306 Chem. Mater.

2000, 12, 306-313

Seeding of Colloidal Au Nanoparticle Solutions. 2.

Improved Control of Particle Size and Shape
Kenneth R. Brown, Daniel G. Walter, and Michael J. Natan*
Department of Chemistry, Pennsylvania State University,
University Park, Pennsylvania 16802
Received February 3, 1998. Revised Manuscript Received October 13, 1999

New fields of research in chemistry and physics require improved synthetic techniques
for colloidal metal particles. This paper reports a generally applicable technique for
synthesizing colloidal Au particles of mean diameters between 20 and 100 nm that exhibit
improved monodispersity relative to previously published methods. In this approach (called
“seeding”) Au3+ is reduced on the surface of preformed 12-nm-diameter Au nanoparticles by
introduction of boiling sodium citrate, producing particles highly uniform in size and shape.
A similar procedure, utilizing the reductant NH2OH at room temperature, produces two
populations of particles; the larger population is even more spherical than citrate-reduced
particles of similar size, while the smaller population is very distinctly rod-shaped.

Introduction of these particles in solution,1-5 nanoparticles pre-

pared from metal ions in solution comprise the preferred
The use of nanosized colloidal Au nanoparticles has starting materials.
expanded greatly in recent years. Whereas 10-15 years Many different preparations have been reported for
ago, the predominant use of colloidal Au was in biologi- synthesis of colloidal Au,1,8 including even some that
cal transmission electron microscopy, a wide variety of begin with bulk metal.9 However, most begin with Au3+,
recent papers now describe interesting physical proper- and through use of different reductants, generate
ties1 and possible applications2 that extend far beyond particles with a range of particle sizes. For example,
imaging. For example, Mirkin’s group has developed reducing agents such as NaBH4 or white phosphorus
colorimetric DNA sensors based on colloidal Au.3 More- produce small Au particles (diameter < 10 nm) while
over, organized two-dimensional (2-D) and three- reductants such as ascorbic acid yield colloidal Au
dimensional (3-D) arrays of colloidal Au nanoparticles nanoparticles with diameters larger than 10 nm.10 The
are now occupying the attention of several groups.4
Unless one studies metal nanoparticles one at a time, (4) (a) Andres, R. P.; Bielefeld, J. D.; Henderson, J. I.; Janes, D.
as has recently been described,5 understanding the B.; Kolagunta, V. R.; Kubiak, C. P.; Mahoney, W. J.; Osifchin, R. G.
behavior of solutions and/or surfaces comprising Au Science 1996, 273, 1690-1693. (b) Brust, M.; Walker, M.; Bethell, D.;
Schiffrin, D. J.; Whyman, R. J. Chem. Soc., Chem. Commun. 1994,
nanoparticles is predicated upon having a single size 801-802. (c) Giersig, M.; Mulvaney, P Langmuir 1993, 9, 3408-3413.
(and shape) of particle because, in the nanometer (d) Alivisatos, A. P.; Johnson, K. P.; Peng, X.; Wilson, T. E.; Loweth,
regime, almost every relevant physical property of C. J.; Bruchez, M. P., Jr.; Schultz, P. G. Nature 1996, 382, 609. (e)
Harfenist, S. A.; Wang, Z. L.; Alvarez, M. M.; Vezmar, I.; Whetten, R.
colloidal Au is size-dependent.6 Photolithography7a and L. J. Phys. Chem. 1995, 100, 13904-13910. (f) Mirkin, C. A.; Letsinger,
more recently, nanosphere lithography7b has been used R. L.; Mucic, R. C.; Storhoff, J. J. Nature 1996, 382, 607. (g) Rubin, S.;
Bar, G.; Taylor, T. N.; Cutts, R. W.; Zawodzinski, Jr., T. A. J. Vac.
to prepare surfaces with highly regular metal features. Sci. Technol. A 1996, 14, 1870-1877. (h) Feldstein, M. J.; Keating, C.
However, for materials synthesis and for manipulations D.; Liau, Y.-H.; Natan, M. J.; Sherer, N. F. J. Am. Chem. Soc. 1997,
119, 6638-6647. (i) Grabar, K. C.; Smith, P. C.; Musick, M. D.; Davis,
J. A.; Walter, D. G.; Jackson, M. A.; Guthrie, A. P.; Natan, M. J. J.
* Author whom all correspondence should be addressed via email Am. Chem. Soc. 1996, 118, 1148-1152.
at natan@chem.psu.edu. (5) (a) Emory, S. R.; Nie, S. Science 1997, 275, 1102-1106. (b)
(1) (a) Kreibig, U.; Vollmer, M. Optical Properties of Metal Clusters; Emory, S. R.; Nie, S. Anal. Chem. 1997, 69, 2631-2635. (c) Klein, D.
Springer-Verlag: Berlin, 1995. (b) Schmid, G. Chem. Rev. 1992, 92, L.; McEuen, P. L.; Bowen, J. E.; Roth, R.; Alivisatos, A. P. Appl. Phys.
1709-1727. (c) Mulvaney, P. Langmuir 1996, 12, 788-800. (d) Kume, Lett. 1996, 68, 2574-2576.
T.; Nakagawa, N.; Hayashi, S.; Yamamoto, K. Solid State Commun. (6) (a) Freund, P. L.; Spiro, M. J. Phys. Chem. 1985, 89, 1074-
1995, 93, 171-175. (e) Farbman, I.; Levi, O.; Efrima, S. J. Chem. Phys. 1077. (b) Zeman, E. J.; Schatz, G. C. J. Phys. Chem. 1987, 91, 634-
1992, 96, 6477-6485. (f) Chernov, S. F.; Zakharov, V. N. J. Mod. Opt. 643. (c) Quinten, M.; Kreibig, U. Surf. Sci. 1986, 172, 557-577.
1989, 36, 1541-1544. (g) Hache, F.; Ricard, D.; Flytzanis, C.; Kreibig, (7) (a) Liao, P. F.; Bergman, J. G.; Chemla, D. S.; Wokaun, A.;
U. Appl. Phys. A 1988, 47, 347-357. Melngailis, J.; Hawyrluk, A. M.; Economou, N. P. Chem. Phys. Lett.
(2) (a) Fukumi, K.; Chayahara, A.; Kadono, K.; Sakaguchi, T.; 1981, 82, 355-359. (b) Hulteen J. C.; Van Duyne, R. P. J. Vac. Sci.
Horino, Y.; Miya, M.; Fujii, K.; Hayakawa, J.; Satou, M. J. Appl. Phys. Technol., A 1995, 13, 1553-8.
1994, 75, 3075-3080. (b) Lipscomb, L. A.; Nie, S.; Feng, S.; Yu, N.-T. (8) (a) Torigoe, K.; Esumi, K. Langmuir 1992, 8, 59-64. (b) Esumi,
Chem. Phys. Lett. 1990, 170, 457-461. (c) Angel, S. M.; Katz, L. F.; K.; Sato, N.; Torigoe, K.; Meguro, K. J. Colloid Interface Sci. 1992,
Archibald, D. D.; Honigs, D. E. Appl. Spectrosc. 1989, 43, 367-372. 149, 295-298. (c) Leff, D. V.; Ohara, P. C.; Heath, J. R.; Gelbart, W.
(d) Kneipp, K.; Dasari, R. R.; Wang, Y. Appl. Spectrosc. 1994, 48, 951- M. J. Phys. Chem. 1995, 99, 7036-7041. (d) Duff, D. G.; Baiker, A.
955. Langmuir 1993, 9, 2301-2309. (e) Duff, D. G.; Baiker, A. Langmuir
(3) (a) Elghanian, R.; Storhoff, J. J.; Mucic, R. C.; Letsinger, R. L.; 1993, 9, 2310-2317.
Mirkin, C. A. Science 1997, 277, 1078-1081. (b) Storhoff, J. L.; (9) (a) Beans, H. T.; Eastlack, H. E. J. Am. Chem. Soc. 1915, 37,
Elghanian, R.; Mucic, R. C.; Mirkin, C. A.; Letsinger, R. L. J. Am. 2667-2683. (b) Nedderson, J.; Chumanov, G.; Cotton, T. M. Appl.
Chem. Soc. 1998, 120, 1959-1964. Spectrosc. 1993, 47, 1959-1964.

