5604 Langmuir 2006, 22, 5604-5610

A General Method for the Controlled Embedding of Nanoparticles in

Silica Colloids
Christina Graf,* Sofia Dembski, Andreas Hofmann, and Eckart Rühl*,†
Institut für Physikalische Chemie, UniVersität Würzburg, Am Hubland, D-97074 Würzburg, Germany

ReceiVed January 14, 2006. In Final Form: April 12, 2006

A novel method for the controlled embedding of multiple nanoparticles of various materials, such as gold nano-
particles, quantum dots, and magnetic nanoparticles, in silica colloids is presented. After adsorption of the amphiphilic
polymer poly(vinylpyrrolidone) on hydrophobic or hydrophilic stabilized nanoparticles, these are adsorbed on silica
spheres and covered by variable-thickness silica shells. This silica coating protects the embedded nanoparticles against
chemical transformations, which is of crucial importance for the biocompatibility of particles containing toxic elements.
Moreover, it is found that the optical properties of the nanoparticles are retained. Possible applications of multicore
particles are briefly discussed.

Introduction
Nanoparticles have unique, size-dependent properties. There-
fore, they are of increasing interest for both fundamental research
and numerous applications. Small noble metal nanoparticles
exhibit characteristic colors due to the excitation of surface
plasmon modes, which can be tuned by their size, shape, and the
local environment. Semiconductor nanoparticles (quantum dots,
QD) are of specific interest because of their specific optical and
electronic properties.1-3 This is accompanied by a sharp, tunable
emission band, where strong resistance to photobleaching is
observed. As a consequence of these distinct properties, QD
have the potential for numerous applications including biolabeling
or they may be used as tracers in diffusion studies. Superpara-
magnetic nanoparticles have been studied with respect to
biomedical applications,4-6 such as MRI contrast enhancement,7
magnetic immobilization, and drug targeting.8
Many of the promising applications of nanoparticles require
that they are transferred in a suitable medium, for instance, in
aqueous solution allowing for applications in life sciences. High-
quality semiconductor nanoparticles are usually stabilized by
organic ligands that render them hydrophobic. However, they
are often unstable with respect to oxidation and they are cytotoxic.9
Other syntheses yield hydrophilically stabilized particles, e.g.,
citrate-stabilized gold particles that cannot be transferred in apolar
media.10 Therefore, several approaches have been developed
where nanoparticles are coated by protective shells so they can
be transferred in other media.10,11 Usually, such syntheses are
* To whom correspondence should be addressed. E-mail: cgraf@
phys-chemie.uni-wuerzburg.de (C.G.); ruehl@chemie.fu-berlin.de (E.R.).
† Present address: Physikalische und Theoretische Chemie, Institut für
Chemie und Biochemie, Freie Universität Berlin, Takustr. 3, D-14195 Berlin,
Germany.
(1) Empedocles, S. A.; Norris, D. J.; Bawendi, M. G. Phys. ReV. Lett. 1996,
77, 3873-3876.
(2) Norris, D. J.; Bawendi, M. G. Phys. ReV. B 1996, 53, 16338.
(3) Colvin, V. L.; Schlamp, M. C.; Alivisatos, A. P. Nature 1994, 370, 354
(4) Meldrum, F. C.; Heywood, B. R.; Mann, S. Science 1992, 257, 522.
(5) Tanaka, T.; Matsunaga, T. Anal. Chem. 2000, 72, 3518.
(6) Matsunaga, T.; Kawasaki, M.; Yu, X.; Tsujimura, N.; Nakamura, N. l.
Anal. Chem. 1996, 68, 3551.
(7) Harisinghani, M. G.; Barentsz, J.; Hahn, P. F.; Deserno, W. M.; Tabatabaei,
S.; Hulsbergen van de Kaa, C.; de la Rosette, J.; Weissleder, R. New Eng. J. Med.
2003, 348, 2491.
(8) Nakamura, N.; Burgess, J. G.; Yagiuda, K.; Kudo, S.; Sakaguchi, T.;
Matsunaga, T. Anal. Chem. 1993, 65, 2030.
(9) Derfus, A. M.; Chan, W. C. W.; Bhatia, S. N. Nano Lett. 2004, 4, 11
(10) Liz-Marzan, L. M.; Giersig, M.; Mulvaney, P. Langmuir 1996, 12, 4329.
specific to the class of materials of interest, so that these are not
suitable for generally applicable synthesis routes.
The regime of the strongest size effects in optical properties
is confined to 2-40 nm particles. By adsorbing or embedding
such particles in larger matrix spheres, larger particles can be
prepared that still exhibit optical properties of small nanospheres,
but these are considerably easier to handle. This, however, requires
a well-controlled preparation method, especially the prevention
of uncontrolled aggregation and clustering, so that specific
properties of the small nanoparticles are retained. Moreover, it
should be also possible to enhance specific properties in this
way, for instance, by embedding fluorescing particles in a well-
controlled radial distance to, e.g., a metal particle in one larger
colloidal particle and enhancing in this way their fluorescence
quantum yield as it has been already shown for layered
nanoparticle systems.12 This concept may also be used as a better-
defined way for the combination of the properties of two species
in one matrix colloid, such as magnetic and strongly luminescent
properties. Up to now, synthetic routes have been used for this
purpose where the matrix colloid formation and the nanoparticle
inclusion happen simultaneously. Therefore, the interparticle
distances of the embedded nanoparticles can be hardly
controlled.13-15 Moreover, in the embedding process which uses
a defined matrix particle as its core, it appears also to be possible
to control the location of photoluminescing nanoparticles on the
nanometer scale, which allows one to study the local density of
states with respect to photonic applications.
