Gold Nanoparticles: Assembly, Supramolecular Chemistry, Quantum-Size-Related Properties, and Applications toward Biology, Catalysis, and Nanotechnology Marie-Christine Daniel and Didier Astruc* Molecular Nanosciences and Catalysis Group, LCOO, UMR CNRS No. 5802, Universite �� Bordeaux I, 33405 Talence Cedex, France Received August 6, 2003 Contents 1. Historic Introduction 293 2. General Background: Quantum Size Effect and Single-Electron Transitions 294 3. Synthesis and Assembly 296 3.1. Citrate Reduction 296 3.2. The Brust-Schiffrin Method: Two-Phase Synthesis and Stabilization by Thiols 296 3.3. Other Sulfur Ligands 297 3.4. Other Ligands 298 3.4.1. Phosphine, Phosphine Oxide, Amine, and Carboxylate Ligands 298 3.4.2. Isocyanide 298 3.4.3. Acetone 298 3.4.4. Iodine 298 3.5. Microemulsion, Reversed Micelles, Surfactants, Membranes, and Polyelectrolytes 298 3.6. Seeding Growth 298 3.7. Physical Methods: Photochemistry (UV, Near-IR), Sonochemistry, Radiolysis, and Thermolysis 298 3.8. Solubilization in Fluorous and Aqueous Media 299 3.9. Characterization Techniques 300 3.10. Bimetallic Nanoparticles 303 3.11. Polymers 304 3.12. Dendrimers 307 3.13. Surfaces, Films, Silica, and Other AuNP Materials 308 4. Physical Properties 312 4.1. The Surface Plasmon Band (SPB) 312 4.2. Fluorescence 314 4.3. Electrochemistry 315 4.4. Electronic Properties Using Other Physical Methods 315 5. Chemical, Supramolecular, and Recognition Properties 317 5.1. Reactions of Thiolate-Stabilized AuNPs 317 5.2. Supramolecular Chemistry 318 5.3. Molecular Recognition 319 5.3.1. Redox Recognition Using Functionalized AuNPs as Exoreceptors 319 5.3.2. Miscellaneous Recognition and Sensors 320 6. Biology 321 6.1. DNA-AuNPs Assemblies and Sensors 321 6.2. AuNP-Enhanced Immuno-Sensing 323 6.3. AuNP Sugar Sensors 323 6.4. Other AuNP Bioconjugates: Peptides, Lipids, Enzymes, Drugs, and Viruses 324 6.5. AuNP Biosynthesis 325 7. Catalysis 325 7.1. Catalysis of CO Oxidation 325 7.2. Electrochemical Redox Catalysis of CO and CH3OH Oxidation and O2 Reduction 326 7.3. Catalysis of Hydrogenation of Unsaturated Substrates 326 7.4. Catalysis by Functional Thiolate-Stabilized AuNPs 326 7.5. Other Types of Catalysis 327 8. Nonlinear Optics (NLO) 327 9. Miscellaneous Applications 328 10. Conclusion and Perspectives 329 11. Acknowledgment 329 12. Abbreviations 329 13. References 330 1. Historic Introduction Although gold is the subject of one of the most ancient themes of investigation in science, its renais- sance now leads to an exponentially increasing number of publications, especially in the context of emerging nanoscience and nanotechnology with nano- particles and self-assembled monolayers (SAMs). We will limit the present review to gold nanoparticles (AuNPs), also called gold colloids. AuNPs are the most stable metal nanoparticles, and they present fascinating aspects such as their assembly of multiple types involving materials science, the behavior of the individual particles, size-related electronic, magnetic and optical properties (quantum size effect), and their applications to catalysis and biology. Their promises are in these fields as well as in the bottom-up approach of nanotechnology, and they will be key materials and building block in the 21st century. Whereas the extraction of gold started in the 5th millennium B.C. near Varna (Bulgaria) and reached 10 tons per year in Egypt around 1200-1300 B.C. when the marvelous statue of Touthankamon was constructed, it is probable that ���soluble��� gold ap- peared around the 5th or 4th century B.C. in Egypt and China. In antiquity, materials were used in an ecological sense for both aesthetic and curative purposes. Colloidal gold was used to make ruby glass 293 Chem. Rev. 2004, 104, 293-346 10.1021/cr030698+ CCC: $48.50 �� 2004 American Chemical Society Published on Web 12/20/2003

and for coloring ceramics, and these applications are still continuing now. Perhaps the most famous ex- ample is the Lycurgus Cup that was manufactured in the 5th to 4th century B.C. It is ruby red in transmitted light and green in reflected light, due to the presence of gold colloids. The reputation of soluble gold until the Middle Ages was to disclose fabulous curative powers for various diseases, such as heart and venereal problems, dysentery, epilepsy, and tumors, and for diagnosis of syphilis. This is well detailed in what is considered as the first book on colloidal gold, published by the philosopher and medical doctor Francisci Antonii in 1618.1 This book includes considerable information on the formation of colloidal gold sols and their medical uses, including successful practical cases. In 1676, the German chemist Johann Kunckels published another book,2a whose chapter 7 concerned ���drinkable gold that contains metallic gold in a neutral, slightly pink solution that exert curative properties for several diseases���. He concluded, well before Michael Faraday (vide infra), that ���gold must be present in such a degree of communition that it is not visible to the human eye���. A colorant in glasses, ���Purple of Cas- sius���, is a colloid resulting from the heterocoagulation of gold particles and tin dioxide, and it was popular in the 17th century.2b A complete treatise on colloidal gold was published in 1718 by Hans Heinrich Helcher.3 In this treatise, this philosopher and doctor stated that the use of boiled starch in its drinkable gold preparation noticeably enhanced its stability. These ideas were common in the 18th century, as indicated in a French dictionary, dated 1769,4 under the heading ���or potable���, where it was said that ���drinkable gold contained gold in its elementary form but under extreme sub-division suspended in a liquid���. In 1794, Mrs. Fuhlame reported in a book5 that she had dyed silk with colloidal gold. In 1818, Jeremias Benjamin Richters suggested an explana- tion for the differences in color shown by various preparation of drinkable gold:6 pink or purple solu- tions contain gold in the finest degree of subdivision, whereas yellow solutions are found when the fine particles have aggregated. In 1857, Faraday reported the formation of deep- red solutions of colloidal gold by reduction of an aqueous solution of chloroaurate (AuCl4-) using phosphorus in CS2 (a two-phase system) in a well- known work. He investigated the optical properties of thin films prepared from dried colloidal solutions and observed reversible color changes of the films upon mechanical compression (from bluish-purple to green upon pressurizing).7 The term ���colloid��� (from the French, colle) was coined shortly thereafter by Graham, in 1861.8 Although the major use of gold colloids in medicine in the Middle Ages was perhaps for the diagnosis of syphilis, a method which re- mained in use until the 20th century, the test is not completely reliable.9-11 In the 20th century, various methods for the preparation of gold colloids were reported and reviewed.11-17 In the past decade, gold colloids have been the subject of a considerably increased number of books and reviews,15-44 especially after the break- throughs reported by Schmid17,19,21 and Brust et al.22,27 The subject is now so intensively investigated, due to fundamental and applied aspects relevant to the quantum size effect, that a majority of the references reported in the present review article have appeared in the 21st century. Readers interested in nano- particles in general can consult the excellent books cited in refs 15, 24, 28, 33, and 43. The book by Hayat, published in 1989,14 essentially deals with biological aspects and imaging of AuNPs. 