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Author’s Accepted Manuscript
luminescence thermal sensors operating in the
optical windows of biological tissues
I.E. Kolesnikov, A.A. Kalinichev, M.A.
Kurochkin, D.V. Mamonova, E.Yu. Kolesnikov,
A.V. Kurochkin, E. Lähderanta, M.D. Mikhailov
To appear in: Journal of Luminescence
Received date: 2 May 2018
Revised date: 12 August 2018
Accepted date: 15 August 2018
Cite this article as: I.E. Kolesnikov, A.A. Kalinichev, M.A. Kurochkin, D.V.
Mamonova, E.Yu. Kolesnikov, A.V. Kurochkin, E. Lähderanta and M.D.
Mikhailov, Y 2O3:Nd3+ nanocrystals as ratiometric luminescence thermal sensors
operating in the optical windows of biological tissues, Journal of Luminescence,
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Y2O3:Nd3+ nanocrystals as ratiometric luminescence thermal sensors operating in the
optical windows of biological tissues
I.E. Kolesnikov*,a,b, A.A. Kalinicheva, M.A. Kurochkina, D.V. Mamonovac, E.Yu. Kolesnikovd,
A.V. Kurochkina, E. Lähderantab, M.D. Mikhailovc
St. Petersburg State University, 7/9 Universitetskaya nab., 199034, St. Petersburg, Russia
Lappeenranta University of Technology LUT, Skinnarilankatu 34, 53850, Lappeenranta,
Scientific and Technological Institute of Optical Material Science, VNTs S. I. Vavilov State
Optical Institute, Babushkina 36-1, 192171, St. Petersburg, Russia
Volga State University of Technology, Lenin sqr. 3, 424000, Yoshkar-Ola, Russia
Contact information
e-mail address: (I. Kolesnikov)
Here, we report Nd3+-doped Y2O3 nanoparticles suitable for luminescence thermal sensing in the
first and second biological windows. The nanoparticles were synthesized via the combined
Pechini-foaming method. A ratiometric approach, based on the relative changes in the intensities
of different emission bands corresponding to the Stark sublevels or excited levels, was applied to
determine local temperature. The evaluated thermal sensitivities differed 5-fold times depending
on the choice of transitions for the luminescence intensity ratio calculation. The temperature
uncertainty was determined to be below 1 oC, which allows to perform sub-degree thermal
sensing. The results of ex vivo experiment indicate that Nd3+-doped Y2O3 nanoparticles are
promising candidates for real biological applications.
Keywords: Nd3+, Y2O3, Luminescence, Thermometry, Biological windows
An accurate thermal measurement is highly demanded in many areas from technology to
biomedical field [1–3]. The conventional temperature sensors, such as thermocouples and
thermistors, are unsuitable for remote temperature measurement at the micro and nanoscale [4,5].
Much attention is paid to the development of new classes of non-invasive, non-contact
nanothermometers exhibiting superior spatial and temperature resolution coupled with high
thermal sensitivity [6]. One of the most promising classes of such thermal sensors is
luminescence nanothermometers. Among many different luminescent materials used as noncontact luminescent temperature sensors, inorganic hosts doped with lanthanide ions have
attracted significant interest due to their unique advantages compared to those of the other
luminescent thermometers such as quantum dots, polymers, and organic phosphors. In particular,
these phosphors exhibit high chemical, mechanical, and thermal stability, perfect photostability,
no photo blinking effects, as well as demonstrate wide usable temperature sensitivity range [7,8].
Real-time temperature sensing during hyperthermia treatment is of crucial importance to reduce
the overheating side effects. To perform in vivo thermal sensing, excitation and emission bands
of luminescent thermometer have to lie in spectral region where transparency of living tissues is
high due to low optical absorption [9,10]. There are three such regions, so-called biological
optical transparency windows (BW): 750-950 nm (I-BW), 1000-1350 nm (II-BW), 15001
1800 nm (III-BW), which contributes to the deep tissue imaging prospects in a different way
[11,12]. In terms of imaging, the I-BW can suffer from the interference of tissue
autofluorescence [13,14] nevertheless the exceptional tissue penetration properties of the I-BW
are commonly exploited for effective NIR laser excitation [15]. The II-BW and III-BW are
significantly better suited for NIR imaging at greater tissue depths [16], due to extremely
reduced NIR light scattering [17,18]. Thus, ideal luminescent nanothermometer should be
excited in the I-BW and emit light in the II-BW or III-BW [19].