Further, the incorporation of luminescent semiconductor
nanocrystals into photonic crystals has received considerable
attention in the past years as a promising pathway to novel light
sources with controllable spontaneous emission.16 Such crystals
can be prepared from monodisperse latex or silica spheres, either
bare, doped, or covered by a shell. Therefore, the synthesis of
semiconductor nanoparticle-doped colloidal particles that can
(11) Gerion, D.; Pinaud, F.; Williams, S. C.; Parak, W. J.; Zanchet, D.; Weiss,
S.; Alivisatos, A. P. J. Phys. Chem. B 2001, 105, 8861.
(12) Kulakovich, O.; Strekal, N.; Yaroshevich, A.; Maskevich, S.; Gaponenko,
S.; Nabiev, I.; Woggon, U.; Artemyev, M. Nano Lett. 2002, 2, 1449.
(13) Kim, J.; Lee, J. E.; Lee, J.; Yu, J. H.; Kim, B. C.; An, K.; Hwang, Y.;
Shin, C.-H.; Park, J.-G.; Kim, J.; Hyeon, T. J. Am. Chem. Soc. 2006, 128, 688.
(14) Yi, D. K.; Selvan, S. T.; Lee, S. S.; Papaefthymiou, G. C.; Kundaliya,
D.; Ying, J. Y. J. Am. Chem. Soc. 2005, 127, 4990.
(15) Müller-Schulte, D.; Schmitz-Rode, T.; Borm, P. J. Magn. Magn. Mater.
2005, 293, 135.
(16) Rogach, A. L.; Kotov, N. A.; Koktysh, D. S.; Susha, A. S.; Caruso, F.
Colloids Surf., A 2002, 202, 135.

10.1021/la060136w CCC: $33.50 © 2006 American Chemical Society

Published on Web 05/24/2006
Embedding of Nanoparticles in Silica Coloids
serve as building blocks for such crystals is an important issue
in colloidal science.
Particles containing multiple nanoparticles can be prepared
by aggregation processes, such as trapping of nanoparticles during
a sol gel process,17 micelle formation,18 or by controlled adsorption
of the nanoparticles on a preformed supporting core19 and further
coating with a protective layer.20,21 Aggregation processes are
often poorly controllable, and therefore, they yield particles where
neither the polydispersity of the colloidal particles nor the radial
position of the nanoparticles in the matrix particle can be well
controlled. On the other hand, the second method using a
preformed colloidal core is easier to control. However, this often
requires a functionalization step to attach the nanoparticles on
the core colloid or to transfer the nanoparticles into a more polar
medium where the reaction takes place. Both can be achieved
by a ligand-exchange reaction. The success of such a reaction
often depends significantly on the specific surface properties of
the nanoparticle. Consequently, these reactions are often not
well reproducible and they are often limited to certain material
systems. Therefore, the direct adsorption of an appropriate
polymer, such as poly(vinylpyrrolidone) (PVP), on nanoparticles
leading to changes in surface properties and particle stability is
a simple alternative to these exchange reactions.22 The am-
phiphilic, nonionic polymer PVP is widely used in science and
technology and adsorbs readily to a broad range of different
materials including metals (e.g., gold, silver, iron)22 and metal
oxides (kaolinite, titanium dioxide, iron oxide, alumina).23 PVP
stabilizes colloidal particles in water and many nonaqueous
solvents. An exchange of the ligands is in this case not required.
Silica is an attractive material for the embedding of nano-
particles because it can be easily functionalized. In this way, a
wide variety of coating procedures has been developed for silica
colloids that allow one to disperse them in various solvents ranging
from very polar to apolar ones.24 Moreover, several routes for
biofunctionalization of silica colloids are reported.25,26 Silica
colloids are prepared with a very low polydispersity (<1%) by
the Stöber synthesis.27 This preparation method and the further
growth of silica on silica or other vitreophilic seed particles in
an ethanol/ammonia mixture, the so-called Stöber growth, yields
smooth surfaces since the growth takes place on a molecular
scale.28-30 If silica-coated particles are grown further by this
procedure, the polydispersity of the particles decreases with R-1,
where R is the particle radius.30 This permits the growth of crystals
of pure silica and silica core shell particles which are suitable
for photonic applications. Furthermore, is has been shown that
the dense silica network can effectively prevent sensitive species
from oxidation.31
We present a novel method for the controlled embedding of
multiple nanoparticles of different materials in silica colloids in
(17) Mokari, T.; Sertchook, H.; Aharoni, A.; Ebenstein, Y.; Avnir, D.; Banin,
U. Chem. Mater. 2005, 17, 258
(18) Chen, C.; Rosenzweig, Z. Nano Lett. 2002, 2, 1299
(19) Caruso, F. AdV. Mater. 2001, 13, 11
(20) Pastoriza-Santos, I.; Gómez, D.; Pérez-Juste, J.; Liz-Marzan, L. M.;
Mulvaney, P. Phys. Chem. Chem. Phys. 2004, 6, 5056
(21) Gómez, D.; Pastoriza-Santos, I.; Mulvaney, P. Small 2005, 1, 238
(22) Graf, C.; Vossen, D. L. J.; Imhof, A.; van Blaaderen, A. Langmuir 2003,
19, 6693.
(23) Pattanaik, M.; Bhaumik, S. K. Mater. Lett. 2000, 44, 352.
(24) Ketelson, H. A.; Brook, M. A.; Pelton, R. H. Polym. AdV. Technol. 1995,
6, 335.
(25) Liu, S. H.; Han, M. Y. AdV. Funct. Mater. 2005, 15, 961
(26) He, X. X.; Wang, K. M.; Tan, W. H.; Li, J.; Yang, X. H.; Huang, S. S.;
Li, D.; Xiao, D. J. Nanosci. Nanotechnol. 2002, 2, 317.
(27) Stöber, W.; Fink, A.; Bohn, E. J. Colloid Interface Sci. 1968, 26, 62.
(28) Philipse, A. P. Colloid Polym. Sci. 1988, 266, 1174.
(29) Bogush, G. H.; Zukosi, C. F. J. Colloid Interface Sci. 1991, 142, 19.