2. General Background: Quantum Size Effect and Single-Electron Transitions Physicists predicted that nanoparticles in the diameter range 1-10 nm (intermediate between the size of small molecules and that of bulk metal) would display electronic structures, reflecting the electronic band structure of the nanoparticles, owing to quan- tum-mechanical rules.29 The resulting physical prop- erties are neither those of bulk metal nor those of molecular compounds, but they strongly depend on Marie-Christine Daniel was born in Vannes, France. She graduated from the University of Rennes (France). She is now finishing her Ph.D. on exoreceptors at the Bordeaux 1 University in the research group of Professor Didier Astruc. Her doctoral research is concerned with the recognition of anions of biological interest using functionnalized gold nanoparticles and redox-active metallodendrimers. Didier Astruc is Professor of Chemistry at the University Bordeaux I and has been a Senior Member of the Institut Universitaire de France since 1995. He studied in Rennes (thesis with R. Dabard), and then did his postdoctoral research at MIT with R. R. Schrock. He is the author of Electron Transfer and Radical Processes in Transition-Metal Chemistry (VCH, 1995, prefaced by Henry Taube) and Chimie Organome ��tallique (EDP Science, 2000 Spanish version in 2003). His research interests are in organometallic chemistry at the interface with nanosciences, including sensing, catalysis, and molecular electronics. 294 Chemical Reviews, 2004, Vol. 104, No. 1 Daniel and Astruc

the particle size, interparticle distance, nature of the protecting organic shell, and shape of the nano- particles.27 The few ���last metallic electrons��� are used for tunneling processes between neighboring par- ticles, an effect that can be detected by impedance measurements that distinguish intra- and intermo- lecular processes. The quantum size effect is involved when the de Broglie wavelength of the valence electrons is of the same order as the size of the particle itself. Then, the particles behave electroni- cally as zero-dimensional quantum dots (or quantum boxes) relevant to quantum-mechanical rules. Freely mobile electrons are trapped in such metal boxes and show a characteristic collective oscillation frequency of the plasma resonance, giving rise to the so-called plasmon resonance band (PRB) observed near 530 nm in the 5-20-nm-diameter range. In nanoparticles, there is a gap between the valence band and the conduction band, unlike in bulk metals. The size- induced metal-insulator transition, described in 1988, is observed if the metal particle is small enough (about 20 nm) that size-dependent quantization ef- fects occur. Then, standing electron waves with discrete energy levels are formed. Single-electron transitions occur between a tip and a nanoparticle, causing the observation of so-called Coulomb block- ades if the electrostatic energy, Eel ) e2/2C, is larger than the thermal energy, ET ) kT. The capacitance C becomes smaller with smaller particles. This means that single-electron transitions can be observed at a given temperature only if C is very small, i.e., for nanoparticles since they are small enough (C 10-18 F). Large variations of electrical and optical proper- ties are observed when the energy level spacing exceeds the temperature, and this flexibility is of great practical interest for applications (transistors, switches, electrometers, oscillators, biosensors, cataly- sis).32-38 For instance, single-electron tunneling re- lated to the electrical resistance of a single rod- shaped molecule provided a value of 18 ( 12 M��� for self-assembled monolayers on gold (1,1,1) substrate used to tether AuNPs deposited from a cluster beam.32 The transition from metal-like capacitive charging to redox-like charging was observed with alkanethiolate-gold nanoparticles of low dispersity in an electrochemical setup for Coulomb staircase experiments.39,40 Indeed, it was initially indicated that these AuNPs could accommodate 10 redox states.39a In a subsequent paper published in 2003, it was shown that lower temperatures enhance the resolution of quantized double-layer charging peaks in differential pulse voltammetry (DPV) observations. This led to the resolution of 13 peaks in CH2Cl2 at 263 K for Au140 particles.39b At the same time, however, a publication by Quinn���s group revealed remarkably well-resolved DPV of analogous Au147 particles, showing 15 evenly spaced peaks at room temperature (295 K) corresponding to 15 oxidation states (Figure 1). It was also anticipated that, the number of observable charge states being limited by the size of the available potential window, additional peaks should be observed in controlled atmosphere and reduced temperature conditions.40 Thus, AuNPs behave as other delocalized redox molecules, disclos- ing redox cascades that are well known in inorganic and organometallic electrochemistry for other transi- tion metal clusters and bi-sandwich complexes. The pioneering work by Schmid and co-workers on well-defined phosphine-stabilized gold clusters showed the properties of quantum-dot particles for the first time.30 The number of atoms in these gold clusters is based on the dense packing of atoms taken as spheres, each atom being surrounded by 12 nearest neighbors. Thus, the smallest cluster contains 13 atoms, and the following layers contain 10n2 + 2 atoms, n being the layer number. For instance, the second layer contains 42 atoms, which leads to a total of 55 atoms for a gold cluster, and the compound [Au55(PPh3)12Cl6] has been well characterized by Schmid���s group. Recently, spectroscopic data have revealed discrete energy level spacings of 170 meV that can be attributed to the Au55 core.30c Larger clusters containing, respectively, 147, 309, 561, 923, 1415, or 2057 (n ) 3-8) atoms have been isolated.30,31 Discrete organogold clusters are also well known with small numbers of atoms and various geometries, and they will not be reviewed here.41,42 Large ones form a fuzzy frontier between clusters and colloids (AuNPs), the latter being defined by some dispersity material- ized by a histogram determined using transmission electron microscopy (TEM) data. Despite the considerable variety of contributions, we will focus first on synthesis, stabilization, and various types of assemblies, and then on physical properties and on chemical, supramolecular, and sensor properties, and finally on applications to biochemistry, catalysis, and nonlinear optical proper- ties before concluding on the perspectives of AuNPs in nanosciences and nanotechnology. Many publica- tions involve two or even sometimes several of these topics. Thus our classification is arbitrary, but the reader will often better understand the spirit of each paper from its title given in the reference section. Figure 1. Differential pulse voltammetry (DPV) responses for AuNP solutions measured at a Pt microelectrode (upper) as-prepared 177 ��M hexanethiol-capped Au147 showing 15 high-resolution quantized double-layer charg- ing (QDL) peaks and (lower) 170 ��M hexanethiol-capped Au38 showing a HOMO-LUMO gap. It can be seen that the as-prepared solution contains a residual fraction of Au38 that smears out the charging response in E regions where QDL peaks overlap. The electrode potential scanned nega- tive to positive. Reprinted with permission from ref 40 (Quinn���s group). Copyright 2003 American Chemical So- ciety. Gold Nanoparticles Chemical Reviews, 2004, Vol. 104, No. 1 295

