# Patent

## Title
Direct assembly of hydrophobic nanoparticles to multifunction structures

## Abstract

A process that allows convenient production of multifunctional composite particles by direct self-assembly of hydrophobic nanoparticles on host nanostructures containing high density surface thiol groups is present. Hydrophobic nanoparticles of various compositions and combinations can be directly assembled onto the host surface through the strong coordination interactions between metal cations and thiol groups. The resulting structures can be further conveniently overcoated with a layer of normal silica to stabilize the assemblies and render them highly dispersible in water for biomedical applications. As the entire fabrication process does not involve complicated surface modification procedures, the hydrophobic ligands on the nanoparticles are not disturbed significantly so that they retain their original properties such as highly efficient luminescence. Multifunctional nonspherical nanostructures can be produced by using mercapto-silica coated nano-objects of arbitrary shapes as hosts for immobilizing functional nanoparticles. Multilayer structures can be achieved by repeating the mercapto-silica coating and nanoparticle immobilization processes.

## Background
FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
This invention was made with government support under Grant No. DMR0956081 awarded by the National Science Foundation (NSF). The government has certain rights in this invention.


FIELD OF THE INVENTION
The invention relates to a method of producing multifunctional composite particles by direct self-assembly of hydrophobic nanoparticles on host nanostructures containing high density surface thiol groups, and the use of thiol-terminated colloidal organosilica (mercapto-silica) particles to directly immobilize hydrophobic ligand capped nanoparticles.
BACKGROUND
Multifunctional particles in the micrometer or sub-micrometer scale that exhibit two or more different properties are highly desirable for many important technological applications, ranging from catalysis to energy harvesting and transformation, multimodal imaging, detection, and simultaneous diagnosis and therapy. For example, microspheres embedded with magnetic iron oxide nanoparticles and fluorescent quantum dots have been widely studied as a multiple-mode imaging contrast agents combining magnetic resonance with optical detection and biological targeting. By carefully controlling the loading of quantum dots (QD), the obtained composite particles possess a dual function of optical encoding and magnetic separation. Replacing quantum dots with noble metal nanoparticles in such composites results in new types of multifunctional structures that are capable of magnetic resonance imaging and photothermal therapy. Magnetic materials have also been combined with nanocatalysts to form magnetically separable catalysts for the recovery and reuse of expensive catalysts after catalytic reactions, thus bridging the gap between heterogeneous and homogeneous catalysis.
Nanoparticle assembly represents a powerful approach that has been actively explored recently for producing bi-, tri-, and multifunctional materials in contrast to their limited single-component counterparts. By organizing different types of nanoparticles together, it not only allows the utilization of the size- and shape-dependent properties of individual nanoparticles, but also takes advantage of new properties resulting from the interactions between neighbors. Conventional assembly processes for multifunctional structures such as the well-known layer-by-layer (LbL) methods are mostly limited to hydrophilic nanoparticle systems because they heavily rely on electrostatic interactions. The direct linking nanoparticles through chemical bonds between surface ligands has also been attempted, but is limited to some special cases because most commonly used protecting ligands of nanoparticles do not contain additional active functional groups that allow further reactions.
Many technologically important high quality nanoparticles, especially semiconductors (such as CdSe, ZnSe, CdTe, and InP, InAs) and metal oxides (such as γ-Fe2O3, MnO, TiO2, ZrO2, CoFe2O4) are predominantly prepared through thermolytic routes by reacting inorganic precursors in organic solvents at high temperatures (preferably 150° C.-320° C.). The resulting nanostructures, however, retain the hydrophobic character of the organic ligands and, hence, are not soluble in water. As a result, it has been very difficult to assemble them directly into multifunctional nanostructures using means similar to those for water soluble particles. Typically, it is necessary to impose hydrophilic character on the nanoparticle surface and ensure water dispersibility (also bio-compatibility) by replacing the hydrophobic organic ligands with hydrophilic ones. However, ligand exchange processes usually involve several extra steps and in many cases are detrimental to the physical properties of the nanoparticles because the new hydrophilic ligands may not be able to effectively insulate the inorganic cores from the aqueous environment. For example, ligand exchange of hydrophobic trioctylphosphine oxide (TOPO) on the surface of CdSe/ZnS quantum dots (QDs) with various hydrophilic ligands causes a significant decrease in quantum efficiencies. Furthermore, the new ligands tend to desorb gradually from the nanoparticles, leading to aggregation and precipitation of the nanoparticles. More importantly, if more than one type of nanoparticles is needed for achieving multiple functions, each must be surface-treated separately before assembly. Therefore, it is highly desirable to develop a simple and general approach that allows the fabrication of multifunctional nanostructures by direct assembly of hydrophobic nanoparticles of various compositions. As their major applications are in the biomedical fields, it is also highly desired that these multifunctional systems be dispersible in water.
SUMMARY
In accordance with an exemplary embodiment, a method of production of multifunctional colloidal structures by direct self-assembly of nanoparticles on host nanostructures.
In accordance with another exemplary embodiment, use of thiol-terminated colloidal organosilica (mercapto-silica) particles to directly immobilize hydrophobic ligand capped nanoparticles.