(30) van Blaaderen, A.; van Geest, J.; Vrij, A. J. Colloid Interface Sci. 1992,
154, 481.
(31) Ohmori, M.; Matijevic, E. J. Colloid Interface Sci. 1993, 160, 288.
Langmuir, Vol. 22, No. 13, 2006 5605
Figure 1. Diagram of the general procedure for embedding
nanoparticles in silica colloids: In the first step, poly(vinylpyrroli-
done) (PVP) is adsorbed on the colloidal particles (1). After transfer
in an alcohol, e.g., ethanol or butanol, the particles are adsorbed on
amino-functionalized silica colloids (2). Finally, after addition of
ammonia, a silica shell is grown on the nanoparticle-decorated colloid
by consecutive additions of tetraethoxysilane (3).
the range from 40 to 200 nm. In a first step, the amphiphilic
polymer PVP is reversibly adsorbed on hydrophobically or
hydrophilically stabilized nanoparticles. These functionalized
particles are subsequently homogeneously adsorbed on the surface
of amino-functionalized silica spheres. The nanoparticle-
decorated colloids are directly covered by a smooth silica shell
of adjustable thickness by a Stöber-like growth process. The
particle size and structure of the multicore particles are
characterized by electron microscopy. The influence of the
polymer coating and the silica environment on the optical and
magnetic properties of the nanoparticles is characterized by
electron and confocal laser scanning microscopy, extinction, and
photoluminescence spectroscopy, NMR, and ICP-MS.
Experimental Section
Materials. 3-Aminopropyltrimethoxysilane (APS, g97%), tetra-
methylammonium hydroxide pentahydrate (97%), CdO (99.99+%),
Se powder (99.99%), hexamethyldisilathiane (S(TMS)2), hexa-
decylamine (HDA, 98%), trioctylphosphine oxide (TOPO, 99%),
methanol (anhydrous, 99.8%), chloroform (anhydrous, g99%), and
stearic acid (95%) were purchased from Aldrich. Dimethylzinc
(ZnMe2, 1 M solution in heptane), tetraethoxysilane (TES, g98.0%),
citric acid (g99.5%), and trioctylphosphine (TOP, 90%) were
obtained from Fluka. Igepal CO-520, PVP with an average molar
mass of 10 kg/mol (PVP-10), was purchased from Sigma. Ammonia
(29.3 wt % NH3 in water) was obtained from Merck, ethanol (p.a.)
from Riedel-de Haën, and cyclohexane (99.5%) from Grüssing. All
chemicals were used as received without any further purification.
Syntheses. The general procedure for the controlled embedding
of multiple nanoparticles in silica colloids is a multiple-step reaction.
An outline of the synthesis is schematically shown in Figure 1.
TOPO-functionalized CdSe/ZnS nanoparticles (QD) of 1.6 nm
radius in chloroform were obtained from syntheses based on modified
versions of literature methods.32,33 These are purified by several
washing steps in methanol/chloroform (final concentration 5 g/L in
chloroform). Aqueous dispersions of gold colloids of 9 nm radius
(c ) 0.098 g/L) were synthesized according to the standard sodium
citrate reduction method.34,35 Aqueous dispersions of maghemite
(32) Qu, L. H.; Peng, X. G. J. Am. Chem. Soc. 2002, 124, 2049.
(33) Hines, M. A.; Guyot-Sionnest, P. J. Phys. Chem. 1996, 100, 468.
(34) Enüstün, B. V.; Turkevich, J. J. Am. Chem. Soc. 1963, 85, 3317.
(35) Frens, G. Nat. Phys. Sci. 1973, 241, 20.
5606 Langmuir, Vol. 22, No. 13, 2006
nanoparticles of 4.5 nm radius (c ) 21 g/L) were prepared by a
method described by Massart36 and adjusted to a pH of 7 by the
addition of 0.01 M citric acid and a concentrated solution of
tetramethylammonium hydroxide pentahydrate in water, as described
by van Ewijk et al.37 The gold and maghemite nanoparticle dispersions
were not further purified. Silica particles (final radius 35, 106, and
192 nm) were prepared starting from seed particles obtained from
a microemulsion synthesis38 or Stöber synthesis.27 These are
subsequently grown larger by seeded growth to the final radii.28-30
In the next step, they were functionalized with amino groups as
described in refs 39 and 40. (see Supporting Information).
The amount of PVP was calculated to yield colloids with about
60 PVP molecules per square nanometer of surface. PVP solutions
were prepared by dissolving 27.1 g PVP/L in butanol (in the case
of the QD) or water (in the case of gold and maghemite particles)
and ultrasonification of the solution for 30 min. Subsequently, the
PVP solution and the nanoparticle dispersion were mixed by stirring
(600 rpm). The reaction mixture was subsequently stirred for 24 h
at room temperature. The dispersion of the PVP-coated nanoparticle
was not further purified.
After that, these nanoparticle dispersion was added quickly under
stirring (600 rpm) to a dispersion of amino-functionalized silica
particles in ethanol (c ) 0.5 g/L). The mixture was then stirred for
24 h at room temperature. The amount of nanoparticles was calculated
to be sufficient to provide a 2.5 monolayer coverage of nanoparticles
on the silica colloids. The resulting particles were centrifuged at 20g
(larger colloids) or 400g (smallest colloids), and the supernatant
was removed. In this way, the colloids were separated from the
excess of unadsorbed nanoparticles. The sediments were redispersed
in a solution of 4.2 vol % of ammonia (29.3 wt % NH3 in water)
in ethanol. Immediately after this, TES was added under stirring
(600 rpm) in four steps over 5 h (in the case of QD) or at once (in
the case of gold and maghemite particles). In a typical reaction, in
total 93 µL of TES were added to a dispersion of 36 mg of QD-
decorated silica colloids (35 nm radius) in 105 mL of solvent mixture.