3. Synthesis and Assembly 3.1. Citrate Reduction Among the conventional methods of synthesis of AuNPs by reduction of gold(III) derivatives, the most popular one for a long time has been that using citrate reduction of HAuCl4 in water, which was introduced by Turkevitch in 1951.12 It leads to AuNPs of ca. 20 nm. In an early effort, reported in 1973 by Frens,13 to obtain AuNPs of prechosen size (between 16 and 147 nm) via their controlled formation, a method was proposed where the ratio between the reducing/stabilizing agents (the trisodium citrate-to- gold ratio) was varied. This method is very often used even now when a rather loose shell of ligands is required around the gold core in order to prepare a precursor to valuable AuNP-based materials. Re- cently, a practical preparation of sodium 3-mercap- topropionate-stabilized AuNPs was reported in which simultaneous addition of citrate salt and an am- phiphile surfactant was adopted the size could be controlled by varying the stabilizer/gold ratio (Figure 2).44 3.2. The Brust-Schiffrin Method: Two-Phase Synthesis and Stabilization by Thiols Schmid���s cluster [Au55(PPh3)12Cl6], reported in 1981, long remained unique with its narrow dispersity (1.4 ( 0.4 nm) for the study of a quantum-dot nanoma- terial, despite its delicate synthesis.45 The stabiliza- tion of AuNPs with alkanethiols was first reported in 1993 by Mulvaney and Giersig, who showed the possibility of using thiols of different chain lengths and their analysis.46a The Brust-Schiffrin method for AuNP synthesis, published in 1994, has had a considerable impact on the overall field in less than a decade, because it allowed the facile synthesis of thermally stable and air-stable AuNPs of reduced dispersity and controlled size for the first time (ranging in diameter between 1.5 and 5.2 nm). Indeed, these AuNPs can be repeatedly isolated and redissolved in common organic solvents without irreversible aggregation or decomposition, and they can be easily handled and functionalized just as stable organic and molecular compounds. The tech- nique of synthesis is inspired by Faraday���s two-phase system7 and uses the thiol ligands that strongly bind gold due to the soft character of both Au and S.47 AuCl4- is transferred to toluene using tetraoctylam- monium bromide as the phase-transfer reagent and reduced by NaBH4 in the presence of dodecanethiol (Figure 3).47a The organic phase changes color from orange to deep brown within a few seconds upon addition of NaBH4: The TEM photographs showed that the diameters were in the range 1-3 nm, with a maximum in the particle size distribution at 2.0-2.5 nm, with a preponderance of cuboctahedral and icosahedral struc- tures. Larger thiol/gold mole ratios give smaller average core sizes, and fast reductant addition and cooled solutions produced smaller, more monodis- perse particles. A higher abundance of small core sizes (e2 nm) is obtained by quenching the reaction immediately following reduction or by using sterically bulky ligands.48-50 Brust et al. extended this synthe- sis to p-mercaptophenol-stabilized AuNPs in a single- phase system,47b which opened an avenue to the synthesis of AuNPs stabilized by a variety of func- tional thiol ligands.47,48 Subsequently, many publica- tions appeared describing the use of the Brust- Schiffrin procedure for the synthesis of other stable AuNPs, also sometimes called monolayer-protected clusters (MPCs), of this kind that contained func- tional thiols.49-53 The proportion thiol:AuCl4- used in the synthesis controls the size of the AuNPs (for instance, a 1:6 ratio leads to the maximum average core diameter of 5.2 nm, i.e., ca. 2951 Au atoms and ca. 371 thiolate ligands core diameter dispersity of ��� (10%). Murray et al. reported and studied the ���place exchange��� of a controlled proportion of thiol ligands by various functional thiols52 (Figures 4 and 5) and the subsequent reactions of these functional AuNPs.50,52 Schiffrin reported the purification of dodecanethiol-stabilized AuNPs from tetraoctylam- Figure 2. Preparation procedure of anionic mercapto- ligand-stabilized AuNPs in water. Reprinted with permis- sion from ref 44 (Kunitake���s group). Copyright 1999 Elsevier. Figure 3. Formation of AuNPs coated with organic shells by reduction of AuIII compounds in the presence of thiols. Reprinted with permission from ref 73 (Crooks���s group). Copyright 2001 Royal Society of Chemistry. AuCl4-(aq) + N(C8H17)4+(C6H5Me) f N(C8H17)4+AuCl4-(C6H5Me) mAuCl4-(C6H5Me) + nC12H25SH(C6H5Me) + 3m e- f 4m Cl-(aq) + [Aum(C12H25SH)n](C6H5Me) 296 Chemical Reviews, 2004, Vol. 104, No. 1 Daniel and Astruc

monium impurities by Soxhlet extraction.54 The influence of nonionic surfactant polyoxoethylene(20) sorbitan monolaurate (Tween 20) on surface modifi- cation of AuNPs was studied with mercaptoalkanoic acids.55 Digestive ripening, i.e., heating a colloidal suspension near the boiling point in the presence of alkanethiols (for instance, 138 ��C for 2 min, followed by 5 h at 110 ��C), significantly reduced the average particle size and polydispersity in a convenient and efficient way. This technique also led to the formation of 2D and 3D superlattices,56,57 a subject of intense investigation (see also section 3.13 on materials).58-63 For instance, AuNPs obtained using acid-facilitated transfer are free of tetraalkylammonium impurity, are remarkably monodisperse, and form crystalline superstructures.63a The truncated icosahedron struc- ture is formed in growth conditions in which the equilibrium shape is achieved.63b Molecular dynamics simulations showed that AuNPs with 1157 Au atoms attained an icosahedral structure upon freezing.63c A single-toluene phase method was also reported whereby the ammonium salt-stabilized AuNPs were synthesized, followed by an exchange reaction with dodecanethiol.58 Superhydride64a and hexadecyl- aniline64b (inter alia) have been used as alternative reagents to NaBH4 for the reduction of gold(III) in the synthesis of thiol-stabilized AuNPs. Shape sepa- ration of suspended AuNPs by size-exclusion chro- matography was monitored by examining the 3D chromatograms obtained by employing a diode-array detection system.65 3.3. Other Sulfur Ligands Other sulfur-containing ligands,67-70 such as xan- thates66 and disulfides,67-69 di-70a and trithiols,70b and resorcinarene tetrathiols,70d have been used to sta- bilize AuNPs. Disulfides are not as good stabilizing agents as thiols,67-70 which is eventually useful for catalysis.70 Similarly, thioethers do not bind AuNPs strongly,71 but the use of polythioethers by Rhein- hout���s group astutely circumvented this problem.72a Tetradentate thiethers have also been used to revers- ibly form AuNP assemblies.72b On the other hand, oxidation of thiol-stabilized AuNPs by iodine pro- vokes their decomposition to gold iodide with forma- tion of disulfides, which led Crooks to form polycy- clodextrin hollow spheres by templating AuNPs.73 Figure 4. General scheme for the ligand-exchange reac- tion between alkanethiol-AuNPs of the Brust type and various functionalized thiols. Figure 5. Ligand substitution reactions (CH2Cl2, 2 d, room temperature) for the syntheses of the AuNPs containing mixed dodecanethiol and (amidoferrocenyl) alkanethiol-type ligands with variation of the chain length (C11 vs C6) and ring structure of the ferrocenyl motif (Cp, Cp*, C5H4COMe). Reprinted with permission from ref 140 (Astruc���s group). Copyright 2002 American Chemical Society. Gold Nanoparticles Chemical Reviews, 2004, Vol. 104, No. 1 297