10.1021/cm980065p CCC: $19.00 © 2000 American Chemical Society

Published on Web 12/29/1999
Seeding of Colloidal Au Nanoparticle Solutions Chem. Mater., Vol. 12, No. 2, 2000 307

most widely studied reductant is sodium citrate;1,11 by to 10 mL. A total of 0.1 mL of this solution was mixed with
varying the citrate:Au ratio, it is possible to prepare 0.1 mL of 2% HNO3 and 0.8 mL of H2O for analysis. Standards
colloidal particles with diameters (d) ranging from 10 were prepared from a 100 ppm stock solution of Au ion (Perkin-
Elmer), from which 0.1 mL was mixed with 0.1 mL of 30 µL
to 150 nm.12 Unfortunately, for d > 30 nm, the mono- of HNO3/30 µL of HCl and diluted to 10 mL. Different volumes
dispersity becomes poor and the ellipticity (G)sthe ratio of 1 ppm stock were mixed with 0.1 mL of 2% HNO3 and
of the major to minor axisssignificantly exceeds unity, diluted to 1.0 mL to yield standards between 25 and 200 ppb.
the value for a sphere. As a result, these particles are AA samples were burned at 130 °C for 60 s, 1000 °C for 45 s,
of limited value for nanometer-scale architecture. In- and atomized at 1800 °C with a 5-s reading window. No salt
deed, of the many published routes to colloidal Au, none interference was observed.
produce large (>50 nm) particles with good monodis- The 2.6-nm-Diameter Colloidal Au. One milliliter of 1%
HAuCl4 was added to 90 mL of H2O at room temperature (20-
persity. Their utility is further restricted by the low
23 °C). After 1 min of stirring, 2.00 mL of 38.8 mM sodium
particle concentrations generated by these methods. citrate was added. One minute later, 1.00 mL of fresh 0.075%
We describe herein the synthesis and characterization NaBH4 in 38.8 mM sodium citrate was added. The colloidal
of large colloidal Au particles prepared by the “seeding” solution was stirred for an additional 5 min and stored in a
approach, an adaptation of a method described in the dark bottle at 4 °C.
patent literature.13 In the reported method, 2.6-nm- Citrate-Seeded Colloidal Au. To a stirred solution of boiling
diameter colloidal Au nanoparticles (“seeds”) prepared 0.01% HAuCl4, seed colloids (either 2.6-nm-diameter or 12-
nm-diameter) were added coincidentally with the addition of
by BH4- reduction are grown to larger sizes by addition
38.8 mM sodium citrate (final concentration ) 0.17 mM). This
of a boiling solution of HAuCl4 and citrate. This ap- mixture was boiled for 15 min and stirred for an additional
proach produces large colloidal Au particles more pre- 10 min while cooling. Citrate-seeded colloids were prepared
dictably and reproducibly than citrate reduction, but in volumes ranging from 50 mL to 500 mL, with different
because the seeds are highly polydisperse (40% standard diameters were generated by changing the volume of seed
deviation), the monodispersity is comparable. We rea- colloid added.
soned that use of more monodisperse seeds (e.g., 12 ( NH2OH-Seeded Colloidal Au. These syntheses utilized stock
1.5 nm diameter) should yield larger particles particles solutions of 1% HAuCl4 and 40 mM H2NOH, diluted to final
concentrations as described in the figure captions. Except
with narrower size distributions, a notion confirmed by where noted, seed particles were added to stirred mixtures of
experiment. We also describe a superior new synthetic NH2OH and HAuCl4.
protocol, based on room-temperature seeding of colloidal Instrumentation. Optical spectra were obtained on an
Au by NH2OH/Au3+.14,15 This method is consistent and HP8452A diode array UV-vis spectrophotometer with a
reproducible enough to predict, a priori, final colloidal deuterium lamp (350-820-nm range, 2-nm resolution). Graph-
diameters within a few nanometers. Iterative seeding ite furnace atomic absorption was performed with a Perkin-
using this approach can be used to rapidly produce large Elmer 1100B spectrophotometer, an HGA 700 power supply,
an AS-70 autosampler and a Au lamp (Perkin-Elmer). Equip-
colloidal Au nanoparticle solutions that exhibit excellent ment and protocols for acquisition and analysis of transmission
monodispersity. Interestingly, repetitive seeding with electron microscopy (TEM) images have been described else-
NH2OH leads to formation of a small percentage of where.18,19 Software protocols that allowed 2-D clusters of
cylindrical, high aspect ratio rods. Note that such rods particles to be separated into individual entities (or when this
have been generated electrochemically in the pores of was not possible, eliminated from consideration) were em-
membranes.16,17 ployed.