The reaction mixture was then stirred for another 12 h. After that,
the multicore colloids were purified by repeated centrifugation (at
80g (large colloids) or 400g (small colloids)) and redispersion of the
sediment in ethanol. The obtained colloidal particles are soluble in
various polar solvents, such as water, various alcohols, and dimethyl
sulfoxide.
Probing the Stability of QD-Doped Multicore Colloids. A
dispersion of QD-doped multicore colloids in PBS (phosphate-
buffered saline, pH ) 7.4) was kept for 7 days at 37 °C in order to
measure their stability under physiological conditions.
The colloids were exposed to ultraviolet radiation, where a
dispersion of QD-doped multicore colloids in PBS was placed under
a 16 W UV lamp (NU-8-KL from Benda) at 365 nm wavelength.
The UV lamp had a distance to the samples of ∼10 cm. The exposure
time was varied between 1 and 8 h.
All samples were exposed to air for at least 3 days before and also
during the experiment.
The samples were centrifuged at least two times at 400g to remove
the colloids before the ICP-MS analysis that is used to determine
the Cd2+ concentration. The supernatant was used for the measure-
ments.
Characterization. Samples for transmission electron microscopy
(TEM) were prepared by dipping 400 mesh copper grids coated by
a thin carbon film (Pacific Grid-Tech) into the dispersions containing
the particles. The measurements were carried out by using a Zeiss
EM 10 transmission electron microscope operated at 80 keV. The
size of the particles was determined by the measurement tools of
the software Simple PCI (C-Images) by binarization of the images,
separation of the particles by standard image analysis techniques,
(36) Massart, R. IEEE Trans. Magn. 1981, 17, 1247.
(37) van Ewijk, G. A.; Vroege, G. J.; Philipse, A. P. J. Magn. Magn. Mater.
1999, 201, 31.
(38) Arriagada, F. J.; Osseo-Asare, K. J. Colloid Interface Sci. 1999, 211, 210.
(39) van Blaaderen, A.; Vrij, A. J. Colloid Interface Sci. 1993, 156, 1.
(40) Waddell, T. G.; Leyden, D. E.; DeBello, M. T. J. Am. Chem. Soc. 1981,
103, 5303.
Graf et al.
and an estimation of the mean particle radius from the measured
surface area. A diffraction grating (2160 lines/mm) was used to
calibrate the magnification.
Scanning electron microscopy (SEM) was used to study the
formation of ordered structures of the multicore silica colloids. The
samples were prepared by controlled vertical drying of an ethanol
dispersion containing multicore silica colloids on a glass microscope
slide.41 A thin layer of palladium-gold alloy is sputtered on the
samples prior to imaging. The measurements were carried out using
a Zeiss DSM 962 scanning electron microscope.
The UV/visible extinction spectra of free nanoparticles were
measured from dilute aqueous (gold nanoparticles) or chloroform
solutions (CdSe/ZnS nanoparticles) in 1.00 cm quartz cells using a
Perkin-Elmer LAMBDA 19 UV-vis-NIR spectrometer. The spectra
of the nanoparticles embedded in silica colloids were measured in
a mixture of DMSO and ethanol (3.13:1, QD), respectively, DMSO
and water (4.13:1, for studies on gold nanoparticles) to match the
refractive index of silica (nD ) 1.45).
Photoluminescence spectra of the free and the embedded CdSe/
ZnS nanoparticles were measured from the same samples that were
studied by extinction spectroscopy. We used an Aminco Bowman
Series 2 luminescence spectrometer. The excitation wavelength was
458 nm. The quantum yields were determined using Coumarin 6 in
ethanol (QY ) 0.89; λEX ) 458 nm) as a calibration standard.42
About 100 µL of a dilute aqueous dispersion was spread and dried
on a microscope cover slip of 0.15 mm thickness and 22 mm diameter
in order to obtain confocal micrographs of the semiconductor
nanoparticle-doped silica particles. The measurements were carried
out in the fluorescence mode with a Leica inverted microscope (DM
IRB) equipped with a Leica confocal scanning unit (TCS NT) and
a combined krypton/argon mixed gas laser (λ ) 488 nm). We used
a 100× oil immersion objective lens (Leica) with a numerical aperture
of 1.4. The fluorescent light emitted from the particles was detected
above 505 nm.
1H NMR spectra were measured by using a Bruker AC 250 FT-
NMR (250 MHz) spectrometer at 298 K.
The inductively coupled plasma-mass spectrometry (ICP-MS)
measurements were carried out using a Varian ICP-MS spectrometer.
As a calibration standard, an ICP multielement standard solution
from Merck (OC476565) containing 10 mg Cd2+/L was used.
Results and Discussion
The controlled embedding of nanoparticles into silica colloids
requires as a first step the functionalization of the nanoparticles
that enables their attachment on the silica surface in an appropriate
solvent. In particular, high-quality CdSe/ZnS nanoparticles, which
are usually obtained from organometallic syntheses, are stabilized
by organic ligands that render them hydrophobic. On the other
hand, silica colloids are usually dispersed in polar solvents. Several
methods for a hydrophilic surface functionalization of hydro-
phobically stabilized semiconductor nanoparticles have been
reported in the literature.11,43,44 These methods are based on an
exchange of the apolar ligands against more polar ones. However,
such exchange reactions are in general specific to the surface
composition of the nanoparticles, and therefore, they cannot be
simply transferred from one system to another. We did not succeed
to apply successfully methods for hydrophilic functionalization
described in refs 11, 43, and 44 on TOPO-functionalized CdSe/
ZnS nanoparticles prepared by a variant of synthesis routes
described in refs 32 and 33 (see Supporting Information).
Probably, small changes in the QD composition and surface
structure already change the ligand exchange behavior signifi-
(41) Jiang, P.; Bertone, J. F.; Hwang, K. S.; Colvin, V. L. Chem. Mater. 1999,
11, 2132.