3.4. Other Ligands 3.4.1. Phosphine, Phosphine Oxide, Amine, and Carboxylate Ligands The Brust biphasic method of synthesis was ap- plied to PPh3 in order to improve the synthesis of Schmid���s cluster [Au55(PPh3)12Cl6], using HAuCl4��� 3H2O and N(C8H15)4Br in a water-toluene mixture to which PPh3 and then NaBH4 were added. It was estimated that the cluster synthesized in this way had the formula [Au101(PPh3)21Cl5] and contained 3.7 mass percent of [Au(PPh3)Cl] as an impurity.74a Thermolysis of [AuI(C13H27COO)(PPh3)] at 180 ��C under N2 yielded monodispersed AuNPs capped by myristate and a small amount of PPh3 ligands the AuNP diameter increased with reaction time, from 12 nm for 1 h to 28 nm for 10 h, and with increasing temperature (42 nm for 5 h at 200 ��C).74b Various other gold complexes, in particular gold(I) amine complexes, have been used as precursors for the synthesis of amine-stabilized AuNPs.75,76a,b Reduction of AuIVCl4 by NaBH4 in a mixture of tri-n-octylphos- phine oxide (TOPO) and octadecylamine (1:0.57 molar ratio) at 190 ��C resulted in the controlled growth of spherical AuNPs (8.59 ( 1.09 nm diameter) that are stable for months in toluene and were manipulated into crystals and 2D arrays (Figure 6).76c Capping aqueous AuNPs with the amino acid lysine stabilizes the AuNPs in solution electrostatically and renders them air-stable and water-dispersible, a finding that is promising toward biologically relevant research.76d Efficient synthesis of stable AuNPs by reaction of AuCl4- ions with the alkalothermophilic actinomycete Thermomonospora sp. has been described.76e 3.4.2. Isocyanide Aryl isocyanide thin films have attracted some attention, due to their potential application as mo- lecular wires,76f and 1,4-diisocyanide-AuNP forms large aggregate superstructures that have been examined by IR and Raman spectroscopy, showing bonding to the AuNP core via the carbon lone pair.76g,h 3.4.3. Acetone Pure Au0NPs, obtained by replacement of citrate by acetone, were shown to be stable against attack by BH4- or HCl.76i 3.4.4. Iodine Iodine adsorption was shown to displace citrate ions from AuNPs, leading to superstructures that are also formed upon addition of KI.76j 3.5. Microemulsion, Reversed Micelles, Surfactants, Membranes, and Polyelectrolytes The use of microemulsions,77 copolymer micelles,78 reversed micelles,77 surfactant, membranes, and other amphiphiles is a significant research field for the synthesis38 of stabilized AuNPs in the presence or in the absence of thiol ligands.77-95 The syntheses involve a two-phase system with a surfactant that causes the formation of the microemulsion or the micelle maintaining a favorable microenvironment, together with the extraction of metal ions from the aqueous phase to the organic phase. This is an advantage over the conventional two-phase system. This dual role of the surfactant and the interaction between the thiol and the AuNP surface control the growth and stabilization of the AuNP or nanocrystal. The narrow size distribution allows the ordering of the particles into a 2D hexagonal close-packed array. AuNP sizes of the order of 4 nm diameter have been found.79 Polyelectrolytes have also been extensively used for the synthesis of AuNPs (Figure 7).94,98-102 The polyelectrolyte coating of carboxylic acid-deriva- tized AuNPs with diameters less than 10 nm has been achieved by electrostatic self-assembly of op- positely charged polyelectrolytes.102b 3.6. Seeding Growth The seeding-growth procedure is another popular technique that has been used for a century. Recent studies have successfully led to control of the size distribution (typically 10-15%) in the range 5-40 nm, whereas the sizes can be manipulated by varying the ratio of seed to metal salt (Figure 8).103-105 The step-by-step particle enlargement is more effective than a one-step seeding method to avoid secondary nucleation.87a Gold nanorods have been conveniently fabricated using the seeding-growth method.87b 3.7. Physical Methods: Photochemistry (UV, Near-IR), Sonochemistry, Radiolysis, and Thermolysis UV irradiation is another parameter that can improve the quality of the AuNPs,86,104,105 including when it is used in synergy with micelles86 or seeds.104 Near-IR laser irradiation provokes an enormous size growth of thiol-stabilized AuNPs.106 The presence of an ultrasonic field (200 kHz) allowed the control of the rate of AuCl4- reduction in an aqueous solution containing only a small amount of 2-propanol and the Figure 6. 2D lattice of octadecylamine/TOPO-capped AuNPs spontaneously formed when the latter are deposited on a copper grid bar ) 20 nm. (Inset) Scanning electron microscopy (SEM) image of a cubic colloidal crystal pre- pared from octadecylamine/TOPO-capped AuNPs (190 ��C) bar ) 80 ��m. Reprinted with permission from ref 76c (O���Brien���s group). Copyright 2000 The Royal Society of Chemistry. 298 Chemical Reviews, 2004, Vol. 104, No. 1 Daniel and Astruc

sizes of the formed AuNPs by using parameters such as the temperature of the solution, the intensity of the ultrasound, and the positioning of the reac- tor.107,108 Sonochemistry was also used for the syn- thesis of AuNPs within the pores of silica111-113 and for the synthesis of Au/Pd bimetallic particles.114 Radiolysis has been used to control the AuNP size115a or to synthesize them in the presence of specific radicals,115b and the mechanism of AuNP formation upon ��-irradiation has been carefully examined (Fig- ure 9).116 AuNPs have been fabricated via decomposition of [AuCl(PPh3)] upon reduction in a monolayer at the gas/liquid interface.117 The thermolysis of [C14H29- Me3N][Au(SC12H25)2] at 180 ��C for 5 h under N2 produced alkyl-groups-passivated AuNPs of 26 nm.118a Thermolysis of crude preparations of Brust���s AuNPs without removing the phase-transfer reagent, tet- raoctylammonium bromide, to 150-25 ��C led to an increase of the particle sizes to 3.4-9.7 nm, and this size evolution was discussed on the basis of a thermodynamic model. The heat-treated AuNPs formed 2D superlattices with hexagonal packing. The conformation of the alkanethiol is all-trans, and these ligands interpenetrate each other (Figure 10).118b Laser photolysis has been used to form AuNPs in block copolymer micelles.119 Laser ablation is another technique of AuNP synthesis that has been used under various conditions whereby size control can be induced by the laser.120-122 The evolution of thiol- stabilized AuNPs has been induced by and observed upon heating.123-126,135 Structural changes of spheri- cal aggregates composed of mercaptoacetate-stabi- lized AuNPs suspended in water were monitored by maintaining the spheroid suspension at a constant temperature, ranging from 65 to 91 ��C, for 2-12 h. The spheroid diameter was reduced to almost 70% of the original size, due to an irreversible ���coagula- tive��� transition resulting from fusion among the nanocolloids in spheroids.126b Morphology changes of AuNPs were also shown during sintering.126c Sput- tering AuNPs by single ions and clusters was shown to eject AuNPs.178 3.8. Solubilization in Fluorous and Aqueous Media AuNPs stabilized by perfluorodecanethiol or 1H,1H,2H,2H-perfluorooctathiol, with an average Figure 7. Schematic diagram illustrating the layer-by- layer polymer deposition process applied to AuNPs. Re- printed with permission from ref 98 (Caruso���s group). Copyright 2001 American Chemical Society. Figure 8. TEM image of larger gold particles prepared from seed: (a) 5.5 ( 0.6, (b) 8.0 ( 0.8, (c) 17 ( 2.5, and (d) 37 ( 5 nm after separation of rods. The 5.5 ( 0.6-nm AuNPs were extracted into toluene after thiol capping for TEM in order to remove excess surfactant. The other particles were separated from excess surfactant by cen- trifugation. Reprinted with permission from ref 89 (Mur- phy���s group). Copyright 2001 American Chemical Society. Gold Nanoparticles Chemical Reviews, 2004, Vol. 104, No. 1 299

