View Online / Journal Homepage / Table of Contents for this issue

PAPER www.rsc.org/analyst | Analyst

Poly(ethylene glycol)-stabilized silver nanoparticles for bioanalytical

applications of SERS spectroscopy
Andriy Shkilnyy,a Martin Souce,a Pierre Dubois,a Fabienne Warmont,b Marie-Louise Saboungib
and Igor Chourpa*a
Received 23rd March 2009, Accepted 16th June 2009
First published as an Advance Article on the web 6th July 2009
DOI: 10.1039/b905694g

The present work depicts the efficient one-step synthesis and detailed evaluation of stable aqueous
colloids of silver nanoparticles (NPs) coated with poly(ethylene glycol) (PEG) covalently attached to
their surface. Due to steric repulsion between polymer-modified surfaces, the stability of the
Published on 06 July 2009 on http://pubs.rsc.org | doi:10.1039/B905694G

nanoparticle suspension was preserved even at high ionic strength (0.1 M NaCl). At the same time, the
PEG coating remains sufficiently permeable to allow surface-enhanced Raman scattering (SERS) from
micromolar concentrations of small molecules such as the anticancer drug mitoxantrone (MTX). The
enhancement efficiency of the hot spot-free Ag-PEG was compared to that of citrate-stabilized Ag
Downloaded by University of Sussex on 31 May 2012

colloids used after pre-aggregation. The potential of the polymer-stabilized colloids developed in this
study is discussed in terms of bioanalytical applications of SERS spectroscopy.

Introduction Most of the analytical and bioanalytical applications of SERS

The past decade has been particularly rich in the development of
novel nanoparticle systems with innovative composition aiming
to provide designed properties. This spectacular progress in
nanotechnology logically inspires the research community trying
to push the frontiers of analytical methods. Surface-enhanced
Raman scattering (SERS) spectroscopy is one of the methods
that are most strongly concerned by advanced nanoparticle (NP)
design.1–5 In particular, SERS relies on NPs made of noble metals
(Au, Ag, Pt) that have localised surface plasmonic resonance
(LSPR). Compared to conventional Raman spectroscopy, the
enhancement of Raman scattering on Ag and Au NPs provides
a sensitivity gain of typically about six orders of magnitude,
while for single-molecule SERS experiments enhancement
factors of 1014–1015 were reported.4,6 The efficiency of spectro-
scopic detection of analytes by nanoparticle-based SERS is
known to depend on three major considerations: (i) the Raman
activity of the analyte, (ii) the LSPR quality of the nanoparticle
and (iii) last but not least – the surface chemistry phenomena.
Indeed, one of the crucial requirements for the enhancement of
Raman scattering is that the analyte molecules come close to the
vicinity of the metal surface. For a given analyte molecule, the
adsorption quantity and quality (geometry) is dependent on the
affinity analyte–metal, which typically involves electrostatic or
chemical interactions. Therefore, the SERS signal depends on the
final equilibrium between the analyte molecules retained on the
particle surface and those remaining in the surrounding solution.
a polymer chains in solution which sterically hindered the forma-
EA 4244, Physicochimie des mat eriaux et des biomol ecules, groupe
Nanovecteurs magn etiques pour la chimioth erapie, Universit
e F.
Rabelais, Faculte de Pharmacie, 31 avenue Monge, F-37200 Tours,
France. E-mail: igor.chourpa@univ-tours.fr; Fax: +33 2 47 36 72 70;
Tel: + 33 2 47 36 71 62
b
UMR 6619, Centre de Recherche sur la Mati ere Divis
ee, CNRS,
Universit
e d’Orl
eans, 1B rue de la F
erollerie, F-45071 Orl
eans cedex 2,
France
spectroscopy involve aqueous suspensions of Ag and Au NPs,
typically prepared by soft chemistry methods. The stability of
those suspensions is assured by electrostatic repulsion of nano-
particles carrying counter-ions like citrates.7 With those particles,
the pre-analytical step includes particle aggregation, because the
interparticle junctions (named hot spots) are particularly favor-
able for enhancing Raman scattering.8 This aggregation involves
destabilizing the particle solvation layers and can be achieved
either by increasing the ionic strength of the medium or directly
by the adsorption of certain analytes.9,10 Unfortunately, the
destabilized NPs aggregate in a non-controlled manner and
precipitate quite rapidly. This is particularly problematic for
bioanalytical applications of SERS spectroscopy, like those
made with live cells.11 Indeed, the composition of most biologi-
cally relevant media provokes rapid aggregation and precipita-
tion of electrostatically-stabilized silver and gold colloids.5 In
addition, the transport of aggregated NPs through the cell
membrane and their distribution in subcellular compartments is
more difficult.
Coating the NP surface with layers of neutral organic mole-
cules can efficiently protect them from aggregation by steric
repulsion.12 There have been numerous studies on the effect of
poly(ethylene glycol) (PEG) on silver NP stability.13–16 For
instance, Luo et al.14 investigated the reduction of silver nitrate in
the presence of PEG. The authors showed that increasing the
molecular weight of the polymer facilitates the formation of
silver NPs, and the obtained colloids were stable for months. The
stabilization effect was achieved due to the presence of the free
tion of aggregates.
In the present paper, we designed and evaluated stable
aqueous colloids of Ag NPs intended to overcome the limitations
of the conventional, electrostatically-stabilized NPs. We present
our own method for the preparation of silver NPs with PEG
chains covalently attached to the surface (Ag-PEG). Such