(42) Fletcher, A. N.; Bliss, D. E. Appl. Phys. 1978, 16, 289
(43) Hiramatsu, H.; Osterloh, F. E. Langmuir 2003, 19, 7003
(44) Wargnier, R.; Baranov, A. V.; Maslov, V. G.; Stsiapura, V.; Artemyev,
M.; Pluot, M.; Sukhanova, A.; Nabiev, I. Nano Lett. 2004, 4, 451
Embedding of Nanoparticles in Silica Coloids
Figure 2. 1H NMR spectrum of TOPO-stabilized CdSe/ZnS
nanoparticles (a), pure PVP-10 (b), and PVP-stabilized CdSe/ZnS
nanoparticles (c) in CDCl3.
cantly, and so it becomes impossible to apply known function-
alization methods from one system to a closely related one.
Therefore, we used an alternative way based on an adsorption
of the amphiphilic polymer PVP on the nanoparticles. PVP (see
Figure 1) is an amphiphilic polymer that is soluble in water and
many nonaqueous solvents.45,46 This behavior arises from the
presence of a highly polar amide group within the pyrrolidone
ring and apolar methylene and methine groups in the ring and
along its backbone.47 Due to its amphiphilic character, it can be
simply adsorbed on many different surfaces, especially on various
colloidal particles such as metal, oxide, and polymer particles.22
Further, it has been shown recently that a smooth silica shell can
be directly grown on PVP-stabilized particles.22 This facilitates
the growth of the protective layer in the next reaction step (see
below). We have successfully used this PVP-coating procedure
for gold nanoparticles and for the first time for hydrophobic
(TOPO/TOP)-stabilized semiconductor, as well as hydrophilic
iron oxide, nanoparticles. Subsequently, the particles were stable
in alcohols where the adsorption reaction takes place.
The silica particles were functionalized with APS;39 this permits
the attachment of the nanoparticles on silica. APS is used in the
literature as a silane coupling agent for various particles and
surfaces, such as gold10 and silver48 nanoparticles, platinum and
nickel surfaces, hydrophilic stabilized CdTe,49 ZnSe,49 and CdS50
quantum dots, or iron oxide particles.51 The PVP-functionalized
particles can then be simply adsorbed on the amino-functionalized
colloids. Test reactions between APS and PVP show that PVP
does not react with the amino groups but acts only as a stabilizer
in this reaction step, as evidenced from 1H NMR measurements.
1H NMR measurements were also carried out to study the
adsorption reaction between PVP and the TOPO-stabilized
nanoparticles (see Figure 2). The spectra of PVP-stabilized CdSe/
ZnS nanoparticles show after careful cleaning by centrifugation
(43 000g) and redispersion in chloroform (see Figure 2c) that
both the original stabilizer TOPO (see Figure 2a) and PVP (see
Figure 2b) are present. This prooves that after the adsorption
step both compounds are attached to the nanoparticles. Fur-
thermore, it is possible to remove the PVP from the quantum dot
(45) Molyneux, P. Water-soluble synthetic polymers: properties and behaViour;
CRC Press, Inc.: Boca Raton, FL, 1983.
(46) Franks, F. Water: A ComprehensiVe Treatise; Plenum Press: New York,
1982; Vol. 3.
(47) Smith, J. N.; Meadows, J.; Williams, P. A. Langmuir 1996, 12, 3773.
(48) Pastoriza-Santos, I.; Liz-Marzan, L. M. Langmuir 1999, 15, 948.
(49) Yang, P.; Li, C. L.; Murase, N. Langmuir 2005, 21, 8913.
(50) Hebalkar, N.; Kharrazi, S.; Ethiraj, A. S.; Urban, J.; Fink, R.; Kulkarni,
S. K. J. Colloid Interface Sci. 2004, 278, 107.
(51) Wang, F. H.; Yoshitake, T.; Kim, D. K.; Muhammed, M.; Bjelke, O.;
Kehr, J. J. Nanopart. Res. 2003, 5, 137.
Langmuir, Vol. 22, No. 13, 2006 5607
Figure 3. TEM images of PVP-functionalized nanoparticles
adsorbed on silica colloids: CdSe/ZnS nanoparticles (a), gold
nanoparticles (b), and maghemite nanoparticles (c). TEM images of
the same samples after the growth of a thin outer silica layer: CdSe/
ZnS nanoparticles (d), gold nanoparticles (e), and maghemite
nanoparticles (f). In a second growth step, a thicker silica shell was
grown on the CdSe/ZnS multicore silica colloids (g).
surface simply by shaking out a chloroform or butanol dispersion
of CdSe/ZnS quantum dots with water. Subsequently, the original
TOPO-stabilized quantum dots are regained. Therefore, it can
be concluded that PVP does not exchange with the stabilizer
groups on the particles surface. It is rather reversibly adsorbed
on the particle. Probably, the PVP forms a loose network around
the nanoparticles that is on one hand stable enough to prevent
the nanoparticles from aggregation in the particular solvent. On
the other hand, it is still penetrable enough to allow a reaction
between the nanoparticles and the aminogroups on the colloidal
surface.
The adsorption of the nanoparticles on the silica colloids and
their coverage by an outer silica coating leads to embedded
nanoparticles. These were studied by TEM (Figure 3). The
nanoparticles are evenly distributed on the silica particle surfaces
(see Figure 3a-c). However, in a similar approach reported by
Pastoris-Santos et al.,20 citrate-functionalized gold particles are
directly attached to silica particles without a stabilizing polymer.
In this case, only an inhomogeneous distribution in the coverage
density per sphere was reached.
Aggregation of nanoparticles is not observed. The surface
concentration on the silica colloids, and with this the interparticle
distance between the nanoparticles, is adjusted by the number
of nanoparticles-to-surface ratio. The upper limit of surface
coverage is about 30% (see also the optical characterization
below), which is due to Coulomb repulsion between the charged
nanoparticles.52 It is also possible to attach larger nanoparticles,
5608 Langmuir, Vol. 22, No. 13, 2006
Figure 4. Scanning electron microscope image of a piece of a
colloidal crystal of silica colloids (200 nm radius) with an embedded
layer of CdSe/ZnS nanoparticles (1.6 nm radius) prepared by
controlled vertical drying.
such as gold nanoparticles of 20 nm radius in the same way on
silica colloids of 188 nm radius. However, with decreasing
nanoparticle-to-matrix-colloid ratio this adsorption process
becomes significantly less effective.