diameter of 2.4-2.6 nm, were prepared by reduction of HAuCl4 by NaBH4 (dropwise addition) in ethanol and were soluble only in fluorocarbon media.127 Special emphasis has been placed on the synthesis of stable water-soluble thiol-stabilized AuNPs128-134 using thiols containing poly(ethylene oxide) chains128-130a (Figure 11) or carboxylate modification (Figure 12).130b,134 Poly(N-vinyl-2-pyrrolidone) (PVP) is the polymer of choice for the stabilization in water of AuNPs prepared by reduction of HAuCl4 (see the section on polymers). 3.9. Characterization Techniques The most common characterization technique is high-resolution transmission electron microscopy (HRTEM), which gives a photograph of the gold core of the AuNPs,46 but the core dimensions can also be determined using scanning tunneling microscopy (STM), atomic force microscopy (AFM), small-angle X-ray scattering (SAXS),50,137a laser desorption- ionization mass spectrometry (LDI-MS),137b-139 and X-ray diffraction.136 A detailed high-resolution study of the AuNP shape using HRTEM, reported by Brust et al., revealed that the truncated cuboctahedron predominated, and that decahedra, dodecahedra and icosahedra were also present in the same preparation of alkanethiol-stabilized AuNPs.47 The histogram providing the size distribution of these cores gives crucial information on the dispersity of the sample that is usually obtained from TEM pictures.47 The mean diameter, d, of the cores allows determination of the mean number of gold atoms, NAu, in the cores:47 NAu ) 4��(d/2)3/vAu. For instance, with d ) 2.06 nm, NAu ) 269.140 From these data, the ele- mental analysis, giving the Au/S ratio, allows calcu- lation of the average number of S ligands. This number can also be deduced from X-ray photoelectron spectroscopy (XPS) or thermogravimetric analysis (TGA).50 Figure 9. Taping-mode AFM images of the species formed in a solution irradiated with �� rays (1.5 kGy) and then deposited on highly ordered pyrolytic graphite (HOPG) 30 days after the irradiation and dried under a mild N2 stream for visualization. The solution contains 10-3 mol���L-1 AuIII and poly(vinyl alcohol) but no alcohol. (A) Height-mode (z range 80.0 nm) image and (B) phase (z range 42.2��) image, run simultaneously on the same area of the sample. (C, D) Close-up images showing the same area as in (A) and (B), respectively. Reprinted from ref 116 (Belloni���s group) by permission of The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS). Copyright 1998. Figure 10. UV-vis spectra (A) and TEM images and size distributions (B) of (a) [AuCl4]- before reduction dode- canethiol-AuNPs (b) as prepared and after heat treatment at (c) 150, (d) 190, and (e) 230 ��C and (f) octadecanethiol- AuNPs heat-treated at 250 ��C. Reprinted with permission from ref 188b (Miyake���s group). Copyright 2003 American Chemical Society. 300 Chemical Reviews, 2004, Vol. 104, No. 1 Daniel and Astruc

The oxidation state of the gold atoms of the core has been examined by Brust et al in their seminal article using X-ray photoelectron spectra that showed the binding energies of the doublet for Au 4f7/2 (83.8 eV) and Au 4f5/2 (87.5 eV) characteristic of Au0. No band was found for AuI at 84.9 eV, although one-third of the gold atoms are located at the surface and bonded to thiols for 2.0-2.5 nm sized particle cores. On this basis, Brust et al. suggested that the gold- thiol bond does not have the character of gold sulfide.46b This matter of a thiol vs thiolate bond to the gold core atoms, however, has been debated. For instance, it was suggested that a high coverage of gold cores by thiolate ligands139 was due to large ligand/Au binding ratios on core edges and vertexes (Figure 13), in accord with theoretical calculations.140 Moreover, it was reported that thermolysis of the thiolate-stabilized AuNPs produces only the corre- sponding disulfide. The absence of thiol by thermal desorption mass spectrometry was considered to be evidence that the chemisorbed ligand consisted of an alkanethiolate (not thiol) fragment. This would mean that H2 is produced during the reductive synthesis from thiols, but this formation has never been detected. Theoretical calculations suggested the for- mation of disulfides when the number of thiol mol- ecules around a AuNP was enough to saturate the flat planes, whereas thiolate behavior was observed when the sulfur atoms were not enough.139a This finding was reported139 to corroborate the observation of a S-S distance of 2.32 �� from grazing incidence X-ray in self-assembly of n-alkanethiols on a (1,1,1) gold crystal surface.139b Brust et al. recently provided 1H NMR evidence for intact thiols adsorbed on AuNPs. They showed that the loss of hydrogen could be prevented to some extent as long as there is no easy reaction path for hydrogen removal.140 X-ray diffraction also demonstrated the striking tendency of thiolates-AuNPs to spontaneously form highly ordered superlattices141-144 with periodicity extending to three dimensions up to several tens of micrometers.143 These superlattices were obtained upon slow evaporation of the organic solvent or even water on a suitable surface.144b Such self-organized superlattices of AuNPs on highly ordered pyrolytic graphite (HOPG) are also observable by STM (Figure 14).27 Monodispersity is a very important criterion for the formation of ordered superlattices.145a When the gas phase at the gas-suspension interface of a synthetic medium leading to sodium mercaptosucci- nate-AuNPs contained nonpolar organic molecules, spherical AuNPs formed. On the other hand, when the gas phase contained polar organic vapors such as MeCN or CHCl3, irregular-shaped AuNPs formed.145b Langmuir-Blodgett (LB) films of Schmid���s Au55 cluster were characterized by STM, Brewter angle microscopy (BAM), and scanning force microscopy (SFM). These techniques showed that the cluster formed monolayers, as indicated by the surface pressure-area (��-A) isotherms and the area-time (A-t) isobars between 20 and 30 ��C and 15-30 nN/m of surface pressure, and the cluster size could be estimated from the ��-A isotherms as 2.17 nm (calcd 2.1 nm).19 The Au55 cluster has also been studied by Mo ��ssbauer spectroscopy, extended X-ray absorption fine structure (EXAFS), electron spectroscopy for chemical analysis (ESCA), and conductivity measure- ments. These techniques show that the Au55 particles behave like a system with a few ���last metallic electrons��� that are used for tunneling between neigh- boring clusters. This ���metallizing��� situation was observed by applying an alternating current in the 10-kHz range as for impedance measurements.146,147 Scanning tunneling spectroscopy (STS) had been used to observe Coulomb blockade in metal nano- particles. The tunneling current is induced by an Figure 11. Schematic representation of a AuNP protected by a monolayer of monohydroxy (1-mercaptoundec-11-yl) tetraethylene glycol. The hydrophobic C11 chain confers extreme stability to the cluster, while the hydrophilic tetraethylene glycol unit ensures solubility in water. Reprinted with permission from ref 129 (Brust���s group). Copyright 2002 The Royal Society of Chemistry. Figure 12. Possible combinations of H2O molecules with mercaptosuccinic acid (MSA)-capped AuNPs: one H2O molecule connected with two carbonyl groups in either (a) one MSA molecule, (b) adjoining MSA molecules on one AuNP, making a successive hydrogen-bonding network, or (c) MSA molecules from different AuNPs, the water acting as ���glue��� to join two neighboring particles. Reprinted with permission from ref 130b (Chen���s group). Copyright 1999 American Chemical Society. Gold Nanoparticles Chemical Reviews, 2004, Vol. 104, No. 1 301