Experimental Section Results and Discussion

Materials. HAuCl4, Na3citrate, NaBH4, HCl, HNO3, H2SO4, Citrate Seeding. As part of a study aimed at
and aqueous H2O2 were obtained from Sigma-Aldrich, Fisher preparation of protein:Au colloid complexes, a thorough
(Acros), or VWR and used without further purification. H2O study of the physical properties Au nanoparticles made
was >18 MΩ from a Barnstead Nanopure water purification by the Frens method (i.e., direct reduction of Au3+ by
system.
citrate12) has been published.20 By varying the citrate:
Methods. General protocols for synthesis, characterization,
handling, and storage of colloidal Au solutionssincluding for
Au3+ ratio, Au nanoparticles with diameters from 10 to
17 nM, 12-nm-diameter particlesshave been previously de- 70 nm were prepared and characterized by TEM and
scribed.18 Samples for atomic absorption (AA) were prepared UV-vis spectrophotometry. Four parameters were used
by digesting 0.1 mL of colloidal Au solution with 30 µL of to describe each preparation of colloid: the wavelength
concentrated HNO3 and 30 µL of concentrated HCl, and diluted
(18) (a) Grabar, K. C.; Freeman, R. G.; Hommer, M. B.; Natan, M.
(10) Slot, J. W.; Geuze, H. J. J. Cell Biol. 1981, 90, 533-536. J. Anal. Chem. 1995, 67, 735-743. (b) Bright, R. M.; Walter, D. G.;
(11) Turkevich, J.; Hillier, J.; Stevenson, P. C. Discuss. Faraday Musick, M. D.; Jackson, M. A.; Allison, K. J.; Natan, M. J. Langmuir
Soc. 1951, 11, 55. 1996, 12, 810-817. (c) Freeman, R. G.; Grabar, K. C.; Allison, K. J.;
(12) (a) Frens, G. Nature 1972, 241, 20-22. (b) Frens, G. Kolloid- Bright, R. M.; Davis, J. A.; Guthrie, A. P.; Hommer, M. B.; Jackson,
Z. u. Z. Polymere 1972, 250, 736-741. M. A.; Smith, P. C.; Walter, D. G.; Natan, M. J. Science 1995, 267,
(13) Schutt, E. G. Eur. Patent Application 90317671.4, filed Sep- 1629-1632. (d) Grabar, K. C.; Allison, K. J.; Baker, B. E.; Bright, R.
tember 25, 1990. M.; Brown, K. R.; Freeman, R. G.; Fox, A. P.; Keating, C. D.; Musick,
(14) Stremsdoerfer, G.; Perrot, H.; Martin, J. R.; Clechet, P. J. M. D.; Natan, M. J. Langmuir 1996, 12, 2353-2361.
Electrochem. Soc. 1988, 135, 2881-2885. (19) Grabar, K. C.; Brown, K. R.; Keating, C. D.; Stranick, S. J.;
(15) Brown, K. R.; Natan, M. J. Langmuir 1998, in press. Tang, S.-L.; Natan, M. J. Anal. Chem. 1997, 69, 417-477.
(16) Martin, C. R. Science 1994, 266, 1961-1966. (b) Foss, C. A.; (20) (a) Goodman, S. L.; Hodges, G. M.; Trejdosiewicz, L. K.;
Tierney, M. J.; Martin, C. R. J. Phys. Chem. 1992, 96, 9001-9007. (c) Livingston, D. C. J. Microsc. 1981, 123, 201-213. (b) Faulk, W. P.;
Hornyak, G. L.; Patrissi, C. J.; Martin, C. R. J. Phys. Chem. 1997, Taylor, G. M. Immunochemistry 1971, 8, 1081-1083. (c) Horisberger,
101, 1548-1555. (d) Foss, C. A.; Nornyak, G. L.; Stockert, J. A.; Martin, M. Biol. Cell. 1979, 36, 253-258. (d) Hayat, M. A., Ed.; Colloidal
C. R. J. Chem. Phys. 1994, 98, 2963-2971. Gold: Principles, Methods, and Applications; Academic Press: San
(17) (a) Al-Mawlawi, D.; Liu, C. Z.; Moskovits, M. J. Mater. Res. Diego, 1989; Vols. 1 and 2. (e) Beesley, J. E. Colloidal Gold: A New
1994, 9, 1014-18. (b) Preston, C. K.; Moskovits, M. J. Phys. Chem. Perspective for Cytochemical Marking; Oxford University: Oxford,
1993, 97, 8495-8503. 1989; Vol. 17.
308 Chem. Mater., Vol. 12, No. 2, 2000 Brown et al.