The nanoparticle-functionalized colloids are directly coated
by a Stöber-like growth reaction yielding a smooth silica shell
(see Figure 3d-f). In the case of the maghemite-doped colloids
shown in Figure 3f, the excess of unadsorbed nanoparticles was
not completely removed before the silica growth step. Therefore,
single silica-coated maghemite nanoparticles were grown on these
colloids. In the other two cases (Figure 3d, e), this was prevented
by thoroughly cleaning the samples before the shell growth. The
thickness of the outer shell was controlled by the added amount
of TES. It is also possible to grow thicker silica layers by several
consecutive growth steps (see Figure 3g). The size polydispersity
of the final nanoparticle-doped silica colloids decreases with
increasing silica shell thickness. For a 4 nm thick shell (Figure
3d), the polydispersity is 4.1%, and for a 14 nm thick shell
(Figure 3g), the polydispersity is 2.3%. Similar results have been
reported before for the seeded growth of pure silica particles.30
This outer silica shell opens a variety of possible surface
modifications24,39 and also the option to attach a second layer of
nanoparticles on the colloids in a controlled distance from the
first layer.
As the attached nanoparticles are small compared to the total
particle size and as the outer silica layer grows smoothly and
homogeneously on the nanoparticle-functionalized colloids, the
polydispersity of the final colloids is mainly determined by the
particle size distribution of the primary silica cores. Silica cores
can easily be prepared with low polydispersity by standard
methods (see above) so that the overall polydispersity of the
final particles is also low (2-5%). As a result, the particles
crystallize easily into highly ordered structures, as is known for
silica colloids.53,54 This is also facilitated by the closed outer
silica layer. Figure 4 shows a colloidal crystal of silica colloids
(200 nm radius) with an embedded layer of CdSe/ZnS nano-
particles (1.6 nm radius) prepared by controlled vertical drying41
of an ethanol dispersion on a glass microscope slide.
The optical properties of the nanoparticles before and after the
adsorption and embedding steps were investigated by extinction
and photoluminescence spectroscopy. The spectra of the multicore
particles were measured in solvent mixtures (water/DMSO or
ethanol/DMSO) which match the refractive index of silica
(nD ) 1.45 at 25 °C). Because of their different solubility
(52) Westcott, S. L.; Oldenburg, S. J.; Lee, T. R.; Halas, N. J. Langmuir 1998,
14, 5396.
(53) Denkov, N. D.; Velev, O. D.; Kralchevsky, P. A.; Ivanov, I. B.; Yoshumura,
H.; Nagayama, K. Langmuir 1992, 8, 3183.
(54) Denkov, N. D.; Velev, O. D.; Kralchevsky, P. A.; Ivanov, I. B.; Yoshumura,
H.; Nagayama, K. Nature 1993, 361, 26.
Graf et al.
properties, the spectra of the free citrate-stabilized gold particles
are measured in water (nD ) 1.33 at 25 °C) and those of the
TOPO-stabilized CdSe/ZnS particles are measured in chloroform
(nD ) 1.44 at 25 °C).
At a low degree of surface coverage, the extinction spectra of
gold nanoparticles embedded in the silica colloids are nearly
unchanged compared to the free particles. At a higher degree of
coverage (see TEM micrograph, Figure 3e), the extinction spectra
of the adsorbed and embedded particles are slightly red-shifted
by 18 and 10 nm, respectively, compared to the free gold
nanoparticles (Figure 5a, solid line, λmax ) 522 nm). This shift
is larger for the adsorbed particles (Figure 5a, dashed line,
λmax ) 540) than for the embedded ones (Figure 5a, dotted line,
λmax ) 532 nm). Remarkably, the shape of the spectra is not
changing. Due to the change of the refractive index from water
(nD ) 1.33) to silica (nD ) 1.45 at 25 °C) and PVP (nD ) 1.46
at 25 °C) a red-shift of about 5 nm is expected, according to
calculations based on Mie theory (see Supporting Information).
The slightly larger shift for the embedded particles can be
explained by considering dipole-dipole interactions between
the gold nanoparticles, which lead to red-shifts in the absorption
cross section.20 The fact that this change is still small at high
nanoparticle concentrations and that the shape of the spectra
remains unchanged shows that clustering and aggregation of
nanoparticles can effectively be avoided by PVP coating. If the
adsorbed particles are not capped with an outer silica layer, they
are likely mobile on the surface, which leads to increased
interactions and a larger spectral shift.
In the case of the CdSe/ZnS nanoparticles, the extinction
spectrum remains nearly unchanged before and after embedding
them in silica colloids (Figure 5b). Specifically, the peak at 562
nm does not shift after embedding. Therefore, the intensity of
the peak at 562 nm can also be used to estimate the quantum dot
concentration in the silica colloids, assuming that the silica matrix
does not contribute to the extinction because of the index-matching
solvent.55 An average concentration of 300 semiconductor
nanoparticles per silica colloid was found. This corresponds to
a surface coverage of about 16% after the adsorption step. For
larger silica colloids of 188 nm core radius, an average
concentration of 16 000 semiconductor nanoparticles per silica
colloid was derived, corresponding to a surface coverage of about
29% after the adsorption step.
The photoluminescence spectra remain unchanged after
adsorption of the polymer as well as after attaching of the PVP-
stabilized QD to the silica spheres. However, after the growth
of the outer silica shell, the emission maximum is slightly red-
shifted (from 581 to 587 nm, see Figure 5c). The width of the
spectra is not changing during all reaction steps. This shows that
the polydispersity of the QD is not changing. Unlike these findings,
other authors reported a significant blue-shift of the emission
spectra after the functionalization of QD with silica coupling
agents and after their adsorption or embedding in a silica shell.21,56
These blue-shifts can be described to a decrease of the size of
the semiconductor core due to a partial oxidation of the nanocrystal
surface during exchange of the stabilizer and during the ripening
of the silica layer.56 Evidently, such a decrease in size can be
avoided by the present approach.
