Highly Ordered Assemblies of Au

Nanoparticles Organized on DNA

Hidenobu Nakao,

Hiroshi Shiigi,

Yojiro Yamamoto,

Shiho Tokonami,

Tsutomu Nagaoka,

Shigeru Sugiyama,

and Toshio Ohtani*

,
National Food Research Institute, 2-1-12 Kannondai, Tsukuba,
Ibaraki 305-8642, Japan, Department of Applied Chemistry, Faculty of Engineering,
Yamaguchi UniVersity, 2-16-1 Tokiwadai, Ube 755-8611, Japan, and
Research Institute of AdVanced Science & Technology,
Osaka Prefecture UniVersity, 1-2 Gakuen, Sakai 599-8570, Japan
Received August 7, 2003; Revised Manuscript Received August 29, 2003
ABSTRACT
We developed a simple method for highly ordered assemblies of gold nanoparticles (AuNPs) along DNA molecules on substrates, and achieved
assemblies with well-aligned and long-range order by using well-stretched DNA templates. In addition, oxidized aniline-capped AuNPs (AN
AuNPs) prepared in this study were strongly attached to DNA. Two different assembly methods were carried out, and consequently continuous
depositions and necklace-like depositions of ANAuNPs along DNA molecules were achieved.
Although the realization of nanoscale electronics is a subject
of great priority for industrial technology, modern techniques
based on optical lithography have grown increasingly
complicated with attempts to achieve further miniaturization.
Successful routes toward nanoscale electronics include the
use of macromolecules and small molecules as building
blocks for electronic devices. Nanoparticles (NPs), such as
metal and semiconductor colloids with diameters of one to
a few nanometers, are particularly attractive candidates.
1-7
Metal and semiconductor NPs have already been used as
the functional components of nanoscale electronic devices.
4-7
Thus, the ability to organize, interconnect and address NPs
on surfaces is an important hurdle to overcome in order to
fully realize nanoscale electronics. Many natural materials
that have specific binding properties in molecular recognition
and self-assembly can be used as a template to organize and
interconnect assemblies of NPs on surfaces.
8-11
DNA in
particular has been widely investigated as a template because
of its structurally controlled nanowire with 2 nm, well-
defined polymeric sequence and many functionalities, that
can make possible linear assemblies of NPs or other
nanomaterials.
12-19
Although some methods for assembling
NPs onto DNA have been reported, to date those with well-
aligned and long-range order are almost nonexistent.
In this study, we demonstrated a simple method for highly
ordered assemblies of gold NPs (AuNPs) along DNA
molecules on substrates. To securely attach AuNPs to DNA
molecules, preparations of various surface-functionalized
AuNPs have been reported. In numerous other studies, such
AuNPs were prepared by thiolate ligand (cationic thiols
16,17,20
or intercalators
21
) exchange. However, we attempted a one-
step preparation of surface-functionalized AuNPs without
ligand exchange. Novel surface-functionalized AuNPs or
AN-AuNPs were prepared based on the conventional
reduction
22
of HAuCl
4
using aniline as a reducer, so that
AN-AuNPs would have a positive charge and aromatic ring
on the surface (due to the formation of oxidized aniline
during preparation). The simple preparative procedure was
as follows. A 0.1 M aniline (0.5 mL, Wako Pure Chemical
Industry) reducer was added to a 0.03% chloroauric aqueous
solution acid (25 mL, Wako Pure Chemical Industry) then
stirred at 80 C for 20 min. The resulting solution was stored
in polypropylene containers. One milliliter of the stocked
solution was centrifuged at 15 000 rpm for 30 min at 5 C,
then the supernatant was removed. The precipitate was
redispersed in 1 mL of distilled water. The result was AN-
AuNPs (3.0 10
-4
g/L). The UV spectrum of AN-AuNPs
possessed a plasmon resonance peak at 554 nm (Figure 1a).
The maximum of the peak position was shifted 34 nm toward
the bathochromic side, compared with that (about 520 nm)
of generally reported AuNPs coated by surfactants
23
such
as tetra-n-octylammmonium bromide or methyl-tri-n-octyl-
ammonium chloride. The bathochromic shift indicated that
* Corresponding author: phone:+81-29-838-8054, fax: +81-29-838-
7181, e-mail: ohtani@affrc.go.jp.