Nd3+ ions perfectly fit aforementioned requirements having shielded ladder-like intra-4f levels.
Nd3+-doped nanoparticles (NPs) can be effectively excited with NIR light of around 800 nm
wavelength, which avoids undesired heating of the surrounding [20,21] and subsequent faulty
temperature readout. Additionally, Nd3+-doped NPs demonstrate quite intensive emission lines
situated in all three BW, which broaden possible application in deep tissue in vivo imaging and
nanothermometry [15,22,23].
Usually manuscripts devoted to the ratiometric Nd3+-doped thermometers are focused on the
single temperature dependent parameter [24–26]. In this work, we report Y2O3:Nd3+ NPs to
demonstrate luminescent temperature sensing in the first and second biological windows upon
NIR excitation using various luminescence intensity ratios. The important parameters for
nanothermometry such as thermal sensitivity and temperature uncertainty have been calculated
and compared with other Nd3+-doped sensors. The potential in vivo application of Y2O3:Nd3+
NPs has been explored by carrying out ex vivo experiments.
Nd3+-doped Y2O3 nanoparticles was synthesized by the combined Pechini-foaming
method. Unlike previously developed modified Pechini technique [27,28], this method
also reduces agglomeration of derived particles but does not require second calcination
procedure in molten salt. The main part of modification in the combined Pechini-foaming
method is implementation of foaming agent into polymer gel matrix in order to prevent
strong agglomeration of synthesized samples. The foaming agent is mixture of aluminium
nitrate and potassium chloride.
Synthesis of Eu3+ and Nd3+-doped oxides using the combined Pechini-foaming method
were described in details in our previous papers [29–31]. Therefore, only short
description of Y2O3:Nd3+ 1 at.% nanocrystalline phosphors synthesis is presented.
Briefly, aluminium nitrate was added to the yttrium and neodymium nitrates in molar
ratio Al(NO3)3:Y(NO3)3=5:3. Then saturated solution of citric acid in distilled water
(volume ratio 1:1) and potassium chloride powder were successively added to the
abovementioned solution. The resultant mixture was heated up to 150–200 oC leading to
the chelate complexes formation. The acid excess reacts with the salt, which results in the
potassium dihydrogen citrate formation; oxidized, it forms potassium carbonate. Adding
the ethylene glycol in the solution leads to the etherification reaction with formation of
the polymer gel as a result. The polymer gel contains aluminium, yttrium and europium
ions which are uniformly distributed in polymer network and potassium carbonate which
are situated in polymer cells. In order to remove organic components, the gel is calcinated
at 1000 oC for 2 hours. Uniformly distributed aluminium oxide reacts with potassium
carbonate, which results in formation of KAlO2 accompanied by intense gas release. Y2O3
nanoparticles with homogeneously distributed Nd3+ ions are obtained, whereas reaction
byproducts are removed by washing in distilled water. The synthesized nanoparticles
were collected by centrifugation at 2800 rpm for 5 min and then dried naturally.
X-ray diffraction patterns were registered with the powder diffractometer UltimaIV (Rigaku) in
Bregg-Bretano geometry with CuKα1 radiation (λ = 1.54059 Å) in the 2θ range from 10o to 80o.
Electron micrograph images were obtained with SUPRA 40VP WDS scanning electron
microscope (resolution 4 nm). Particle size was measured with laser diffraction particle size
analyzer Mastersizer 3000 using static light scattering technique. Fluorescence characterization
was performed with fluorescence spectrometer Fluorolog-3 and T64000 Raman Spectrometer.
Nd3+-doped Y2O3 NPs was optically excited with a 808 nm single mode laser Coherent MBR110 (5 mW) and 532 nm solid state laser Thorus 532 (1 mW). The laser beam was focused into
the sample by using a 4x long working distance microscope objective (NA 0.1). The
fluorescence was collected by using the same microscope objective and was spectrally analyzed
by single spectrometer and a liquid nitrogen cooled Symphony II CCD detector. Thermal
sensitivity and uncertainty experiments were carried out using heating stage controlled with
ThorLabs TC200 with a resolution of 0.1 °C. Sub-tissue temperature sensing was performed with
chicken breast as a phantom of human tissue. 50 µl of aqueous colloidal solution of Nd3+-doped
Y2O3 NPs (5 mg mL−1) was injected in fresh chicken breast at the different depths. The colloidal
solution was prepared by dispersing powder particles in deionized water with further sonification
for 5 min.