The photoluminescence quantum yield (QY) remains nearly
unchanged before and after absorption of PVP (e.g., it changes
from 21% to 19%). However, the adsorption on the amino-
(55) Schmelz, O.; Mews, A.; Basché, T.; Herrmann, A.; Müllen, K. Langmuir
2001, 17, 2861.
(56) Rogach, A. L.; Nagesha, D.; Ostrander, J. W.; Giersig, M.; Kotov, N. A.
Chem. Mater. 2000, 12, 2676.
Embedding of Nanoparticles in Silica Coloids
Figure 5. Extinction spectra of citrate-stabilized gold nanoparticles
(9 nm radius) in ethanol (solid line) and silica colloids (192 nm
radius) with adsorbed gold nanoparticles before (dashed line) and
after the growth (dotted line) of an outer silica shell measured in
DMSO/water (nD ) 1.45) (a). Extinction (b) and photoluminescence
(c) spectra of TOPO-stabilized CdSe/ZnS nanoparticles (1.6 nm
radius) in chloroform (solid lines) and PVP-stabilized CdSe/ZnS
nanoparticles embedded in silica colloids (40 nm radius) in DMSO/
ethanol (nD ) 1.45) (dotted lines). Confocal microscopy image
(fluorescence mode, excitation wavelength 488 nm) of silica colloids
with an embedded layer of CdSe/ZnS nanoparticles (total radius 40
nm) (d).
Langmuir, Vol. 22, No. 13, 2006 5609
functionalized silica colloids approximately halves the QY to ca.
10% and further growth of an outer silica reduces the QY to ca.
1%. As already concluded from NMR results, likely the PVP is
only attached to the TOPO-stabilized particles and this adsorption
step only involves the organic periphery of the QD. Therefore,
it does not alter the properties of the inorganic core. In the next
step, the semiconductor nanoparticles react with the amino groups
on the silica sphere. Presumably, this reaction occurs on the
surface of the ZnS/CdSe core so that a change in the luminescence
properties is observed. The more significant decrease of the QY
during the growth of the silica shell is probably due to water and
ammonia present in the Stöber growth process, which leads to
the creation of more severe surface defects on the QD. These are
likely caused by oxidation reactions. The fact that, despite PVP
coating, this last reaction step, as well as the adsorption of the
nanoparticles on the silica colloids, strongly influences the optical
properties of the CdSe/ZnS nanoparticles gives evidence that the
PVP forms only a loose network around the nanoparticles. This
network is sufficient to stabilize of the QD in more polar solvents.
However, it is penetrable for small ions and molecules.
Similar or much lower QY have been reported for other coating
or embedding processes with QD.21,56 However, it should be
considered that most of these reactions require the use of water-
soluble QD, which cannot be produced with an as high QY as
the apolar-stabilized QD from organometallic syntheses, so that
in these cases, the final QY of the embedded particles is
considerably lower.
The fluorescence yield per nanoparticle-doped silica colloid
is also high, which is due to the substantial QD content per silica
sphere. Measurements using confocal laser scanning microscopy
demonstrate that the single multicore colloids show a bright
fluorescence (see Figure 5d). Furthermore, these particles have
a much higher photostability if they are excited by light in the
visible regime compared to colloidal particles labeled with
conventional organic dye molecules, such as rhodamine57 or
fluoresceine.58 For instance, after 30 min of bleaching with a Hg
lamp (50 W) at 546 nm, the fluorescence intensity of these
colloidal particles remains unchanged, whereas under the same
conditions, silica colloids labeled with rhodamine significantly
bleach already within 20 s.
We have also investigated the stability of the QD-doped
particles against leakage of toxic ions into the solution. One of
the main problems related with the use of QD as markers in life
science is their cytotoxicity. For CdSe-containing QD, this is
mainly caused by the release of free Cd2+ ions due to deterioration
of the CdSe lattice through oxidation processes. The cytotoxicity
of QD is enhanced by exposure to ultraviolet light and to air.9
In contrast, the successive addition of inorganic layers, such as
ZnS, small organic ligands, or a cross-linked organic shell
progressively minimizes the cytotoxicity of QD, so that the
biocompatibility of CdSe QD is substantially increased.9 The
amount of free Cd2+ can be correlated with the degree of observed
cytotoxicity, as shown in ref 9. The exposure to 100-400 µM
(11-44 ppm) Cd2+ leads to significant cell death.59 Some cell
types, such as cardiomyocytes, are sensitive to even lower levels
of Cd2+ exposure (0.1 µM or 11 ppb), whereas other cell types
are more resistant.60
To probe the stability of the QD-doped multicore particles
dispersion of the particles, these were kept for 7 days under
physiological conditions (37 °C, PBS with pH ) 7.4). Further,
(57) Verhaegh, N. A. M.; van Blaaderen, A. Langmuir 1994, 10, 1427.
(58) van Blaaderen, A.; Vrij, A. Langmuir 1992, 8, 2921.
(59) Santone, K. S.; Acosta, D.; Bruckner, J. V. J. J. Toxicol. EnViron. Health
1982, 10, 169.
(60) Limaye, D. A.; Shaikh, Z. A. Toxicol. Appl. Pharm. 1999, 154, 59.