1868 | Analyst, 2009, 134, 1868–1872 This journal is ª The Royal Society of Chemistry 2009

a system would permit a higher colloid stability without a big
excess of the polymer in solution.12 If the density of covalently
bound polymer groups is not too high, part of the surface
remains available for the adsorption and the SERS detection of
analytes.3,17
Ag-PEG nanoparticles were obtained by an original one-step
synthesis protocol in which the just-formed silver NPs directly
react with the thiol group of PEG. In this manner, we obtain fully
functionalized and nearly monodisperse silver NPs with good
yield. The particles were characterized in terms of morphology,
size and colloidal stability, in comparison to conventional
Ag-citrate sols. The stability of the suspension was confirmed by
DLS (dynamic light scattering) and UV-visible spectroscopy.
Adsorption of chloride ions and the anticancer drug mitoxan-
Published on 06 July 2009 on http://pubs.rsc.org | doi:10.1039/B905694G
trone (MTX) on the PEGylated silver was assessed through the
LSPR absorption band measured in the visible wavelength
range. Finally, the SERS efficiency of the hot spot-free Ag-PEG
and pre-aggregated Ag-citrate NPs was compared.
Downloaded by University of Sussex on 31 May 2012
Experimental
Chemicals
All chemicals and reagents were of analytical grade and used as
received. Silver nitrate (AgNO3), sodium citrate dihydrate,
sodium borohydride (NaBH4), sodium chloride (NaCl), mitox-
antrone (1,4-dihydroxy-5,8-bis[(2-[(2-hydroxyethyl)amino]ethyl)
amino]-9,10-anthracenedione) (MTX) and O-[2-(3-Mercapto-
propionylamino)ethyl]-O0 -methylpolyethylene glycol (PEGSH)
(MW 5000 g/mol) were obtained from Sigma-Aldrich (France).
Ethanol was purchased from Carlo Erba (France). Ultrapure
water was produced using a Barnstead EASYpure RoDi system
(Thermo Fisher Scientific, France). Dialysis membranes
(MWCO 8000 Da) were purchased from Interchim (Montluçon,
France).
Synthesis of nanoparticles
The colloids of PEGylated silver nanoparticles (Ag-PEG) were
prepared in an ethanol solution employing thiol chemistry and
using a method modified from literature.18 The reaction (Fig. 1)
was carried out under nitrogen atmosphere to prevent oxidation
reactions during formation of the metal particles. In a typical
Fig. 1 Schematic of Ag-PEG NP synthesis and incubation with MTX
for SERS spectroscopy experiments.
This journal is ª The Royal Society of Chemistry 2009
View Online
experiment, 4 mL of an ethanolic silver solution (0.0075 M
AgNO3) were mixed with 0.075 g of PEGSH. Upon mixing, the
obtained suspension was sonicated for 5 min. Then, 1.3 mL of
a 0.09 M NaBH4 ethanol solution was added dropwise from
a syringe under vigorous stirring. The obtained suspension was
kept at room temperature for 2 h. Next, 2.5 mL of the dark
brown suspension was diluted with deionized water up to 10 mL
and subsequently dialyzed against water in order to remove
excess polymer.
The Ag-citrate colloids were prepared by reducing silver
nitrate in presence of an excess of trisodium citrate, according to
a standard protocol described elsewhere.7 For SERS experi-
ments, the colloids were pre-aggregated by dilution in phosphate
buffered saline (PBS).
Nanoparticle characterization
Dynamic light scattering and z-potential measurements. DLS
experiments were conducted on a Malvern Autosizer 4700 device
at 25  C. The DLS measurements were carried out at the laser
wavelength 633 nm and a detection angle of 173 . The z-potential
measurements were carried out using a Malvern NanoZ instru-
ment at 25  C. In a typical experiment 60 mL (0.0014 M Ag) of
the dialyzed suspension were added to 10 mL of 0.1 M sodium
chloride solution and followed by sonication for 15 min. Samples
were not filtered before measurements.
UV-vis spectrophotometry. UV-vis spectra were acquired with
an Anthelie Advanced Spectrophotometer (Secomam, France).
The concentrations of silver and MTX throughout all experi-
ments were 8.8 mM and 10 mM respectively.
Electron microscopy. TEM experiments were carried out on
a Philips CM20 transmission electron microscope operated at
200 kV. Samples were deposited on carbon-coated Cu grids from
the aqueous suspension. Excess water was removed by filter
paper. The obtained samples were left to dry under ambient air.
Surface-enhanced Raman spectroscopy. SERS measurements
were carried out using a LabRAM confocal microspectrometer
(Horiba Jobin-Yvon, France) equipped with an air-cooled CCD
detector, a spectrograph (300 mm focal length, dispersion grating
of 1800 grooves per mm) and an Olympus BX-40 microscope
with an automated X–Y–Z scanning stage. Sample irradiation
and collection of SERS spectra were performed through a 50
microscope objective (numerical aperture 0.50; LMPlan Fl,
Olympus, Japan). The samples (a 100 mL drop on a microscope
glass slide) were analyzed in the non-confocal mode of the
spectrometer (confocal hole opened to 800 mm). The MTX SERS
spectra were excited with the 488 nm line of an Ar+ laser (36-
LAP-431-230, Melles-Griot, France) with a power on the
samples of ca. 0.8 mW. No sample photodegradation was
observed under the conditions used. Each spectrum was recorded
as an average of 36 scans (0.1–2 s per scan). The spectra pre-
sented in the figures are averages of at least three independent
measurements.
Both experiment control and following data treatment were
performed using the LABSPEC software package.
Analyst, 2009, 134, 1868–1872 | 1869