Results and discussion
Fig. 1a presents XRD pattern of Y2O3:Nd3+ 1 at.% nanocrystalline phosphors which
demonstrates presence of Y2O3 as a single phase without any impurities. All diffraction patterns
coincide with cubic phase of Y2O3 (JCPDS 41-1105). Fig. 1b shows scanning electron
microphotograph of the synthesized Y2O3:Nd3+ 1 at.% powder. As seen from the micrograph, the
powder consists of weakly agglomerated nanoparticles with average size about 40–50 nm. The
particle size distribution of Y2O3:Nd3+ 1 at.% powder determined using static light scattering
(SLS) technique demonstrates the mean size at about 54 nm (Fig. 1c), which indicates a good
correlation between SLS and SEM data.
Figure 1. (a) XRD pattern of Y2O3:Nd3+ 1 at.% and standard card of Y2O3 (JSPDS 41-1105);
(b) SEM image of Y2O3:Nd3+ 1 at.% nanoparticles with particle size distribution as insert; (c)
particle size distribution in aqueous solution of Y2O3:Nd3+ 1 at.% determined using static light
Excitation spectrum of Y2O3:Nd3+ 1 at.% NPs is shown in Fig. 2a. Spectrum measured in the
spectral range 350–850 nm consists of intra-configurational f-f transitions from ground level 4I9/2
to excited levels in Nd3+ ion. During scanning excitation wavelength luminescence signal was
monitored at the prominent transition 4F3/2 – 4I11/2 (λem=1079.4 nm). Excitation bands were
observed at 350–382, 420–445, 455–495, 500–557, 560–642, 665–705, 710–780, 780–850 nm.
They were ascribed to the transitions of 4I9/2 – 4D3/2+ 4D1/2, 4I9/2 – 2D5/2, 4I9/2 – 4G9/2+4G11/2+2K15/2,
I9/2 – 4G7/2+4G9/2+2K13/2, 4I9/2 – 4G5/2+4G7/2+2H11/2, 4I9/2 – 4F9/2, 4I9/2 – 4F7/2+4S3/2, 4I9/2 – 4F5/2+4H9/2
respectively [32]. In this paper, luminescence of Y2O3:Nd3+ NPs was excited by 808 and 532 nm,
which is schematically shown in Fig. 2a.
Emission spectrum of nanocrystalline Y2O3:Nd3+ powder upon 808 nm is presented in Fig. 2b. It
should be noted that all emission spectra presented in this work were not corrected for the
detector response. Measured spectrum consisted of characteristic narrow lines attributed to the
transitions from excited 4F3/2 to the lower 4IJ levels: 4F3/2 – 4I9/2 (870–985 nm) and 4F3/2 – 4I11/2
(1042–1160 nm). Each transition consists of several lines corresponding to the Stark splitting of
excited and ground levels due to crystal field. Emission spectrum is dominated by 4F3/2(1) –
I11/2(2) transition centered at 1079.4 nm. However, it should be noted that intensive
luminescence bands were observed in both I-BW and II-BW.
Figure 2. (a) Excitation and (b) emission spectra of Y2O3:Nd3+ 1 at.% nanoparticles.
Nowadays, ratiometric Nd3+ based NPs provide two different methods for thermal sensing. The
first one is based on the analysis of emission bands corresponding to the transitions between
Stark sublevels of 4F3/2(Ri) and 4I9/2(Xj) or 4I11/2(Yk) energy levels.