applied voltage and leads to the charging of a metal particle with at least one single electron.148-150 Elec- trostatic trapping (ET) is a technique used to inves- tigate isolated nanosize metal particles. It is based on moving a polarized particle in an electric field to the point of strongest field, which is the position between two electrodes (dipped in a solution of the particles) at a distance comparable to the particle diameter.27,148,149 The UV-vis and IR spectra provide an identifica- tion of the ligand that is also confirmed by NMR spectroscopy, except that the ligand atoms close to the core give broad signals. This latter phenomenon is due to (i) spin-spin relaxational (T2) broadening (main factor), (ii) variations among the gold-sulfur bonding sites around the particle, and (iii) a gradient in the packing density of the thiolate ligands from the core region to the ligand terminus at the periphery.151-153 The NMR spectra are very informa- tive, as for all molecular compounds, for the part of the ligand remote from the core. The latter can also be more fully analyzed, if desired, after oxidative decomplexation using iodide. IR spectroscopy shows that, as in SAMs,154 the thiolate ligands of AuNPs are essentially in all-trans zigzag conformations, with 5-25% of gauche defects at both inner and terminal locations.50,152 IR and NMR spectroscopies allow, together with differential scanning calorimetry (DSC),152-155 the detection of order-disorder transitions in AuNPs in the solid state. The temperature of the transition increases with the chain length, and FTIR shows the increasing amount of gauche defects. Variable-temperature deuterium NMR in the solid state shows that the disorder, materialized by the increased proportion of gauche bonds, propagates from the chain terminus toward the middle of the chain, but not further to the ligand atom, and causes chain melting.152 Calo- rimetric measurements led to the determination of the formation enthalpy of AuNPs in a water/sodium bis(2-ethylhexyl) sulfosuccinate/n-heptane micro- emulsion. The results indicated that the energetic states and the dimensions of the AuNPs were influ- enced by the radii and concentrations of the reversed micelles.80 Capillary zone electrophoresis in acetate buffer showed that the mobility of AuNPs with a given core diameter decreased with decreasing ionic strength. At the highest ionic strength investigated (6 mmol/ L), a good linear dependence of the mobility on the reciprocal of the core radius allowed the character- ization of the size of the AuNPs.156 The AFM images of AuNPs operating in the contact mode in air at room temperature showed an attrac- tive interaction among the particles, leading to the formation of aggregates and a mean size that is a function of the size of the reverse micelle used for the synthesis. This was taken into account in terms of the formation of an adsorbed layer of surfactant molecules at the particle surface.81 Figure 13. Stable configurations of AuNPs covered with n-alkanethiol molecules. The sequence shows n ) 4 butanethiol, n ) 6 hexanethiol, n ) 8 octanethiol, n ) 10 decanethiol, n ) 12 dodecanethiol, n ) 14 butanedecanethiol, and n ) 16 hexanedecanethiol (n ) number of C atoms). Reprinted with permission from ref 139 (Jose ��-Yacama ��n���s group). Copyright 1998 Kluwer. 302 Chemical Reviews, 2004, Vol. 104, No. 1 Daniel and Astruc

The use of incoherent light experiments, performed in the vicinity of the surface plasmon resonance frequency, allowed measurement of the phase relax- ation time and nonlinear susceptibility of AuNPs of 5-40 nm.157 Surface-enhanced Raman scattering (SERS)70b and XPS made it possible to analyze the chemisorptive properties of tetrathiol ligands and indicated that surface passivation was an important factor in the dispersibility of AuNPs in nonpolar solvents.70c EXAFS allowed investigation of the size-dependent distance contraction in thiol-stabilized AuNPs, and the short metal-ligand bond found suggested a rather strong surface interaction.158 High-resolution time-of-flight mass spectroscopy analysis of alkanethiol-stabilized AuNPs allowed the assignment of the number of gold and sulfur atoms, although alkyl chains were not evident. Pure gold cluster ions of various sizes could be generated from the AuNPs in a two-laser experiment.159 Small-angle X-ray scattering, STM, and AFM were consistent with a small, monodisperse (2.4 nm diam- eter) gold core.160a The formation by physical vapor deposition and growth of AuNPs was studied by STM on TiO2 (1,1,0) surfaces.160b The surface of AuNPs was analyzed using a phase reconstruction technique in TEM, extended to simultaneous correction of spheri- cal aberration and two-fold astigmatism.160c The vacancy formation energy of AuNPs has been shown to decrease with decreasing particle size.160d 3.10. Bimetallic Nanoparticles Bimetallic nanoparticles20,37 containing gold as one of the elements have been synthesized in a variety of ways. Bimetallic AuNPs have been reported with Ag (Figure 15),161-168 Pd (Figure 16),97,161,166,167 Pt,161,167 TiO2,99 Fe,169-171 Zn,168 Cu,165,168 ZrO2,172 CdS,173,174 Fe2O3,175 and Eu.176 Although bimetallic nano- particles have been known for a long time, Schmid���s group were the first to report the synthesis of core- shell bimetallic nanoparticles, the core-shell struc- ture being demonstrated using HRTEM and energy- disperse X-ray (EDX) microanalysis. AuNPs of 18 nm diameter were covered with a Pd or Pt shell when an aqueous solution of these AuNPs was added to a solution of H2PtCl6 or H2PdCl4 and H3NOHCl. The original color of the AuNPs then changed to brown- black. Addition of p-H2NC6H4SO3Na stabilized the generated particles in the same manner as P(m-C6H4- SO3Na)3 stabilized the AuNPs. The colloids showed a metallic luster and were of uniform 35 nm diam- eter. For instance, Au/Pt particles had an average gold content of 15% atom % located at the core surrounded by Pt crystals of about 5 nm that were pregrown before being added to the Au surface.161 Stabilization of the bimetallic particles could be achieved using the Brust procedure in the presence of thiols. Such stable bimetallic particles were syn- thesized with group 10 (Pd, Pt) and group 11 (Cu, Ag, Au) metals, all containing Au as one of the two Figure 14. Nanostruture preparation from AuNPs: STM image of self-assembled superlattice of 3.5-nm gold par- ticles on a HOPG substrate. The particles are stabilized by hexanethiol. TEM micrograph of an AB2 superlattice of AuNPs having a bimodal size distribution (4.5 and 7.8 nm). The AuNPs are stabilized by decanethiol. Reprinted with permission from ref 27 (Brust���s group). Copyright 2002 Elsevier. Figure 15. Schematic illustrating the proposed inter- actions of thiocyanate ion-coated 2.8 ( 0.8- and 11.6 ( 0.8- nm-diameter AuNPs with 37.8 ( 9-nm-diameter ethylene- diaminetetraacetic acid (EDTA)-covered AgNPs. Reprinted with permission from ref 164 (Fendler���s group). Copyright 2002 The Royal Society of Chemistry. Gold Nanoparticles Chemical Reviews, 2004, Vol. 104, No. 1 303