National Food Research Institute.

Yamaguchi University.

Osaka Prefecture University.

NANO
LETTERS
2003
Vol. 3, No. 10
1391-1394
10.1021/nl034620k CCC: $25.00 2003 American Chemical Society
Published on Web 09/20/2003
AuNPs were coated with a monolayer having a larger
dielectric constant.
23,24
In addition, characterization of pre-
pared particles by electrophoresis analysis and zeta-potential
measurements revealed the presence of positive charges on
surfaces of those. Based on these results, we suggest that
AN-AuNPs have a monolayer of oxidized aniline on
surfaces.
We then experimented with assemblies of AN-AuNPs
organized on DNA molecules. As shown in Figure 2, two
different procedures were used. In Method I, first, DNA-
stretching and fixation were carried out on the surface
according to previously reported methods,
25,26
which resulted
in highly aligned DNA patterns formed on surfaces. Briefly,
a 5 L droplet of a -DNA solution (4.5 ng/L, Wako
Nippon Gene) was deposited on a polycarbonate-coated
coverslip, then sucked up with a pipet. Air-water interface
motion was induced by sucking, and DNA molecules were
stretched and aligned along the central direction of the
droplet. Next, DNA molecules were treated with 10 L AN-
AuNPs solution (3.0 10
-4
g/L) for 5 min, then rinsed in
water. Before treatment with AN-AuNPs, the height (di-
ameter) of DNA molecules imaged by atomic force micros-
copy (AFM) on the surface was 1.0 nm, which well agreed
with that of a single double-stranded DNA in previous
study.
25
Figure 3 shows AFM images of DNA molecules
after treatment of AN-AuNPs. AFM observation revealed
that many DNA molecules on surfaces had contiguous
particles with raised height (Figure 3b), indicating that
observed heights of DNA molecules were 2.14 ( 0.35 nm
(Figure 3c). The majority of the particles that can be
distinguished from the background are 1.56 ( 0.21 nm
(Figure 1b). Consequently, it is considered reasonable and
proper that the increased DNA molecule heights after
treatment were caused by particle deposition. Additionally,
the above result indicates that AN-AuNPs have strong
interaction with DNA molecules. The strong interaction in
this system is considered to be due to the electrostatic
interaction between the negatively charged phosphate back-
bone of DNA and AN-AuNPs with an oxidized aniline shell.
Conceivably, other interactions such as hydrogen-bonding
interactions also might happen between AN-AuNPs and
DNA molecules.
13
It is reasonable to expect that AN-AuNPs
having amino groups interact with nitrogen or oxygen atoms
of the bases that may be exposed in the major or minor
grooves of the double helix. To see whether AN-AuNPs
have specific interaction with DNA, more precise determi-
nation of the chemical nature and structure of the aniline
coating on the AN-AuNPs surfaces is in progress. Figure
3b suggests that AN-AuNPs are uniformly deposited on
almost all DNA molecules. Since the tip radius limits the
resolution of the AFM instrument, the actual deposition might
be less homogeneous. All of the linear structures observed
by AFM also suggested that assemblies of AN-AuNPs on
surfaces were highly oriented over long distances (Figure
3a). Although further optimizations of treatment time and
concentration of AN-AuNPs are necessary to minimize
nonspecific depositions on surfaces, our method makes it
possible to easily achieve highly oriented assemblies of AN-
AuNPs on aligned DNA molecules as templates.