Results and discussion
Characterization of the nanoparticles in terms of morphology,
size and colloidal stability
One-step synthesis of Ag-PEG NPs (Fig. 1) results in a roughly
spherical morphology as seen in TEM (Fig. 2A). The TEM
images reveal also the presence of two populations of silver NPs
with diameters of ca. 10–20 nm and 1–5 nm. It is noteworthy that
the NPs appeared clearly non-aggregated in TEM. The non-
aggregated state of the Ag-PEG particles was maintained even in
the presence of chloride ions (0.1 M), as was confirmed by DLS
experiments (Fig. 2B). The average distribution of hydrodynamic
diameters (henceforth denoted as HD) exhibited two maxima:
the main one at ca. 70 nm and a minor one at ca. 10 nm. The
Published on 06 July 2009 on http://pubs.rsc.org | doi:10.1039/B905694G
bigger values of HD of particles compared to their size in TEM
correspond to the solvation layers on the surface of silver NPs
composed of organic molecules. As expected for the particles
coated with neutral PEG, the z-potential of the Ag-PEG samples
Downloaded by University of Sussex on 31 May 2012
was close to zero: ca. 3 mV.12 This value confirmed that the
particles are stabilized by steric hindrance and not by ionic
repulsion.
Despite the fact that DLS data for the Ag-citrate colloids also
showed two populations with quite similar HD maxima at ca.
80 and 9 nm, the size distribution peak for Ag-PEG was signif-
icantly narrower. Whereas Ag-PEG colloids were stable in the
Fig. 2 (A) TEM image of Ag-PEG NPs; (B) DLS data on the colloidal
stability of the Ag-PEG NPs compared to the Ag-citrate NPs (see text for
details).
1870 | Analyst, 2009, 134, 1868–1872
View Online
presence or chloride ions, Ag-citrate colloids were not, and their
HD maxima shifted to ca. 220 and ca. 30 nm, respectively. This
illustrates the well-known fact that chloride anions perturb the
citrate protective layer and thus lead to particle assembly with
bigger aggregates.9,10
Hence, the better quality of the Ag-PEG samples over
Ag-citrate ones is expressed in terms of a more narrow size
distribution and of enhanced colloidal stability due to steric
repulsion of PEGylated NPs. This neutral polymer layer should
not only protect the suspensions from the effect of salt concen-
tration but also from moderate fluctuations of pH.12
Accessibility of the PEGylated silver surface for adsorption of
chloride ions and of MTX as seen with UV-vis spectroscopy
Both the Ag-citrate and Ag-PEG NPs in their respective aqueous
suspensions had typical LSPR absorption maxima at 404 nm
(Fig. 3, solid lines). According to the short-wavelength position
of the LSPR band, our silver particles appear to be smaller than
those reported recently in the literature on PEGylated silver NPs
(absorption maxima at 420 nm).19 As expected for NPs with
narrow size distribution, the full width at half-maximum (fwhm)
of the LSPR band was quite small: 77 and 85 nm for Ag-citrate
and Ag-PEG NPs, respectively.20
In addition, the absorption band of Ag-PEG was more
symmetric and decreased to zero absorbance at higher wave-
length, while the Ag-citrate spectrum exhibited a weak, nearly
Fig. 3 UV-vis spectrophotometry data on the adsorption of chloride
ions and MTX onto the surface of Ag-citrate and Ag-PEG NPs: LSPR
band of the colloids before (—) and after addition of chloride ions (-- --);
absorption spectra of MTX and of colloids incubated with MTX alone
(-- . -) and in presence of chloride ions (/). The spectra were normalized
for comparison.
This journal is ª The Royal Society of Chemistry 2009