BRIEF DESCRIPTION OF THE DRAWINGS
The accompanying drawings are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification. The drawings illustrate embodiments of the invention and, together with the description, serve to explain the principles of the invention. In the drawings,
FIG. 1 shows (a) schematic illustration showing the procedure of self-assembly of hydrophobic nanoparticles on MPS ((3-mercaptopropyl)trimethoxysilane spheres), (b) TEM images of oleic acid capped γ-Fe2O3 nanoparticles, and (c) MPS spheres, and (d) γ-Fe2O3 nanoparticles assembled on the surface of MPS spheres.
FIG. 2 shows (a) TEM images of various hydrophobic nanoparticles assembled on the surface of MPS spheres: left columns (column 1 and 3) show the different nanoparticles and right columns (column 2 and 4) represent the corresponding nanoparticle-capped MPS spheres, and wherein the ZrO2, TiO2 nanodots are originally protected by TOPO, NaYF4:Yb,Er, TiO2 nanorods by oleic acid, CdSe and NiO by TOPO and oleylamine, and both small and large Cu2S nanodisks by 1-dodecanethiol, and (b) SEM image and the corresponding EDX mapping of the elemental distribution of Si, Cd and Fe in a composite sphere of MPS@γ-Fe2O3&CdSe.
FIG. 3 shows the effect of original capping ligands on nanoparticle assembly, wherein (a) is the molecular structures of the three protection ligands capped on Au nanoparticles (TOAB, Cl2N and Cl2S), (b) digital photo of toluene dispersion of MPS spheres loaded with Au nanoparticles whose surfaces are capped with the three different protection ligands, and (c) TEM images of the corresponding MPS spheres in photo b.
FIG. 4 shows (a) a schematic diagram of the self-assembly of CdSe@CdZnS QDs on MPS spheres, showing how the PL spectra in (b) and TEM images in (c) were recorded, the original surface of QDs was covered by oleic acid, (b) PL spectra of the supernatant after adsorbtion of QDs by different amount of MPS spheres, and (c) TEM images of the precipitates (QD-capped MPS spheres) corresponding to the three points highlighted with numbers 1, 2 and 3 in FIG. b.
FIG. 5 shows (a) a schematic illustration showing the approach for transferring nanoparticle-capped MPS spheres from oil to water phase by forming bilayer structure and subsequent silica coating, (b) TEM images of low and high magnification of silica coated γ-Fe2O3 nanoparticle-capped MPS spheres (MPS@γ-Fe2O3@SiO2) synthesized in the presence of SDS, and wherein the γ-Fe2O3 nanoparticles were originally protected by oleic acid, and (c) TEM images of MPS spheres capped with various nanoparticles and then surface coated with silica in the presence of SDS surfactant: (1) MPS@Au@SiO2, (2) MPS@TiO2 nanorods@SiO2, and (3) MPS@ZrO2@SiO2, and wherein the Au nanoparticles were originally proteced by dodecylamine (Cl2N), TiO2 nanorods by oleic acid, and ZrO2 nanodots by TOPO.
FIG. 6 shows (a) TEM images of of γ-Fe2O3 nanoparticle-capped MPS spheres prepared in the presence of CTAB: (1) before silica coating; (2) after SiO2 coating with CTAB as the surfactant in aqueous NaOH solution; and (3) after additional SiO2 coating using the standard Stöber process, and wherein the γ-Fe2O3 nanoparticles were originally protected by oleic acid, and the inset in (2) is the corresponding TEM image at high magnification showing the presence of a thin silica layer (scale bar: 20 nm), (b) PL intensity as a function of time after storing the MPS@QD@SiO2 composite spheres in water under ambient conditions; wherein the inset is the TEM image of a portion of a MPS@QD@SiO2 sphere and the surface of CdSe@CdZnS QDs was originally covered by oleic acid, and (c) confocal optical microscopy image of 3.5-μm MPS spheres loaded with QDs of two different sizes, the QDs of different colors were assembled on MPS spheres separately and then mixed together for imaging, and wherein the sample was excited with a focused 488 nm line of Ar+ laser.