Detailed emission spectra of Y2O3:Nd3+ 1 at.% NPs at different temperatures (26.5 and 58.5 oC)
measured in the spectral range of 870–922 nm (I-BW) are presented in Fig. 3a. The
luminescence intensity ratio between 4F3/2(2) – 4I9/2(3) and 4F3/2(1) – 4I9/2(3) (hereafter denoted as
1 ) and between 4F3/2(2) – 4I9/2(1) and 4F3/2(1) – 4I9/2(1) (hereafter denoted as 2 ) were chosen for
nanothermometry (Fig. 3b), because they should have temperature dependence in biological
range due to the value of energy gap between Stark sublevels. As can be seen, the emission lines
used for thermal sensing are well resolved, so deconvolution analysis is not required. According
to previous study, luminescence intensity ratio (LIR) was calculated using integral intensities
(integration width was about FWHM) instead of peak intensities, resulting in the better thermal
sensitivity and temperature uncertainties [33]. Both obtained LIRs demonstrated monotonous
pseudo-linear trend within studied temperature range (Fig. 3c, 3d).
Figure 3. (a) Emission spectra of Y2O3:Nd3+ 1 at.% nanoparticles obtained at two different
temperatures (λex=808 nm); (b) energy levels scheme of transition 4F3/2(Ri) – 4I9/2(Xj) for Nd3+
ion in the Y2O3 host; (c), (d) luminescence intensity ratios 1 and 2 as a function of
temperature. Red lines correspond to the best fitting.
Thermal sensing using Y2O3:Nd3+ 1 at.% NPs was performed also in the II-BW. Detailed
emission spectra at different temperatures (26.5 and 58.5 oC) measured in the spectral range of
1040–1090 nm are presented in Fig. 4a. Nanothermometry was based on the luminescence
intensity ratio between 4F3/2(2) – 4I11/2(1) and 4F3/2(1) – 4I11/2(1) (hereafter denoted as 1 ) and
between 4F3/2(2) – 4I9/2(2) and 4F3/2(1) – 4I11/2(2) (hereafter denoted as 2 ) (Fig. 4b). Fig. 4c and
4d showed aforementioned LIRs as a function of temperature from which a pseudo-linear
behavior was observed.
Figure 4. (a) Emission spectra of Y2O3:Nd3+ 1 at.% nanoparticles obtained at different
temperatures (λex=808 nm); (b) energy levels scheme of transition 4F3/2(Ri) – 4I11/2(Yk) for
Nd3+ion in the Y2O3 host; (c), (d) luminescence intensity ratios 1 and 2 as a function of
temperature. Red lines correspond to the best fitting.
It should be noted that the adjusted R2 were found to be 0.99 for all luminescence intensity ratios
in both I-BW and II-BW. So, the obtained thermal calibrations could provide information about
local temperature around Y2O3:Nd3+ NPs.
Besides thermal sensing using 4F3/2(Ri) – 4I9/2(Xj) or 4F3/2(Ri) – 4I11/2(Yk) Stark components, an
alternative method relies on LIR between 4F5/2(4) – 4I9/2(3) (820 nm) and 4F3/2(1) – 4I9/2(1)

(892 nm) transitions (532
) (Fig. 5a). Due to the higher energy difference between excited levels
used for nanothermometry, significant enhancement of the relative thermal sensitivity should be
observed. Moreover, this LIR can be potentially used for thermal sensing at high temperature. In
this case the luminescence of Y2O3:Nd3+ 1 at.% NPs was excited with 532 nm radiation (Fig.
5b). Emission spectra consist of narrow lines attributed to the following transitions: 4F5/2(4) –
I9/2(3) (820.4 nm), 4F5/2(2) – 4I9/2(3) (829 nm),4F5/2(3) – 4I9/2(4) (838.4 nm), 4F3/2(2) – 4I9/2(1)
(876.7 nm), 4F3/2(2) – 4I9/2(2) (878.8 nm), 4F3/2(1) – 4I9/2(1) (892.3 nm), 4F3/2(1) – 4I9/2(2)
(894.6 nm), 4F3/2(2) – 4I9/2(3) (897.4 nm), 4F3/2(1) – 4I9/2(3) (913.8 nm), 4F3/2(2) – 4I9/2(5)
(929.4 nm), 4F3/2(1) – 4I9/2(5) (946.4 nm). It should be noted that luminescence intensity of
emission lines assigned to the transitions from 4F5/2 level is much lower than from 4F3/2 one and
the corresponding part of spectrum is multiplied by 20. As it can be seen, the intensity of 4F5/2 –
I9/2 transitions increased with temperature increase, whereas the intensity of 4F3/2 – 4I9/2
transitions reduced. Dependence of LIR on the temperature within biological range is shown in
Fig. 5c. The thermal calibration was fitted with linear function similar to other luminescence
intensity ratios.