Figure 1. (a) UV spectrum of AN-AuNPs. (b) Histogram showing
the size distribution of AN-AuNPs measured by AFM observa-
tions.
Figure 2. Procedure of assemblies of gold nanoparticles onto DNA.
Figure 3. AFM images of highly ordered assemblies of AN-
AuNPs onto DNA molecules using method I. (a) Image in a large
scan range. (b) Image in a smaller scan range. (c) Scan profile along
the white line of the image b. The height scale is 5 nm in both
images.
1392 Nano Lett., Vol. 3, No. 10, 2003
As shown in Figure 2, we also demonstrated direct
assemblies of AN-AuNPs on surfaces using Method II.
First, we prepared a mixture of 1 L of AN-AuNPs solution
(3.0 10
-4
g/L) and 5 L -DNA solution (4.5 ng/L) and
incubated for 30 min. Next, samples were stretched and fixed
on surfaces according to the above method. AFM images
showed that DNA molecules were well stretched and aligned,
even when combined with AN-AuNPs (Figure 4). It is
interesting to note that AN-AuNPs with larger interparticle
spacing are assembled along DNA molecules like a necklace.
Since DNA molecules (to which AN-AuNPs were already
attached) were stretched significantly by surface tension,
larger interparticle spacing occurred. Interparticle spacing is
obviously larger (Figure 4b) than in Method I (Figure 3b).
Although the actual interparticle spacing is unknown because
resolution is limited by the tip radius of AFM, it seems that
interparticle spacing is nearly uniform along the DNA
molecules. This indicates that DNA stretching induced by
the surface tension is almost constant over the entire length
of DNA. By controlling the surface tension (the speed of
the air-water interface movement or surface hydrophobic-
ity), it might be possible to control the interparticle spacing
of NPs on DNA molecules. Though the bare DNA on
interparticle is invisible after preparation by Method II, it
might be due to the increase in background roughness caused
by sample deposition. From a section profile along the
aligned particles line (Figure 4c), many particle heights (2.97
( 0.35 nm) assembled by using Method II exceeded those
(about 2.0 nm) in the case of Method I. In addition, we could
not observe the presence of particles exceeding about 3 nm
in the background as shown in Figure 4b. From these results,
the presence of the grain in the case of Method II cannot be
regarded as a single particle. Thus, it seems that particles
exceeding about 3.0 nm in height contain more than one
AN-AuNPs. In Method I, since DNA molecules were
already stretched and fixed on surfaces, DNA attaching of
AN-AuNPs is limited to almost one side. On the other hand,
DNA attaching of AN-AuNPs in the solution phase (Method
II) can occur from any direction. Consequently, we consider
that the larger height measured in Method II is caused by
binding of more than one particle surrounding DNA at close
positions.
In summary, we developed simple methods for highly
ordered assemblies of AuNPs, which enabled us to easily
organize AuNPs with well-aligned and long-range order on
DNA molecules. AN-AuNPs prepared in this study strongly
interacted with DNA molecules. Two approaches used in
this study enabled different formations of linear arrays of
AN-AuNPs. Specifically, linear arrays of AN-AuNPs with
interparticle spacing could be organized onto DNA molecules
like a necklace. By depositing different metals or NPs on
interparticle spacing, it will be possible to tune electrical or
optical properties of linear arrays. This method will also be