FIG. 7 shows (a,b) SEM image and EDX mapping (a) and TEM image (b) of the elemental distribution of Ag nanowire@SiO2&MPS@CdSe&γ-Fe2O3 composite nanowires, wherein the Ag nanowires were synthesized by using a polyol process in the presence of poly(vinyl pyrrolidone) (PVP), surface coated with normal silica using the Stöber process, modified with MPS layer, and then decorated with TOPO/oleylamine-capped CdSe and oleic acid-capped γ-Fe2O3 nanoparticles, and (c,d) SEM images and EDX mapping of the elemental distribution (c) and TEM image (d) of a MPS@γ-Fe2O3@SiO2&MPS@CdSe multilayer composite, synthesized by sequential procedures including initial loading of oleic acid capped γ-Fe2O3 nanoparticles on MPS spheres, surface coated with normal silica using the Stöber process, modified with MPS layer, and then decorated with TOPO/oleylamine capped CdSe nanoparticles, and wherein insets in (b) and (d) are schematic illustrations of the assembled structures.
FIG. 8 shows a schematic illustration showing the procedure of self-assembly of hydrophobic nanoparticles on MPS spheres in accordance with an exemplary embodiment.
FIG. 9 shows TEM images of γ-Fe2O3NPs assembled on the surface of MPS spheres, wherein the upper row shows the of γ-Fe2O3NPs with different size (5 nm, 12 nm and 20 nm) and the lower row represents the corresponding of γ-Fe2O3 NP capped MPS spheres.
FIG. 10 shows low magnification TEM image of ZrO2 nanoparticles assembled on the surface of MPS spheres.
FIG. 11 shows low magnification TEM image of CdSe and γ-Fe2O3 nanoparticles assembled on the surface of MPS spheres.
FIG. 12 shows low magnification TEM images of silica coated various composites assisted by SDS surfactant: (a) MPS@Au@SiO2, (b) MPS@TiO2 rods@SiO2 and (c) MPS@ZrO2@SiO2, and wherein the Au nanoparticles were originally proteced by dodecylamine (Cl2N), TiO2 nanorods by oleic acid, and ZrO2 nanodots by TOPO.
FIG. 13 shows FTIR spectra of normal silica spheres before and after coating with an additional MPS layer, and wherein the relatively strong absorption of C—H and S—H stretching indicates the high loading of thiol groups introduced during the modification.
FIG. 14 shows TEM images of (a) original Fe3O4@SiO2 particles and (b) corresponding Fe3O4@SiO2&MPS@CdSe@SiO2 composite structures, wherein the superparamagnetic Fe3O4 particles were synthesized using a high temperature precipitation reaction in the presence of polyacrylic acid (PAA), surface coated with normal silica using the Stöber process, modified with MPS layer, and then decorated with oleic acid-capped CdSe@CdZnS QDs, and then surface protected by another layer of normal SiO2 with the assistance of CTAB surfactant, and the insets are corresponding photographs of an aqueous dispersion of the composite particles excited by an ultraviolet lamp: the left sample is a normal dispersion while the right one is exposed to a magnetic field
FIG. 15 shows (a) a TEM image of silica coated Au and γ-Fe2O3 nanoparticles co-assembled composites (MPS@Au&γ-Fe2O3@SiO2), (b) UV-vis spectra showing gradual reduction of 4-NP by using MPS@Au&γ-Fe2O3@SiO2 composite catalyst, (c) plot of conversion of 4-nitrophenol as a function of reaction time in five successive cycles of reduction using magnetic recycling, showing that the silica protected Au nanoparticle catalysts can maintain their activity well in the five successive cycles, and wherein the conversion of 4-NP drops only slightly primarily because small portions of the solution are removed for sampling (approximately 3% each cycle).
FIG. 16 shows high and low magnification TEM images of Au (a) and Fe3O4 (b) hydrophilic nanoparticles assembly on the surface of MPS spheres, wherein the Au nanoparticles were originally covered by BSPP while the Fe3O4 nanoparticles were protected by PAA.