Table 1. Physical Properties of Colloidal Au Nanoparticles Prepared by Direct Citrate Reduction and by Citrate
Seeding of 2.6-nm-Diameter and 12-nm-Diameter Colloidal Au Solutions
λmax peak width
batch major axis × minor axisa Gb (nm) (nm) source of colloid
small seeds 2.6 (1.0) × 2.0 (0.8) 1.30 514 104 this work
large seeds 12.6 (1.1) × 11.5 (1.0) 1.10 518 84 this work
Synthetic Method
direct citrate 1 21 (2.3) × 19 (2.3)c 1.13 524 83 ref 20
2 22.3 (5.0) × 18.6 (2.4) 1.20 528 116 this work
3 25 (3.3) × 21 (3.3) 1.19 524 90 ref 20
4 31.7 (7.6) × 24.5 (3.9) 1.29 530 136 this work
5 35 (4.0) × 26 (4.0) 1.33 530 108 ref 20
6 44 (6.9) × 33 (6.9) 1.34 528 104 ref 20
7 44.9 (9.5) × 36.4 (5.6) 1.23 524 118 this work
8 48 (10.5) × 37 (10.5) 1.31 535 98 ref 20
9 56 (8.4) × 41 (8.4) 1.37 535 147 ref 20
seeded, large 1 19.3 (1.7) × 16.4 (1.1) 1.18 520 84 this work
2 21.4 (2.6) × 18.5 (1.9) 1.16 522 80 this work
3 25.0 (2.2) × 21.3 (1.6) 1.17 524 80 this work
4 28.7 (2.6) × 24.3 (2.0) 1.18 526 76 this work
5 31.1 (3.5) × 26.0 (2.4) 1.20 526 82 this work
6 38.4 (4.7) × 31.8 (2.7) 1.21 528 78 this work
7 44.5 (5.8) × 36.8 (4.1) 1.21 530 84 this work
8 53 (4.8) × 43 (3.1) 1.23 534 92 this work
9 64 (6.3) × 51 (3.8) 1.26 545 112 this work
10 72 (9.4) × 54 (5.2) 1.33 542 152 this work
11 76 (11) × 56 (6.0) 1.36 538 116 this work
12 91 (14) × 68 (8.2) 1.35 550 220 this work
seeded, small 1 35.8 (8.7) × 30.6 (7.3) 1.17 548 188 this work
2 36.4 (5.2) × 29.6 (3.1) 1.23 530 96 this work
3 43.0 (7.8) × 36.8 (6.2) 1.17 534 132 this work
4 53 (5.2) × 44 (3.7) 1.21 542 108 this work
5 56 (7.2) × 45 (4.6) 1.25 544 144 this work
6 61 (10) × 49 (7.5) 1.24 548 164 this work
7 75 (17) × 60 (11) 1.24 548 216 this work
8 93 (20) × 68 (11) 1.37 572 300 this work
9 108 (38) × 77 (20) 1.40 548 300 this work
10 111 (27) × 81 (17) 1.38 616 404 this work
a Values in parentheses are standard deviations; all values are in nanometers. b Ellipticity, as defined in text. c Reference 19 gives a

single standard deviation for a given preparation of particles, so it is used for both axes.

of maximum absorbance (λmax) and peak width at half 40 and 30 nm indicates the tops of the particles are 30
max (PWHM) from the UV-vis data, and the mean nm above the surface.21
particle diameter (d) and ellipticity (G) from TEM data. We have compared d, G, λmax, and PWHM for Au
Due to the asymmetry of the colloidal Au surface nanoparticles made by the Frens method to those
plasmon band,2a PWHM is defined straightforwardly as prepared by seeding (Figure 1 and Table 1). In the
twice the difference between λmax and the λ of half- approach described in the patent literature,13 the di-
maximal absorbance to the red of the λmax. mensions of existing Au nanoparticles (the “seeds”) are
In contrast, the mean particle diameter d is an increased by mixing with additional reductant (i.e.,
oversimplification in that no colloidal Au particles are citrate) and Au3+. In practice, a mixture of citrate and
purely spherical: TEM images show the presence of colloidal Au is added to a solution of boiling, dilute
major and minor axes (the ellipticity G is the ratio HAuCl4 in H2O. What makes this method succeed (or
major:minor axial ratio).19 Accordingly, while d is used fail) is the relative rate of new particle formation (via
herein in reference to the dimensions of the major axis, Au3+ reduction) in solution versus the rate of Au3+
all particles are described by mean major and minor reduction on the surface of existing particles. If the
axes (e.g., Table 1). The TEM images only two of the latter is much greater than the former, then particle
three dimensions in a particle, and it is assumed that growth will occur at the expense of new particle
the dimensions of the third are the same as one of the nucleation. From the volumes of work on production of
two imaged. The question arises as to whether the colloidal Au nanoparticles for biological applications,1
particles are prolate (cigar-shaped; third axis ) minor) it is well-appreciated that reduction of Au3+ on any
or oblate (pancake-shaped; third axis ) major). While small particles is rapid, explaining the need for filtered
Goodman et al.20 state that colloidal Au nanoparticles solutions and scrupulously clean glassware for the
are oblate, both logic and experiment suggest that the production of colloidal Au. It is thus reasonable to
reverse is true: particles on TEM grids adsorb so as assume that new particle nucleation can be averted by
maximize their contact with the surface, meaning the seeding. The absence of new particle nucleation is
axis normal to the surface must be minor. Since minor especially critical for growth of large particles, because
and major axes are seen in TEM images, there must be in the Frens method, as particles are growing larger,
two minor axes, making the particles prolate. In accord new (small) particles can be forming.
with this reasoning, atomic force microscopy images of
Au nanoparticles on glass slides known to have axes of (21) Lyon, L. A.; Natan, M. J. Unpublished results.
Seeding of Colloidal Au Nanoparticle Solutions Chem. Mater., Vol. 12, No. 2, 2000 309