constant background observable from 650 to 800 nm. This
observation indicates that, in contrast to Ag-PEG, Ag-citrate
samples may contain a higher fraction of larger particles, which
in its turn correlates well with our DLS data discussed above.
In spite of the initial similarity, the two kinds of colloids reacted
differently to the presence of Cl (dashed lines in Fig. 3). After an
addition of 0.1 M NaCl, the Ag-citrate colloids aggregated, and
the interparticle plasmon coupling led to a decrease of the initial
LSPR band and to an increase of background above 500 nm. This
kind of behaviour is typical for Ag-citrate colloids.10 On the other
hand, the LSPR band of the Ag-PEG NPs exposed to Cl did not
decrease and did not give rise to any background at higher
wavelength. Instead, the LSPR band became narrower (78 nm
instead of 85 nm) and blue-shifted (401 nm instead of 404 nm),
Published on 06 July 2009 on http://pubs.rsc.org | doi:10.1039/B905694G
which confirmed that Cl ions have diffused to the silver surface
but the size distribution of the Ag-PEG nanoparticles remained
uniform and well dispersed. A similar effect was described by
Chumanov and co-workers for the carbon-coated silver nano-
Downloaded by University of Sussex on 31 May 2012
particles.21 Therefore, the effect of Cl confirms that the PEG
coating is permeable for small anions and allows them to bind to
the silver surface. On the other hand, the PEG coating is dense
enough to protect the particles from aggregation.
To check if this coating still permitted molecular species to
access the silver surface, we incubated the Ag-PEG NPs in
aqueous solution of the anticancer drug MTX (MW 517.4). After
an incubation for 10–30 min (the time used to prepare samples
for SERS) in an aqueous solution of 10 mM MTX, the plasmon
band of silver was partially decreased (dashed-dotted lines in
Fig. 3) for both types of colloids. In addition, the drug absorp-
tion bands were observed at 609 nm and 660 nm. These results
correspond to the MTX adsorption on the silver surface (Fig. 1).
The effect of MTX adsorption on the plasmonic band decrease
was even more pronounced after 24 h for Ag-PEG, but still no
particle aggregation was observed under 100 objective of
optical microscope (data not shown). Another confirmation of
the colloidal stability was the absence of precipitate after ultra-
centrifugation (2350g for 10 min). Unfortunately, it was not
possible to make DLS measurements on the Ag-PEG-MTX
particles with MTX, because of the strong drug fluorescence
induced by the laser used in the DLS setup.
Since Cl ions are known to influence the adsorption of certain
analytes on silver surfaces,5,10 we investigated the effect of these
ions on our colloids in the presence of MTX. The addition of 0.1 M
NaCl induced a slight red shift of the two absorption maxima of
MTX (to 613 nm and 666 nm) alone or in presence of the two kinds