Figure 5. (a) Energy levels scheme of transition 4F5/2 – 4I9/2 and 4F3/2 – 4I9/2 for Nd3+ion in the
Y2O3 host; (b) emission spectra of Y2O3:Nd3+ 1 at.% nanoparticles obtained at different

temperatures (λex=532 nm); (c) luminescence intensity ratios 532
as a function of temperature.
Red line corresponds to the best fitting.
An absolute thermal sensitivity expresses the suitability of the luminescent thermometer for
temperature sensing. However, in order to compare the thermal performances between different
nanothermometers irrespective of their nature, it is very useful to evaluate the relative thermal
sensitivity Srel, which can be expresses as follows:
∙ 100%
where R is obtained LIR. The relative thermal sensitivities of Y2O3:Nd3+ 1 at.% NPs calculated
for Stark sublevels and 4F5/2, 4F3/2 levels LIRs are listed in Table 1. Table 1 includes also the
relative thermal sensitivities of different Nd3+-based NPs as well as excitation wavelength, LIR
transition and thermal sensing range. Careful examination of Table leads to conclusion that
among NIR-to-NIR nanothermometers Y2O3:Nd3+ nanophosphor demonstrates the best relative
thermal sensitivity. It should be noted that thermal sensitivity in the I-BW showed lower value
than in the II-BW. Thermal sensing based on LIR between 4F5/2 – 4I9/2 and 4F3/2 – 4I9/2 transitions
demonstrates almost 5-fold enhancement of sensitivity compared with Stark sublevels. The
obtained value of 1.59 % oC-1 is close to the maximum relative sensitivities 1.75±0.04 % oC-1 and
1.8 % oC-1 attained for Gd2O3:Nd3+ [34] and YAlO3:Nd3+ NPs [35], respectively. However, it
should be emphasized that emission of Gd2O3:Nd3+ NPs was measured with R928
photomultiplier, whereas in this work a simple charge coupled device (CCD) InGaAs-based
detector was used.
Table 1. Comparison of Nd3+-doped nanoparticles as ratiometric thermometers.
LIR transitions
Temperature Srel, (% oC-1)
range (oC)
808 4F3/2 – 4I9/2 (Stark sublevels, 1 )

F3/2 – I9/2 (Stark sublevels, 2 )
F3/2 – I11/2 (Stark sublevels, 1 )

F3/2 – I11/2 (Stark sublevels, 2 )

F5/2, F3/2 – I9/2 (532 )
F3/2 – I9/2 (Stark sublevels)
F3/2 – I9/2 (Stark sublevels)
F3/2 – I9/2 (Stark sublevels)
F3/2 – I9/2 (Stark sublevels)
F3/2 – I9/2 (Stark sublevels)
F3/2 – I9/2 (Stark sublevels)
F3/2 – I9/2 (Stark sublevels)
F3/2 – I11/2 (Stark sublevels)
KGd(WO4)2 808
F3/2 – I9/2 (Stark sublevels)
F3/2 – I11/2 (Stark sublevels)
F3/2 – I11/2 / F3/2 – I9/2
(Stark sublevels)
580 4F5/2, 4F3/2 – 4I9/2
F5/2, F3/2 – I9/2
F5/2, F3/2 – I9/2
If the relative sensitivity is used to compare the performance of different thermometers, the
temperature uncertainty, ΔT, defines the accuracy of temperature evaluation using
nanothermometer and specific experimental setup. As it was shown in our previous paper [33],
the temperature uncertainty can be calculated with various methods. Here, it was obtained from
acquisition of several consecutive emission spectra at a fixed temperature. 50 emission spectra of
Y2O3:Nd3+ 1 at.% NPs were measured at heating stage with controlled temperature (30 oC). The
statistical distributions of temperature determined from luminescence thermometry were fitted
by Gauss function and FWHM was used as an estimation of the temperature uncertainty. The

temperature uncertainties were calculated for LIRs with the highest thermal sensitivities (532
1 and 2 ), and the obtained results are presented in Fig. 6. The temperature uncertainties are
below 1 oC, which makes Nd3+-doped Y2O3 NPs perspective candidates for thermal sensing with
sub-degree resolution.