particle diameter,23 PWHM can be used as a rapid

diagnostic for the quality of citrate-derived, small
diameter colloidal Au preparations. For example, in our
experience, preparations of 12-nm-diameter colloidal Au
with PWHM >85 nm invariably exhibit roughly 2-fold
higher standard devations in d than those with PWHM
<85 nm (and are thus usually discarded).
Comparison of the physical properties of colloidal Au
prepared by seeding with that made by direct citrate
reduction indicates the superiority of the former (Table
1). In particular, G is significantly greater for colloidal
Au made by direct citrate reduction: several batches
clustered around d ) 40 nm all had G g 1.3, while the
analogous particles made by seeding exhibited G values
≈ 1.2. Even particles in the 20-30 nm diameter range
made by direct citrate reduction were more elliptical
than the corresponding seeded particles. Moreover,
PWHM for seeded particles are consistently lower than
for particles made by direct citrate reduction. For the
three sizes of 12-nm seeded particles shown in Figure
2, the PWHM are nearly identical, and the only signifi-
cant difference is the aforementioned shift in λmax.
Colloidal Au solutions made by direct citrate reduction
exhibit broadened PWHM over the entire range of
particle diameters.
The high degree of ellipticity in colloidal Au made by
Figure 1. Comparison of λmax (top), ellipticity (G) (middle), direct citrate reduction arises from two components:
and peak width at half-max (PWHM) (bottom) versus major differential reduction rates at sites on the particle, and
axis diameter for colloidal Au nanoparticles prepared by direct differential diffusion to parts of the particles. High-
citrate reduction of Au3+ ([) and by citrate-based seeding of resolution TEM studies have invariably shown the
2.6-nm-diameter (O) and 12-nm-diameter (b) colloidal Au. presence of crystalline faces in small colloidal Au
nanoparticles.24 If reduction of Au3+ is substantially
faster (or slower) on certain faces than on others, then
anisotropic particle growth can be expected. In this
regard, Wokaun has shown that under certain condi-
tions, highly anisotropic colloidal Au nanoparticles can
be prepared;25 likewise, production of cubic colloidal
particles has recently been described.26 Another factor
that could favor elliptical particles is the enhanced
amount of Au3+ reaching the ends of the ellipse: flux
to a hemisphere is enhanced relative to flux to a plane.27
While this factor is more important in unstirred solu-
tions, even in stirred solutions it could contribute to
increased growth at the hemispherical “ends” of a
Figure 2. Optical spectra for solutions of 19-, 25-, and 53- prolate spheroidal particle.
nm-diameter Au nanoparticles grown by citrate seeding of 12- Interestingly, the standard deviations of the axial
nm-diameter colloidal Au. Size and shape data for these dimensions are substantially lower for particles seeded
particles are described in Table 1. with 12-nm-diameter colloidal Au than for particles
made by direct citrate reduction or 2.6-nm-diameter
Several general trends are evident by inspection of seeding For example, in batch 9 of the particles made
plots of λmax, ellipticity, and peak width at half- by direct citrate reduction, the major axis is 56 ( 8.4
maximum (PWHM) versus particle diameter for col-
loidal Au made by Frens’ method and by seeding using (22) (a) Creighton, J. A.; Blatchford, C. G.; Albrecht, M. G. J. Chem.
2.6- and 12-nm-diameter seeds (Figure 1). For each type Soc., Faraday Trans. 2 1979, 75, 790-798. (b) Blatchford, C. G.;
of particle, λmax increases with increasing size, although Campbell, J. R.; Creighton, J. A. Surf. Sci. 1982, 120, 435-455.
(23) Bohren, C. F.; Huffman, D. R. Adsorption and Scattering of
not dramatically. Increasing the diameter of a colloidal Light by Small Paricles; John Wiley & Sons: New York, 1983.
Au nanoparticle from 19 to 53 nm, nearly a factor of 3, (24) Duff, D. G.; Curtis, A. C.; Edwards, P. P.; Jefferson, D. A.;
leads only to a 14-nm red shift in λmax (Figure 2); red Johnson, B. F. G.; Kirkland, A. I.; Logan, D. E. Angew. Chem., Int.
Ed. Engl. 1987, 26, 676-678.
shifts of 150 nm or more can be realized by aggregation (25) Weisner, J.; Wokaun, A. Chem. Phys. Lett. 1989, 157, 569-
of small particles.18 However, increasing particle diam- 575.
(26) Ahmadi, T. S.; Wang, Z. L.; Henglein, A.; El-Sayed, M. A. Chem.
eters lead to increasing ellipticities and to significantly Mater. 1996, 8, 1161-1163.
broadened optical spectra. The latter has been shown (27) (A) Bard, A. J.; Faulkner, L. R. Electrochemical Methods:
to result in large part from Rayleigh light scattering Fundamentals and Applications; John Wiley & Sons: New York, 1980.
(b) Amatore, C. In Physical Electrochemistry. Principles, Methods and
from particles with diameters > λ/20.22 It should be Applications; Rubinstein, I., Ed.; Marcel Dekker: New York, 1995; pp
noted that because light scattering is so sensitive to 131-208.
310 Chem. Mater., Vol. 12, No. 2, 2000 Brown et al.

