in two-photon bioimaging.
confocal microscope. Fig. 6c-d show the images of the control group and
the experimental group, which were incubated with or without
CPB@SiO2-100 NCs, respectively. Under the 800 nm laser scanning, the Declaration of Competing Interest
HepG2 cells in the control group showed very weak fluorescence
The authors declare that they have no known competing financial

Fig. 6. (a) Viability of HepG2 cells cultured with CPB@SiO2-100 NCs with varying concentrations for 24 h. (b) Fluorescence brightness of HepG2 cells in the
experimental and control group (incubated with or without CPBr@SiO2-100 NCs). Fluorescence image of HepG2 cells (c) control group and (d) experimental group
under 800 nm illumination.

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interests or personal relationships that could have appeared to influence [19] J. Chen, K. Zidek, P. Chabera, D. Liu, P. Cheng, L. Nuuttila, M.J. Al-Marri,
H. Lehtivuori, M.E. Messing, K. Han, K. Zheng, T. Pullerits, Size- and wavelength-
the work reported in this paper.
dependent two-photon absorption cross-section of CsPbBr3 perovskite quantum
dots, J. Phys. Chem. Lett. 8 (2017) 2316–2321.
Acknowledgments [20] M. Xing, B. Chen, J. Feng, W. Xu, Y. Bai, Y. Zhou, C. Dong, H. Zhong, J. Zhang,
Y. Yin, Confined growth of quantum dots in silica spheres by ion exchange of
“trapped NH4+” for white-light emission, Chem 5 (2019) 2195–2214.
Y. Chen thanks the support of the Natural Science Foundation of [21] N. Ding, D. Zhou, X. Sun, W. Xu, H. Xu, G. Pan, D. Li, S. Zhang, B. Dong, H. Song,
Guangdong Province (Grant No. 2021A1515010152), the Science and Highly stable and water-soluble monodisperse CsPbX3/SiO2 nanocomposites for
white-LED and cells imaging, Nanotechnology 29 (2018), 345703.
Technology Research Project of Guangzhou (No. 201804010047,
[22] Z. Hu, Z. Liu, Y. Bian, S. Li, X. Tang, J. Du, Z. Zang, M. Zhou, W. Hu, Y. Tian,
202002010007), and the Science Research Project of Guangzhou Uni­ Y. Leng, Enhanced two-photon-pumped emission from in situ synthesized
versity (Grant. No. YG2020014). Z. Chen thanks the support of the nonblinking CsPbBr3/SiO2 nanocrystals with excellent stability, Adv. Opt. Mater. 6
(2018) 1700997.
Natural Science Foundation of Guangdong Province (Grant No.
[23] W. Song, Y. Wang, B. Wang, Y. Yao, W. Wang, J. Wu, Q. Shen, W. Luo, Z. Zou,
2020A1515010411). Super stable CsPbBr3@SiO2 tumor imaging reagent by stress-response
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