5610 Langmuir, Vol. 22, No. 13, 2006
dispersions of the multicore particles were exposed to a UV light
source (16 W) at 365 nm for 1-8 h. All samples were exposed
to air for at least several days before and also during the
experiments. Subsequently, the samples were centrifuged to
remove the multicore particles, and the remaining solutions were
tested for free Cd2+ by ICP-MS. We observe for particles with
a silica shell of <5 nm thickness and a colloidal particle
concentration corresponding to 1 g QD/L, a Cd2+ concentration
of 54 ( 2 ppb after 7 days. This is considerably lower than
reported earlier, where concentrations of about 1 ppm for c )
0.25 g QD/L were reported for ZnS/CdSe QD, which were only
capped with organic molecules (mercaptoacetic acid, MAA) under
similar conditions.9 Already, these QD were reported to be
virtually not cytotoxic due to oxidation by air during processing.9
In the present case, the exposure to UV light has only a minor
influence on the release of Cd2+. The Cd2+ concentration was
0.74 ( 0.2 ppb after 1 h and increased to 3.2 ( 0.2 ppb after
8 h. This is much lower than reported for MAA-stabilized ZnS/
CdSe QD (23 ppm), bovine serum albumin (BSA)-coated dots
(20 ppm), and for commercial polyacrylate/streptavidin-coated
ZnS/CdSe QD (Quantum Dot Corporation) (15 ppm) recalculated
for similar conditions.9 Obviously, the silica layer efficiently
protects the CdSe core from oxidation processes, including photo-
oxidation and hinders the transport of Cd2+ from the QD into
the dispersion. The small amount of Cd2+, which is still found
in the dispersion, is very likely due to the fact that silica coatings
prepared by Stöber-like growth processes have a microporosity
of 10-15%.61 As a result, they are still penetrable for small
molecules and ions if the covering layers are relatively thin.
However, the diffusion coefficient of oxygen in silica particles
is 4 orders of magnitudes smaller than in water, as shown in ref
62, so that the release of Cd2+ into the solvent is not completely
hindered, but significantly reduced. Further, it should be noted
that, e.g., for typical in vitro cell imaging studies, such high
concentrations, as used in the present experiments, are not
commonly available. Previous work showed that there are 10-
250 times lower concentrations sufficient for imaging studies.63,64
Thus, the present results give an upper Cd2+ concentration limit,
which indicates that Cd-containing QD can be used for cell-
imaging studies. Detailed work on studying the cytotoxicity and
biocompatibility of the particles is in progress.65
Conclusions
A novel method for the controlled embedding of multiple
small particles, such as semiconductor, noble metal, and metal
(61) Bogush, G. H.; Tracy, M. A.; Zukoski, C. F. J. Non-Cryst. Solids 1988,
104.
(62) Li, B.; Xie, Y.; Huang, J. X.; Liu, Y.; Qian, Y. T. Ultrason. Sonochem.
2001, 8, 331.
(63) Wu, X.; Liu, H.; Liu, J.; Haley, K. N.; Treadway, J. A.; Larson, J. P.; Ge,
N.; Peale, F.; Bruchez, M. P. Nat. Biotechnol. 2003, 21, 41.
(64) Jaiswal, J. K.; Mattoussi, H.; Mauro, J. M.; Simon, S. M. Nature Biotechnol.
2003, 21, 47.
(65) Dembski, S.; Graf, C.; Krüger, T.; Ewald, A.; Rühl, E. manuscript in
preparation.
Graf et al.
oxide nanoparticles in silica colloids has been developed.
Nanoparticles that are stable in polar solvents were obtained
from adsorbing PVP on their surface. These particles were
subsequently adsorbed on amino-functionalized silica colloids.
The aggregation of the nanoparticles on the colloidal surface is
prevented, which is a result of the protective polymer layer. The
interparticle distance can be adjusted by the surface-to-bulk ratio
of the nanoparticles. Finally, the nanoparticle-decorated silica
colloids are directly coated by a silica layer of variable thickness
by the addition of tetraethoxysilane (TES) to an ammonia/ethanol
solution containing the particles in a seeded growth process.
Because of the well-controlled interparticle distance, the optical
properties of the single gold nanoparticles are largely kept
unchanged in the silica matrix. In the case of semiconductor
nanoparticles, the shape of the emission spectra remains constant,
whereas the fluorescence quantum yield is reduced. Nevertheless,
the present results from confocal laser scanning microscopy show
that the semiconductor nanocrystal-doped colloids have a bright
fluorescence and they are highly stable against bleaching. Leakage
of toxic ions, such as Cd2+, from the embedded nanocrystals is
strongly reduced under oxidative conditions. Therefore, these
particles are suitable systems for the use as markers in life science
or as model systems for diffusion and phase behavior studies on
colloids by fluorescence microscopy methods. Furthermore, it
is possible to adsorb and embed a second or more layers of
nanoparticles on silica colloids containing an embedded layer of
nanoparticles. This opens the opportunity to obtain a high degree
of labeling and to combine the properties of several types of
nanoparticles in one particle system, e.g., magnetic properties
and strong luminescence. We also showed that it is possible to
form highly ordered colloidal crystals of these nanoparticle-
doped particles. This opens, combined with the controlled
embedding of nanoparticles, the possibility for the fabrication
of novel photonic crystals.
Acknowledgment. The authors gratefully acknowledge Prof.
G. Krohne and Dr. R. Wolf (Biozentrum, Universität Würzburg,
Germany) and for the use of the electron microscopes, Prof. A.
van Blaaderen (Debye Institute, Universiteit Utrecht, The
Netherlands) for the use of the confocal laser scanning microscope,
Dr. M. Grüne (Institut für Organische Chemie, Universität
Würzburg, Germany) for the NMR measurements, and Dr. U.
Gbureck (Lehrstuhl für Funktionswerkstoffe der Medizin und
der Zahnheilkunde des Universitätsklinikums Würzburg, Ger-
many) for the IPC-MS measurements. This work was supported
by the Deutsche Forschungsgemeinschaft (Projekt C8, Sonder-
forschungsbereich 410), the Universität Würzburg, and the Fonds
der Chemischen Industrie.
Supporting Information Available: Synthetic detail and data.
This material is available free of charge via the Internet at http://
pubs.acs.org.
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