of colloids (Fig. 3). This red shift and the decrease in intensity of
the high-wavelength band of MTX is assignable to p–p stacking
occurring for the drug molecules in PBS, as described previously
in the literature.22 For both types of colloids incubated with MTX,
the decrease of the LSPR bands was more pronounced in the
presence of Cl than in the absence of the ion. Therefore, the
aggregates of MTX appear to adsorb more efficiently to the silver
surface. Interestingly, the effect of Cl was more pronounced on
the plasmon band of Ag-PEG than on that of Ag-citrates.
SERS activity of Ag-PEG compared to Ag-citrate
We checked the SERS activity of the colloids with two concen-
trations of MTX, 1 mM and 10 mM. These concentrations
This journal is ª The Royal Society of Chemistry 2009
View Online
correspond to the incubation conditions of cancer cells that allow
the intracellular drug to be followed by fluorescence and SERS
spectroscopy.11
As expected, conventional Ag-citrate colloids pre-aggregated
with Cl and incubated for a few minutes with a 1 mM solution of
the drug provided the very intense SERS spectra (top spectrum in
Fig. 4) characteristic of MTX.23 In fact, this concentration is
several orders of magnitude above the detection limits known for
this system.23,24
The most interesting to discuss is the MTX SERS spectra
obtained with Ag-PEG NPs. In our study, a 10 min incubation of
the Ag-PEG colloids with 1–10 mM MTX was sufficient to detect
SERS of the drug. An additional incubation of 30 min resulted in
a nearly 2-fold increase of the SERS intensity. This confirmed
that the observed enhanced Raman scattering was dependent on
the diffusion of the drug through the PEG coating.
The SERS data confirmed that MTX can be adsorbed on the
silver surface of PEGylated nanoparticles, both in the absence
and in the presence of Cl ions. Nevertheless, the chloride anions
resulted in a moderately increased SERS intensity (about
15–20%). In addition, the MTX SERS spectra recorded on the
Ag-PEG-Cl nanoparticles exhibited an increased band ratio at
1177/1201 and 1444/1413 cm1 and a less resolved doublet at
1500–1526 cm1 (Fig. 4). These changes in the presence of
chloride anions may be assigned to the different adsorption
geometry and/or to the partial aggregation of MTX described
above. Finally, the MTX spectra from the Ag-PEG-Cl were
of very similar shape compared to those from the non-coated
Ag-Cl.
The maximal average intensity of the MTX spectra from the
Ag-PEG-Cl nanoparticles was 130 times lower than that
obtained with Ag-Cl (compared after intensity normalizing on
the OH stretching band of water at 3400 cm1). In addition to the
limited drug diffusion through the polymer, this intensity loss
may be related to the reduced enhancement in the absence of hot
Fig. 4 SERS spectra of MTX on Ag-citrate and Ag-PEG NPs. For
visual comparison, the MTX spectrum from Ag-Cl was reduced
15 times.
Analyst, 2009, 134, 1868–1872 | 1871