Figure 4. Absorbance at 526 nm measured every second (0.1-s

integration time) after additions of 0.1 mL of 0.1% HAuCl4
(at the times indicated) to an unstirred mixture of 3.2 mL of
Figure 3. Optical spectra (0.1-s integration time) of a mixture
0.01% HAuCl4/0.1 mL of 40 mM H2NOH/0.050 mL of 17 nM,
of 3.2 mL of 0.01% HAuCl4/0.1 mL of 40 mM H2NOH after 20
12-nm-diameter colloidal Au.
min (a), and 1 (b), 6 (c), 11 (d), and 56 s (e) after addition
(without stirring) of 0.050 mL of 17 nM, 12-nm-diameter
colloidal Au.

nm. For small seeds of the corresponding size (batch 5)

d ) 56 ( 7.2 nm, while for batch 8 of the large seeds,
d ) 53 ( 4.8 nm. The lower dispersity in particle size
for 12-nm-seeded particles becomes even more signifi-
cant for larger particles (e.g., large seed batch 9 vs small
seed batch 6, large seed batch 10 vs small seed batch 7,
and so on).
The increased dispersity in particle size for particles
made using 2.6-nm diameter seeds simply reflects the
poor monodispersity and increased ellipticity of the
seeds themselves (Table 1 and Supporting Information).
Despite many attempts, we have found that the 2.6-
nm-diameter colloidal particles always have ∼1-nm
standard deviations in diameter. While this about the
same absolute standard deviation in diameter measured
for good preparations of 12-nm-diameter particles, it is
quite large as a percentage of particle size. Furthermore,
the measured ellipticity is 1.3 (versus 1.1 for 12-nm-
diameter particles). Accordingly, seeding with 2.6-nm Figure 5. Representative TEM image of enlarged colloidal
particles leads to more dispersion in particle size, Au particles prepared by a two-step NH2OH seeding of colloidal
reflected by larger standard deviations in diameter and Au (0.05 mL of 16.8 nM, 12-nm-diameter colloidal Au added
by very substantial increases in peak widths (Table 1). to an unstirred mixture of 2.4 mL of 0.01%HAuCl4/0.1 mL of
40 mM NH2OH, then 0.1 mL of 1% HAuCl4 added after ∼2
In short, citrate seeding 12-nm-diameter colloidal Au min).
nanoparticles yields sols with improved physical proper-
ties relative to direct particle production by citrate adsorbed Au+ ions is possible.28 Similarly, the reductant
reduction of Au3+ or by seeding with 2.6-nm-diameter hydroxylamine (NH2OH) has previously been shown to
seeds. The disparity in quality of colloidal preparations favor reduction of Au ions at metallic Au surfaces at
is especially prevalent for d > 40 nm. room temperature over reduction in solution,14 and we
Hydroxylamine Seeding. Drawbacks of citrate have described in a preliminary fashion the application
seeding include the need for boiling H2O and the need of NH2OH seeding to colloidal Au nanoparticles.15
to keep the Au3+ and the reductant apart; ideally, At room temperature, mixtures of NH2OH and Au3+
seeding could be carried out at room temperature, and (added as HAuCl4) do not lead to formation of colloidal
initiated upon introduction of seeds to premixed solu- Au nanoparticles in solution. Thus, the optical spectrum
tions of Au3+ and reductant (or introduction of Au3+ to of a mixture of 0.01% HAuCl4 and 40 mM H2NOH is
a mixture of reductant and seeds). This can be ac- featureless after 20 min (spectrum a of Figure 3).
complished by a process with a solution nucleation rate Addition of 50 µL of a solution containing 12-nm-
constant of zero at room temperature and a surface diameter colloidal Au seeds leads immediately to sig-
nucleation rate constant significantly greater than zero nificant growth in the intensity of the colloidal Au
at room temperature. In Au electroless metal plating, surface plasmon band (spectra b-e in Figure 3). Changes
these requirements are satisfied by chelation of Au+ to
cyanide, which moves the solution reduction potential (28) Kumar, A.; Biebuyck, H. A.; Whitesides, G. M. Langmuir 1994,
too far negative for formation of Au0, so that only 10, 1498-1511.
Seeding of Colloidal Au Nanoparticle Solutions Chem. Mater., Vol. 12, No. 2, 2000 311

Figure 6. TEM images of colloidal Au grown by iterative NH2OH seeding. Major axis dimensions for spherical particles are as
follows: 13 nm (panel A) [starting material]; 18 nm (panel B); 32 nm (panel C); 41 nm (panel D); 56 nm (panel E); and 116 nm
(panel F). Detailed particle size analysis is given in Table 2; optical spectra and synthetic protocols for each set of particles are
available in the Supporting Information.