Figure 6. Statistical distributions calculated from various luminescence intensity ratios
(Theater = 30 oC).
It should be noted that the temperature uncertainty can be decreased by improving signal-tonoise ratio by increasing acquisition time and/or by averaging consecutive measurements of the
emission spectrum. Usually thermal sensing in real applications requires fast measurement, so
one must have a compromise between decreasing the acquisition time and lowering the
temperature uncertainty. If acquisition time in certain type of measurement should not be
minimized, the minimum achievable temperature uncertainty is defined by the uncertainty of the
experimental setup, in the order of ∼0.0001% for the case of a laboratory-grade
spectrofluorimeter [34].
To probe the potential use of Nd3+-doped Y2O3 NPs for temperature changes investigation in
biological systems, simple ex vivo experiment has been carried out. We injected 50 µl of aqueous
colloidal solution of Y2O3:Nd3+ 1 at.% NPs (5 mg mL−1) in fresh chicken breast at the different
depths. Chicken breast was used to mimic optical properties of human tissue. Experimental setup
is schematically presented in Fig. 7a. Briefly, low intensity 808 nm laser beam excited emission
of NPs through chicken breast. Laser intensity was kept minimal (5 mW) to avoid induced
heating effects. Optical excitation and subsequent luminescence collection were both performed
by using a single long working distance 4x objective. Spectral analysis and LIR calculation of
the sub-tissue luminescence signal was used to determine the injection’s temperature. Taking
into account extinction coefficient of tissue and thermal sensitivity of studied LIR, sub-tissue
temperature sensing was performed in the II-BW using 1 intensity ratio. The temperatures
obtained for different tissue depths and their errors are presented in Fig. 7b. The average
temperature was found to be 26.3 oC, which is very close to the value measured with
thermocouple (26.5 oC). It should be noted that increasing of tissue depth significantly increases
thermal sensing error. The provided experiments demonstrated the applicability of Nd3+-doped
Y2O3 NPs for temperature determination purposes in ex vivo biomedical studies with a good
thermal resolution.
The carried out study opens an avenue toward controllable photothermal treatment, which is
crucial to reduce collateral damage. Bio-functionalized Nd3+-doped Y2O3 NPs can be utilized to
search temperature singularities in vivo for diagnostic purposes. Noteworthy, along with submicrometric spatial and sub-degree thermal resolution use of ratiometric luminescence
thermometers and CCD camera as a detector provide real-time sensing, so local temperature can
be defined with sub-second temporal resolution. All these features are of great importance in real
biological and medical applications.
Figure 7. (a) Scheme of the experimental setup for ex vivo temperature sensing; (b) calculated
temperature as a function of tissue depth.
In this work, Nd3+-doped Y2O3 nanoparticles have been synthesized by the combined Pechinifoaming method. Single phase Y2O3:Nd3+ 1 at.% phosphor consists of nanoparticles with average
size about 50 nm, which was determined with SEM and SLS measurements. Two different
approaches were applied to provide ratiometric thermal sensing: 1) LIR was based on the
analysis of emission bands corresponding to the transitions between Stark sublevels of 4F3/2(Ri)
and 4I9/2(Xj) or 4I11/2(Yk); 2) LIR was calculated between transitions from different excited
energy levels (4F5/2 – 4I9/2 and 4F3/2 – 4I9/2). The main advantage of the first approach is to have
both excitation and emission lines in the biological windows, which plays crucial role in
biological and medicine applications, whereas the second approach enhances thermal sensitivity
5-fold but requires using 532 nm excitation. The temperature uncertainty obtained from
acquisition of several consecutive emission spectra at fixed temperature was determined to be
below 1 oC. This makes Nd3+-doped Y2O3 NPs perspective candidates for thermal sensing with
sub-degree resolution. Ex vivo measurements conducted with chicken breast proved possibility
of luminescence temperature sensing in the II-BW through 3 mm tissue.
This research has been supported by the Russian Science Foundation (№ 17-72-10055).
Experimental investigations were carried out in “Center for Optical and Laser materials
research“, “Research Centre for X-ray Diffraction Studies“, “Interdisciplinary Resource Center
for Nanotechnology”, and “Innovative Technologies of Composite Nanomaterials“ (St.
Petersburg State University).
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