spots.9 Nevertheless, even the two orders of magnitude lower
enhancement factor is still interesting for analytical applications.
For instance, a 1 mM MTX solution was still sufficient to obtain
good quality SERS spectra (S/N ratio >10 with 36 scans of 2 s).
Interestingly, a 10-fold decrease in concentration (from 10 to 1
mM MTX) corresponded to an 8.5-fold SERS intensity decrease,
thus indicating that at the higher concentration of the drug we
are approaching saturation. Scan-to-scan random intensity
fluctuations were much less pronounced with the stable Ag-PEG
suspensions than with aggregated Ag-Cl. This stability and
almost linear behaviour might allow better quantitative
measurements with Ag-PEG.
To the best of our knowledge, there are no literature reports on
polymer-modified NPs permitting SERS of analytes diffusing
Published on 06 July 2009 on http://pubs.rsc.org | doi:10.1039/B905694G
from the surrounding solution to the silver surface through the
polymeric layer. For instance, according to the recent report by
Chumanov co-workers,21 the carbon-coated silver NPs procure
no SERS of adenine, although the chloride anions were able to
Downloaded by University of Sussex on 31 May 2012
reach the silver surface.
The advantages of the Ag-PEG system are numerous and are
particularly interesting for SERS in biologically relevant envi-
ronment. First of all, the PEGylated nanoparticles should be
more stable at the high ionic strength of buffers typically used for
biochemical and biological analysis. The Ag-PEG particles will
not aggregate during incubation with cells. Indeed, these parti-
cles might be internalised by live cells, as it was shown for PEG-
coated SPION (superparamagnetic iron oxide nanoparticles) of
similar dimensions.25 Moreover, the PEG coating can serve as
a linker allowing the particle surface to be enriched with ligands
recognisable by cells for biologically-specific targeting.4
On the other hand, these particles would probably be pro-
tected from aggregation inside cells. Therefore, they would lead
to a more homogeneous intracellular distribution than classical
silver nanoparticles, and their staining would be less disturbing
for cells. This advantage is crucial in view of performing high-
resolution SERS spectral mapping of subcellular compart-
ments.11
The main drawback of the Ag-PEG system is that it would be
limited to the detection of small and mobile analytes able to
diffuse through the polymer coating. From this point of view, it
would probably be impossible to study directly the analyte–
macromolecule complex. On the other hand, one can expect that
the Ag-PEG will be useful for the analysis of ions, drugs and
metabolites in a complex biologically-relevant environment of
variable pH and ionic strength.
Conclusions
The Ag-PEG system developed in this study has a potential for
bioanalytical applications of SERS spectroscopy, in particular