helpful for investigating and constructing metal nanoparticle
plasmon waveguides,
27
indicating the energy transport
along a nanoparticle chain with a short distance. Assembling
NPs with various properties onto higher-ordered patterns of
DNA molecules could help the further realization of nano-
scale electronics.
Acknowledgment. One of the authors, H.N., is thankful
for Research Fellowships of the Japan Society for the
Promotion of Science for Young Scientists (JSPS).
References
(1) Mirkin, C. A.; Letsinger, R. L.; Mucic, R. C.; Storhoff, J. J. Nature
1996, 382, 607.
(2) Shiigi, H.; Yamamoto, Y.; Yakabe, H.; Tokonami, S.; Nagaoka, T.
Chem. Commun. 2003, 1038.
(3) Torimoto, T.; Yamashita, M.; Kuwabata, S.; Sakata, T.; Mori, H.;
Yoneyama, H. J. Phys. Chem. B 1999, 103, 7799.
(4) Kim, S.-H.; Markovich, G.; Rezvani, S.; Choi, S. H.; Wang, K. L.;
Heath, J. R. Appl. Phys. Lett. 1999, 74, 317.
(5) Berven, C. A.; Clarke, L.; Mooster, J. L.; Wybourne, M. N.;
Hutchison, J. E. AdV. Mater. 2001, 13, 109.
(6) Sato, T.; Ahmed, H.; Brown, D.; Johnson, B. F. J. Appl. Phys. 1997,
82, 696.
(7) Thelander, C.; Magnusson, M. H.; Deppert, K.; Samuelson, L.;
Poulsen, P. R.; Nygrd, J.; Borggreen J. Appl. Phys. Lett. 2001, 79,
2106.
(8) Yamashita, I. Thin Solid Films 2001, 393, 12.
(9) Macmillan, R. A.; Paavola, C. D.; Howard, J.; Chan, S. L.; Zaluzec,
N. J.; Trent, J. D. Nature Mater. 2002, 1, 247.
(10) Lee, S. W.; Mao, C.; Flynn, C. E.; Belcher, A. M. Science 2002,
296, 892.
(11) Scheibel, T.; Parthasarathy, R.; Sawicki, G.; Lin, X.-M.; Jaeger, H.;
Lindquist, S. L. Proc. Natl. Acad. Sci. U.S.A. 2003, 100, 4527.
(12) Coffer, J. L.; Bigham, S. R.; Li, X.; Pinizzotto, R. F.; Rho, Y. G.;
Pirtle, R. M.; Pirtle, I. L. Appl. Phys. Lett. 1996, 69, 3851.
(13) Harnack, O.; Ford, W. E.; Yasuda, A.; Wessels, J. M. Nano Lett.
2002, 2, 919.
(14) Keren, K.; Krueger, M.; Gilad, R.; Ben-Yoseph, G.; Silvan, U.; Braun,
E. Science 2002, 72, 297.
(15) Mertig, M.; Ciacchi, L. C.; Seidel, R.; Pompe, W.; Vita, A. D. Nano
Lett. 2002, 2, 841.
(16) Warnerd, M. G.; Hutchison, J. E. Nature Mater. 2003, 2, 272.
(17) Yonezawa, T.; Onoue, S.; Kimizuka, S. Chem. Lett. 2002, 1172.
(18) Monson, C. F.; Woolley, A. T. Nano Lett. 2003, 3, 359.
(19) Xin, H.; Wooly, A. T. J. Am. Chem. Soc. 2003, 125, 8710.
Figure 4. AFM images of highly ordered assemblies of AN-
AuNPs onto DNA molecules using method II. (a) Image in a large
scan range. (b) Image in a smaller scan range. (c) Scan profile along
particles line from I to II in the image b. Red marks is showing the
particle height indicated by the white arrow in the image b. The
height scale is 5 nm in both images.
Nano Lett., Vol. 3, No. 10, 2003 1393
(20) McIntosh, C. M.; Esposito, E. A., III; Boal, A. K.; Simard, J. M.;
Martin, C. T.; Rottelo, V. M. J. Am. Chem. Soc. 2001, 123, 7626.
(21) Wang, G.; Zhang, J.; Murray, R. W. Anal. Chem. 2002, 74, 4320.
(22) Turkevich, J.; Garton, G.; Sevenson, P. C. J. Colloid Sci. 1954, 9,
26.
(23) Harada, G.; Sakurai, H.; Matsushita, M. M.; Izuoka, A.; Sugawara,
T. Chem. Lett. 2002, 1030.
(24) Okamoto, T.; Yamaguchi, I.; Kobayashi, T. Opt. Lett. 2000, 25, 372.
(25) Nakao, H.; Hayashi, H.; Yoshino, T.; Sugiyama, S.; Otobe, K.;
Ohtani, T. Nano Lett. 2002, 2, 475.
(26) Nakao, H.; Gad, M.; Sugiyama, S.; Otobe, K.; Ohtani, T. J. Am.
Chem. Soc. 2003, 125, 7162.
(27) Maier, S. A.; Kik, P. G.; Atwater, H. A.; Meltzer, S.; Harel, E.; Koel,
B.; Requicha, A. G. Nature Mater. 2003, 2, 229.
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