in the optical spectra are essentially complete after 11 increase in absorbance,29 followed by a slight decrease
s, since a spectrum recorded after 56 s is essentially and then a leveling off. Each subsequent addition of
identical, as are spectra recorded every 5 s between Au3+ leads to an increase in absorbance, and to a new
these times (data not shown). Since the formation of new (higher) level. Thus, HAuCl4 can be made the limiting
particles is precluded by the control experiment de- reagent in NH2OH-seeded growth of 12-nm-diameter
scribed by spectrum a, the spectral changes suggest colloidal Au: introduction of a known quantity of Au3+
enlargement of the seed particles. If so, cessation in to a solution containing colloidal Au nanoparticles and
particle growth must result from depletion of Au3+. an excess of NH2OH leads to particle growth, until the
This notion is confirmed by the data in Figure 4, Au3+ is consumed.
which plots the absorbance at 526 nm over time as This approach to seeded particle growth is attractive
aliquots of HAuCl4 are added to a prereacted mixture in three respects. First, particle enlargement can be
of 0.01% HAuCl4, NH2OH, and 12-nm-diameter colloidal carried out at room temperature. Second, since Au3+ can
Au. At t ) 0, the absorbance A ≈ 1, indicating that the be made limiting, it should be possible to grow particles
Au3+ already in solution has already been consumed to
enlarge the 12-nm-diameter colloidal Au particles. After (29) The dips in the first seconds after each addition are due to the
addition of Au3+, there is a nearly instantaneous turbulence caused by pipet introduction into the cuvette.
312 Chem. Mater., Vol. 12, No. 2, 2000 Brown et al.

Table 2. Particle Size and Shape Analysis for Iteratively NH2OH-Seeded Colloidal Au Nanoparticles
Figure 6 number mean d median dc G predicted sizec,d
panela sized major × minorb,c major × minor (mean) major × minor
Totale
A 1322 13.0 (1.4) × 11.8 (1.2) 12.8 × 11.7 1.10
B 2829 18.0 (2.7) × 16.3 (2.3) 17.8 × 16.1 1.10
C 2263 35.1 (9.0) × 28.4 (3.6) 32.8 × 28.6 1.24
D 3363 51.1(28.6) × 36.1 (5.4) 41.7 × 37.0 1.42
E 917 65.8(31.0) × 49.5 (7.0) 56.0 × 49.9 1.33
Ff g g g g
Spherese
A 1183 12.9 (1.3) × 11.9 (1.2) 12.7 × 11.7 1.08
B 2630 17.9 (2.5) × 16.4 (2.3) 17.7 × 16.2 1.09 21.0 × 19.0
C 1964 32.4 (3.2) × 29.2 (2.7) 32.3 × 29.0 1.11 33.7 × 30.3
D 2839 41.1 (4.0) × 37.7 (3.5) 41.0 × 37.6 1.09 45.5 × 41
E 799 56.2 (5.4) × 51.0 (5.1) 55.3 × 50.4 1.10 59 × 53
F 1443 116 (11) × 102 (10) 116 × 102 1.14 121 × 103
Rodse
A 120 14.2 (2.1) × 11.6 (1.7) 13.7 × 11.2 1.22
B 100 20.8 (4.3) × 15.1 (1.9) 19.8 × 14.8 1.38
C 200 58.9 (12.8) × 21.4 (3.2) 57.3 × 21.0 2.75
D 399 122 (32.2) × 25.3 (3.9) 127.0 × 24.6 4.82
E 80 145 (42.4) × 35.0 (7.0) 163.5 × 33.0 4.14
F 200 236 (60.3) × 77.1 (9.6) 233.6 × 74.2 3.06
a For convenience, solutions of seeded particles have been named according to their TEM image in Figure 6. b Numbers in parentheses

refer to standard deviations. c In nanometers. d With the exception of B, whose predicted size was based on the actual size of A, predictions
were based on predicted sizes from the previous iteration. e “Total” refers to all the particles in a given batch of colloid, while “spheres”
and “rods” refer those subsets of particles possessing the corresponding shapes. Particles with G > 1.2 were called rods. f F represents a
two-step iteration from E. The predicted dimensions for the hidden iteration (E′) was 84 × 72 nm. g The pronounced bimodality of the
particle size distribution precluded meaningful calculation of mean diameter and ellipticity.