for the SERS spectral imaging of live cells. The intracellular
1872 | Analyst, 2009, 134, 1868–1872
View Online
distribution of PEGylated NPs in cancer cells will be studied
soon.
Additional investigations on the Ag-PEG NPs obtained
through the covalent binding of PEG chains with different
molecular weights and with different functional groups are
underway.
Acknowledgements
The authors acknowledges Region Centre for a postdoctoral
fellowship for A. S. and for the financial support of the research
on functionalized nanoparticles (project ‘NANOMAG’).
References
1 D. Graham and R. Goodacre, Chem. Soc. Rev., 2008, 37, 883.
2 M. J. Banholzer, J. E. Millstone, L. Qin and C. A. Mirkin, Chem. Soc.
Rev., 2008, 37, 885.
3 S. Lal, N. K. Grady, J. Kundu, C. S. Levin, J. B. Lassiter and
N. J. Halas, Chem. Soc. Rev., 2008, 37, 898.
4 X.-M. Qian and S. M. Nie, Chem. Soc. Rev., 2008, 37, 912.
5 W. E. Smith, Chem. Soc. Rev., 2008, 37, 955.
6 N. P. W. Pieczonka and R. F. Aroca, Chem. Soc. Rev., 2008, 37, 946.
7 P. C. Lee and D. P. Meisel, J. Phys. Chem., 1982, 86, 3391.
8 C. E. Talley, J. B. Jackson, C. Oubre, N. K. Grady, C. W. Hollars,
S. M. Lane, T. R. Huser, P. Nordlander and N. J. Halas, Nano
Lett., 2005, 5, 1569.
9 P. Hildebrandt and M. Stockburger, J. Phys. Chem., 1984, 88, 5935.
10 A. Otto, A. Bruckbauer and Y. X. Chen, J. Mol. Struct., 2003, 661,
501.
11 I. Chourpa, F. H. Lei, P. Dubois, M. Manfait and
G. D. Sockalingum, Chem. Soc. Rev., 2008, 37, 993.
12 K. Herve, L. Douziech-Eyrolles, E. Munnier, S. Cohen-Jonathan,
M. Souce, H. Marchais, P. Limelette, P. Warmont, M. L. Saboungi,
P. Dubois and I. Chourpa, Nanotechnology, 2008, 19(46), 465608.
13 W. Yana, R. Wanga, Z. Xua, J. Xua, L. Lina, Z. Shena and Y. Zhoua,
J. Mol. Catal. A: Chem., 2006, 255, 81.
14 C. Luo, Y. Zhang, X. Zeng, Y. Zeng and Y. Wang, J. Colloid Interface
Sci., 2005, 288, 444.
15 M. Popa, T. Pradell, D. Crespo and J. M. Calder on-Moreno, Colloids
Surf., 2007, 303, 184.
16 Z. Chena and L. Gao, Mater. Res. Bull., 2007, 42, 1657.
17 X. Qian, X.-H. Peng, D. O. Ansari, Q. Yin-Goen, G. Z. Chen,
D. M. Shin, L. Yang, A. N. Young, D. Wang and S. Nie, Nat.
Biotechnol., 2008, 26, 83.
18 T. P. Ang and W. S. Chin, J. Phys. Chem. B, 2005, 109, 22228.
19 L. Bo, W. Yang, M. Chen, J. Gao and Q. Xue, Chem. Biodiversity,
2009, 6, 111.
20 H. S. Shin, H. J. Yang, S. B. Kim and M. S. Lee, J. Colloid Interface
Sci., 2004, 274, 89.
21 J. C. Heckel, F. F. Farhan and G. Chumanov, Colloid Polym. Sci.,
2008, 286, 1545.
22 A. Feofanov, S. Sharonov, F. Fleury, I. Kudelina and I. Nabiev,
Biophys. J., 1997, 73, 3328.
23 I. Nabiev, A. Baranov, I. Chourpa, A. Beljebbar, G. D. Sockalingum
and M. Manfait, J. Phys. Chem., 1995, 99, 1608.
24 C. McLaughlin, D. MacMillan, C. McCardle and W. E. Smith, Anal.
Chem., 2002, 74, 3160.
25 T.-J. Chen, T.-H. Cheng, Y.-C. Hung, K.-T. Lin, G.-C. Liu and
Y. M. Wang, J. Biomed. Mater. Res., Part A, 2007, 87, 165.
This journal is ª The Royal Society of Chemistry 2009