metals, and were characterized using TEM, 1H NMR line broadening, XPS, elemental analysis, and TGA. TEM showed that Pd/Au cores are small (1.7 nm) and relatively monodisperse (average 20% dispersity), while Ag/Au cores are larger (3.2 nm), and other bimetallic particles are of intermediate size. The mole ratios of metals both in and on the surface of the bimetallic cores differed significantly from the metal: salt ratio used in the bimetallic particle synthesis.162 Partially segregated alloys indeed form easily, and more noble metals prefer the nonsurface (core) loca- tion.20 Metal galvanic exchange reactions are yet another quite facile way to synthesize stable bimetal- lic particles. This procedure relies on reactions of alkanethiolate-metal particles or other metal par- ticles (Ag, Pd, Cu) with the complexes [AuISCH2- (C6H4)CMe3] and [PdII{S(CH2)11Me}2].163,166 Au core- Ag shell and Au core-Pt shell nanoparticles have been formed using photochemically reduced phos- photungstate Keggin ions.168b Specific properties and functions of bimetallic nanoparticles will be discussed in the appropriate sections devoted to catalytic, electronic, and optical properties. 3.11. Polymers Since the report in Helcher���s treatise in 1718,3 indicating that starch stabilizes water-soluble gold particles, it has been known that such materials, recognized two centuries later as polymers, favor the isolation of AuNPs.14,179 With the considerably im- proved recent understanding of the parameters lead- ing to the stabilization of AuNPs and of their quan- tum-size-related interest, there has been a revival of activity in the field of polymer-stabilized AuNPs.33,177,180-181 The most commonly used polymers for the stabilization of AuNPs are PVP and poly- (ethylene glycol).11b,14 Although there are a variety of ways to achieve nanoparticle-polymer composites,182,183 two different approaches dominate. The first one consists of the in situ synthesis of the nanoparticles in the polymer matrix either by reduction of the metal salts dissolved in that matrix184 or by evaporation of the metals on the heated polymer surface.185 The second one, less frequently used, involves polymerization of the ma- trix around the nanoparticles.186 Recently, however, blending of premade AuNPs into a presynthesized polystyrene polymer (synthesized by anionic polym- erization) bound to a thiol group was also reported.187 Whereas the physical process involving mechanical crushing or pulverization of bulk metals and arc discharge yielded large nanoparticles with a wide size distribution, nanoparticles prepared by reduction of metal salts are small, with a narrow size distribution. This reduction processes most often use a reagent such as NaBH4188 which is added in situ, or the reductant can also be the solvent, such as an alco- hol.189,190 For instance, HAuCl4���4H2O gives stable AuNPs upon refluxing in methanol/water in the presence of PVP, even if NaOH is added subsequently to the preparation of the AuNPs.191 In poly(acryl- amide), AuCl4- cannot be reduced by alcohol, but it can be reduced by NaBH4.192 Other reductants are generated involving radiolysis, photolysis,193 or elec- trochemistry.194 The polymer-nanoparticle composite can be generated from solution (the classic mode) or can involve the immobilization by a solid polymer such as poly(acrylic acid), poly(vinyl alcohol), or PVP frequently used. Reduction of metal ions in the presence of the polymer is most often chosen because the complexation of the metal cations by the ligand atoms of the polymer is crucial before reduction. In particular, it dramatically limits the particle size.195 The most important role of the stabilizing polymer is to protect the nanoparticles from coagulation. Toshima has expressed this function quantitatively by the ���gold number���, i.e. the number of milligrams of protective polymer that just prevents 10 mL of a red gold sol from changing color to violet upon addition of 1 mL of 10% aqueous NaCl. The ���gold number��� is smaller for protective polymers that are better stabilizers.189 Core-shell PVP-stabilized Au/ Pd196 and Au/Pt197,198 nanoparticles were prepared by Yonezawa and Toshima by simultaneous alcohol reduction of the two corresponding metal salts and characterized by EXAFS. The relative order of reduc- tion in alcohol/water is seemingly controlled by the relative redox potentials, HAuCl4 being reduced more rapidly than Pd(OH)2 and PtCl62-. The AuNPs form first, and then the Pd or Pt shell forms around the AuNPs to produce the core-shell bimetallic particles. In fact, the Pd0 formed reduces AuCl4- to Au0 and thus acts as a mediator or redox catalyst for the reduction of AuCl4-, as long as any AuCl4- is left in Figure 16. Cartoon diagram of core metal galvanic exchange reactions. MPC, monolayer-protected cluster MPAC, monolayer-protected alloy cluster SC12, S(CH2)11- CH3. Reprinted with permission from ref 166a (Murray���s group). Copyright 2002 American Chemical Society. 304 Chemical Reviews, 2004, Vol. 104, No. 1 Daniel and Astruc

the solution.199,200 Attempts to synthesize Pd-core/Au- shell bimetallic particles led instead to a remarkable cluster-in-cluster structure because of this redox priority (Figure 16).201 Many ordered polymer-AuNPs are known. For instance, AuNPs in PVP were prepared by hydrazine reduction of incorporated HAuCl4. The color of the solution of HAuCl4-loaded block copolymer changed from yellow to purple, and then to bluish upon addition of a large excess of anhydrous hydrazine. The reduction can be stopped by addition of HCl, which protonates hydrazine in order to avoid coagu- lation of the AuNPs.202,203 These phenomena were also obtained with (styrene-block-ethylene oxide).204,205 The use of a diaminotriazine-functionalized diblock copolymer led to size-controlled synthesis of AuNP aggregates in solution and in thin films with thymine functionality.206 AuNPs were generated in polymeric micelles composed of amphiphilic block copoly- mers,207,208 and amphilic star-block copolymers were an ideal choice to serve as a confined reaction vessel.209 The formation of AuNPs was also controlled by using poly(methylphosphazene), whose lone pairs stabilized the AuNPs.210 Functionalized polymers have also been used as stabilizers. Poly(ethylene glycol)-based polymer was used to fabricate an AuNP sensor that reversibly binds lectin for recognition and bioassay.211 The so-called ���grafting from��� technique has been used to construct highly dense polymer brushes. For instance, several methods,212-214 includ- ing the efficient living radical polymerization (LRP), have indeed been applied to the synthesis of AuNPs coated with such a high-density polymer brush. AuNP-based nanoscale architectures could be fore- casted using this simple technique (Figure 17).214 Polymer hollow spheres have been synthesized with movable AuNPs at their interiors.215 AuNPs can serve as templates for the synthesis of conductive capsules216 (Figure 18) and for the oligomerization of L-cysteine in aqueous solution (Figure 19).217 Figure 17. Schematic representation for the synthesis of polymer-coated AuNPs by surface-initiated living-radical polymerization (LRP). Reprinted with permission from ref 214 (Fukuda���s group). Copyright 2002 American Chemical Society. Figure 18. SEM image of (a) 50-��m ceramic hollow spheres (CHSs), (b)-50 ��m gold-seeded CHSs, (c) 50-��m gold hollow spheres (GHSs) obtained by calcination and dissolution of gold-seeded CHSs, (d) a 100-��m CHS, (e) a 100-��m gold-seeded CHS, and (f) a 100-��m broken GHS. The arrow in (c) indicates a broken particle, which proves that it is hollow. From (f), it can be seen that the inside of GHS is empty. Reprinted with permission from ref 216b (Fendler���s group). Copyright 2002 Elsevier. Gold Nanoparticles Chemical Reviews, 2004, Vol. 104, No. 1 305