to a prespecified size. Finally, NH2OH/Au3+ seeding is (i.e., with limited Au3+). If, on the other hand, the rods
well-suited for growth of immobilized Au nanoparticles, are formed by highly face-selective surface reactions,
because a colloidal Au-derivatized surface can be easily then stepwise growth will simply lead to stepwise rod
immersed into a solution of HAuCl4/NH2OH.15,30 expansion.
These advantages notwithstanding, the physical prop- Iterative, stepwise seeding should also favorably
erties of NH2OH/Au3+-seeded colloidal Au particles impact the size and shape distribution of the spherical
ultimately determine the merit of this approach. Figure particles. Figure 1 and Table 1 show clearly that single-
5 shows a TEM image of colloidal particles produced step enlargement of 2.6- or 12-nm-diameter colloidal Au
by a NH2OH/Au3+ seeding procedure analogous to that particles diameters below 40 nm is well-behaved; for
described for the data in Figure 3. Two aspects of the larger particles, the PWHM and ellipticity seem to rise
image are striking: the excellent monodispersity of the exponentially. Growing large particles in multiple steps
spherical particles (51 ( 5.2 nm × 46 ( 4.7 nm), and should lead to minimization of these effects.
the presence of a distinct population (5-10%) of colloidal The results of an iterative seeding experiment are
Au rods (141 ( 38 nm × 31 ( 4.6 nm) with ellipticities shown in Figure 6, which depicts TEM images of the
far greater (∼4.5) than any particles derived from citrate starting 12-nm-diameter seed particles and those re-
reduction of Au3+, whether seeded or not. The standard sulting from five iterations, and in Table 2, a detailed
deviation of the major and minor axes of the spherical particle size analysis. (The experimental protocols for
particles appears equivalent to that produced by citrate iterative seeding and the optical spectra of the resulting
seeding with 12-nm-diameter colloidal Au. In contrast, colloidal Au solutions can be found in the Supporting
the length of the rods’ major axes is highly variable, Information.) Perusal of Figure 6, which shows spherical
ranging from 90 nm to >200 nm, while the rods’ minor particles spanning nearly an order of magnitude in size
axes shows the same small standard deviation (4.7 nm) (∼12-13 nm in panel A to over 100 nm in panel F) leads
as the spherical particles. More importantly, the minor to two immediate conclusions. The first is that the
axis is more than three standard deviations shorter than colloidal Au rods are definitely not fused or aggregated
the minor axis of the dominant population, suggesting spheres. This is easily confirmed by examination of
that the rods do not form by fusion of spheres. Figure 7, an enlarged TEM image of panel E, which
The origin of these rods is of great interest, particu- shows that the rods are solid and are significantly
larly in view of theoretical prediction6b and of recent smaller in the minor axis than surrounding quasi-
experimental confirmation5a,b of intense electromagnetic spherical particles. Another interesting point that can
fields at the ends of highly prolate spheroids of Au and be gleaned from Figure 7 is that none of the rods are
Ag. If the process of rod formation has a strong diffu- blunt-ended: both ends are hemispherical, some almost
sional component (as described above for large colloidal perfectly so. The second obvious finding from Figure 6
Au nanoparticles made by the Frens method), it should is that there is no continuum from spherical to rod-
be minimized by growing the particles in small steps shaped particles; rather, there are two quite distinct
populations, with nothing in between. Figure 8, a
(30) Brown, K. R.; Natan, M. J. Chem. Mater. 2000, 12, 314- histogram showing the number of particles (in the
323 (subsequent paper in this journal). colloidal preparation imaged in panel D) with a given
Seeding of Colloidal Au Nanoparticle Solutions Chem. Mater., Vol. 12, No. 2, 2000 313

the statistical mean differs from the median value of

the set. Likewise, the proximity of the median and mean
values in small colloidal particle sets demonstrate that
in a uniform particle set these two values should be
close. (ii) The standard deviation (SD) of the axial
dimensions also climbs very quickly. Once the rods are
dropped from consideration, the SD for the remaining
population drops dramatically and the median and
mean values come into close agreement. (iii) Neither the
12 nm or the next larger colloid shows evidence of
colloids with high G; the dominant population is spheri-
cal. However, the minor axis of the rods is substantially
smaller than the minor axis of spheres in the same
batch. Thus, the growth of 12-nm seed particles into
rods must be a process that is not simply limited to an
accelerated growth in one dimension but a reduced
growth rate in the other two axial directions. Both these
effects could derive from solution-phase transport/
mixing phenomena causing a reduced flux of Au to the
sides of the forming particle relative to the ends.
Alternatively, a small population of the seed particles
Figure 7. Enlarged TEM image of fourth iteration NH2OH- might exhibit substantially different properties, such as
seeded colloidal Au (panel E in Figure 6). surface adsorbates, crystal faces, or lattice structure.
Conclusions
Particle seeding appears to be a valuable addition to
the synthetic repertoire for colloidal Au, both for the
production of large particles with improved monodis-
persity and for fabrication of high aspect ratio colloidal
Au rods. Both boiling citrate and room temperature
hydroxylamine served effectively as reductants for 2.6-
and 12-nm-diameter colloidal Au seeds. For hydroxy-
lamine, iterative seeding can be used to grow (from 12-
nm-diameter seeds) particles with 100-200-nm major
axes. In our laboratories, the protocols reported herein
have completely supplanted the more traditional ap-
proaches to syntheses of large-diameter colloidal Au
nanoparticles; accordingly, these methods may be of
general utility.
Acknowledgment. Support from NSF (CHE-
Figure 8. Histogram showing the number of particles exhib- 9256692, CHE-9627338), NIH (GM55312-01), and the
iting a given G for the Au colloid depicted in panel D of Figure Beckman Foundation is gratefully acknowledged. Ac-
6. The same data are replotted in the inset, with each particle
number based on sorted values of G. knowledgment is also made to the Electron Microscopy
Facility for the Life Sciences in the Biotechnology
G value, amplifies this point. The transition from G < Institute at The Pennsylvania State University.
1.5 to G >1.5 is extremely abrupt, with a 2 order of
Supporting Information Available: TEM image of 2.6-
magnitude difference in particle populations. Equally nm diameter seeds, and optical spectra as well as synthetic
revealing is the inset to Figure 8, a replotting of the protocols for the particles described in Figure 6 and Table 2.
same data based on sorted G values: nearly 3000 of the This material is available free of charge via the Internet at
3300 particles have a G close to 1, and the remainder http://pubs.acs.org.
have G values from 2 to as high as 8. CM980065P
Table 2 gives size statistics for both particle shapes
individually and for the total set of particles,31 from (31) For the smaller diameter particle sets, particularly A, assign-
which several key points are revealed. (i) Comparison ment of the particles to the categories of rod or sphere is somewhat
arbitrary, given the narrow size distribution and low ellipticiity.
of the median and mean values is provided to show that Accordingly, a G value of 1.2 was used as the cutoff for A and 1.3 was
for samples containing a bimodal sample distribution, used for B.