Nanosized domains of block copolymers can be used as nanoreactors to synthesize AuNPs by expansion of the nanosized domains and period of block copoly- mers, such as polystyrene-block-poly(4-vinylpyridine) (PS-PVP) diblock copolymers.218 Self-assemblies of AuNPs/polymer multilayer films have been formed using surface functionalization.219,220 AuNPs of aver- age size between 1 and 50 nm have also been stabilized by many water-soluble polymers, and some of then have been shown to be stable after 9 months in air. The most stable ones were obtained with polymers possessing hydrophobic backbones and side groups, allowing good interactions with the AuCl4- ion. The preparations were carried out using either UV irradiation or KBH4 to reduce HAuCl4 in the presence of a mass ratio of polymer:gold 25:1.221 Linear polymers having cyano or mercapto groups stabilize AuNPs of 1.5-3 nm diameter and narrow size distributions.222 AuNPs of Brust type with some thiol chain termini bearing exo-norbornene units were polymerized using ring-opening metathesis polymerization (ROMP) to produce a block copolymer shell.223,224 Small AuNPs (5 nm diameter) stabilized with sodium citrate225 were attached to the surface of silica nanoparticles protected by polymer layers to provide contrast in the final TEM image, a strategy also used to obtain TEM contrast for many types of molecular97 and biological materials.226 Solution be- havior, i.e., transformation in the morphology from small spherical AuNPs to large anisotropic objects, was observed by decreasing the concentration of polystyrene-block-poly(2-vinylpyridine) micelles be- low the critical micelle concentration (Figure 20).85 Networks of AuNPs prepared in water were ob- served by TEM upon adding poly(acrylic acid) to AuNPs stabilized by thiolated poly(ethylene oxide) chains of high molecular weight (necessary to stabi- lize AuNPs in water). Moreover, thin and linear thermally robust arrangements were formed when chondroitin sulfate c sodium salt (a polysaccharide carrying sulfuric acid groups and carboxylic acid groups) was added (Figure 21).227 AuNPs of about 20 nm size were formed upon reduction of AuCl3 by polyaniline in N-methylpyrrolidinone.228 An amine- functionalized polymer was used to simultaneously assemble carboxylic-acid-functionalized AuNPs and silica naoparticles into extended agregates.229 Such a strategy also led to spherical silica templates (Figure 22).226 Macroporous Au spheres with a di- ameter ���9 ��m have been formed by employing porous organic bead templates and preformed AuNPs.230 AuNPs were stabilized by the lone nitrogen pair on the backbone of polymethylphosphazene, [Me(Ph)- PN]n, and varying the ratio of [Me(Ph)PN]n to HAuCl4 Figure 19. AuNPs as templates for the synthesis of hollow polymer capsules. Reprinted with permission from ref 216c (Feldheim���s group). Copyright 1999 American Chemical Society. Figure 20. (a) Reaction scheme for the synthesis of the PEO-GMA-DEAR triblock copolymers. (b) Schematic illustration of the formation of three-layer ���onion-like��� micelles and shell cross-linked micelles from PEO-GMA-DEA triblock copolymers. PEO-GMA-DEA, poly[(ethylene oxide)-block-glycerol monomethacrylate-block-2-(diethylamino)ethyl methacrylate]. Re- printed with permission from ref 85 (Armes���s group). Copyright 2002 American Chemical Society. 306 Chemical Reviews, 2004, Vol. 104, No. 1 Daniel and Astruc

prior to reduction allowed control of the AuNP size.231 AuNPs (4-12 nm) were associated with thiol-func- tionalized polyoxometalates ��-[SiW10O36(RSi)O]4- (R ) HSC3H6), where the R group played the role of both stabilizing the AuNPs via the thiolate ligand and forming a covalent link to the polyanion through the trimethoxysilane group.232 The preparation of poly- (N-isopropylacrylamide)-protected AuNPs has been carried out in a homogeneous phase using various methods, and this polymer was found to be a better passivant than alkanethiols.233a AuNPs were prepared in both aqueous and organic systems by reducing HAuCl4 with o-anisidine in the presence of 1:1 N-methyl-2-pyrrolidone/toluene.233b AuNPs of 6 nm diameter and narrow size distribu- tions were stabilized by ��-conjugated poly(dithiaful- vene) polymers, and the oxidized form of this polymer induced a strong red shift of the absorption spectrum of the AuNPs to 550 nm (whereas the theory predicts 510-515 nm for the plasmon band in water).234 AuNPs with improved stability against long-term aggregation up to one month were prepared using poly(styrene)-block-poly(2-vinylpyridine) star-block copolymer.235 Water-soluble polymer-stabilized AuNPs were pre- pared from citrate-capped AuNPs by simple contact with dilute aqueous solutions of hydrophilic nonionic polymers based on the monomers N-[tris(hydroxy- methy)methyl]acrylamide and N-(isopropyl)acryl- amide that were functionalized with disulfide an- choring groups. The resulting polymer-coated AuNPs could be stored in the dry state and redispersed in water to yield sterically stabilized AuNP suspensions. The disulfide-bearing polymers exhibited only a slightly larger affinity for the gold surface than those that do not have the disulfide groups. The polymer layers allowed the free diffusion of small solutes but efficiently minimized the nonspecific absorption of large molecules such as proteins, a promising prop- erty (Figure 23).236a AuNPs have been synthesized in graft copolymer micelles,87 and the diffusion of AuNPs in a polymer matrix has been analyzed.236b Core-shell AuNPs have been prepared by the layer- by-layer technique, utilizing polyelectrolyte multi- layers assembled onto polystyrene cores as thin films in which to infiltrate AuNPs, and hollow spheres were obtained by removal of the templated polysty- rene cores.236c 3.12. Dendrimers A variety of assemblies between PAMAM dendrim- ers and AuNPs were reported in which the AuNPs were stabilized by the dendrimer that acted as both a polymer and a ligand. The AuNPs were stabilized only in the presence of excess PAMAM dendrimers and in solution, but PAMAM dendrimers function- alized with thiol termini could completely stabilize the AuNPs.237 PAMAM dendrimers were also func- tionalized with hydrophobic groups for solubilization of the AuNPs in organic solvents.238,368 Such den- drimer-AuNP assemblies were deposited as films on surfaces and used as sensors.239-241 AuNPs were synthesized from AuCl3 in DMF using PAMAM dendrimers that were modified with surface methyl ester groups.242 The use of PAMAM dendrimers for the stabilization of AuNPs allowed control of the Figure 21. TEM images of AuNPs covered with PEGSH 2000, observed in the presence of chondroitin sulfate c sodium salt (polysaccharide carrying sulfuric acid groups and carboxylic acid groups, which are expected to interact with the PEG chain). Reprinted with permission from ref 228 (Ishiwatari���s group). Copyright 2002 The Chemical Society of Japan. Figure 22. Schematic illustration for the synthesis of Au@HCMS (hollow core/mesoporous shell) polymer and carbon capsules. Reprinted with permission from ref 226 (Hyeon���s group). Copyright 2002 American Chemical So- ciety. Gold Nanoparticles Chemical Reviews, 2004, Vol. 104, No. 1 307