DETAILED DESCRIPTION
In accordance with an exemplary embodiment, a general process that allows convenient production of multifunctional composite particles by direct self-assembly of hydrophobic nanoparticles on host nanostructures containing high density surface thiol groups is described. Hydrophobic nanoparticles of various compositions and combinations can be directly assembled onto the host surface through the strong coordination interactions between metal cations and thiol groups.
In accordance with an exemplary embodiment, the resulting structures can be further conveniently overcoated with a layer of normal silica to stabilize the assemblies and render them highly dispersible in water for biomedical applications. As the entire fabrication process does not involve complicated surface modification procedures, the hydrophobic ligands on the nanoparticles are not disturbed significantly so that they retain their original properties such as highly efficient luminescence. Many complex composite nanostructures with tailored functions can be efficiently produced by using this versatile approach. For example, multifunctional nonspherical nanostructures can be efficiently produced by using mercapto-silica coated nano-objects of arbitrary shapes as hosts for immobilizing functional nanoparticles. Multilayer structures can also be achieved by repeating the mercapto-silica coating and nanoparticle immobilization processes. Such assembly approach will provide the research community a highly versatile, configurable, scalable, and reproducible process for the preparation of various multifunctional structures.
The strong coordinating interactions between alkanethiols and metal surfaces have been extensively studied and utilized in the formation of self-assembled monolayers (SAMs) on metal surfaces. It has also been reported that alkanethiols can adsorb onto metal oxide surfaces (such as TiO2, ZnO, and SnO2) and semiconductor substrates (such as InP, InAs, and GaAs, and metal sulfides). Recently, the thiol-metal interaction has been used to immobilize nanoparticles on alkanethiol-modified solid substrates, and has been further extended to the colloidal substrates by assembling Au nanoparticles to thiol-functionalized silica colloids. For example, Philipse et al. have explored the attachment of magnetic particles to colloidal silica that was grafted with (3-mercaptopropyl)trimethoxysilane (MPS). Their efforts, however, have been still limited to only water soluble γ-Fe2O3 and CoFe2O4 nanoparticles, probably due to the low density of thiol groups achievable during the surface modification of colloidal silica. In addition, prior studies have shown that colloidal silica with a high surface loading of thiol groups can effectively harvest hydrophobic Au nanocrystals from nonpolar solutions.
In accordance with an exemplary embodiment, the use of thiol-terminated colloidal organosilica (mercapto-silica) particles to directly immobilize various types of hydrophobic ligand capped nanoparticles, and further develop a general and powerful self-assembly method for the fabrication of multifunctional composite nanostructures is explored. Briefly, mercapto-silica colloidal particles made from MPS are first synthesized as adsorbent hosts, named as MPS spheres. Nanoparticles of desired functions are synthesized separately using well-established thermolysis methods, and then efficiently immobilized on the surface of MPS spheres by taking advantage of the high density thiol groups. In order to improve the chemical/mechanical stability and water-dispersibility, the composites can be further coated with a thin shell of normal silica. After demonstrating this simple assembly process, the procedure is generalized by immobilizing nanoparticles on mercapto-silica modified nanoscale objects, thus opening the door to the fabrication of more complex multifunctional composite structures.
FIG. 1a outlines the general self-assembly strategy for the fabrication of monodisperse, well-defined nanoparticle-silica multifunctional composites. The assembly involves two essential building blocks: mercapto-silica spheres and hydrophobic ligand-capped nanoparticles. In an exemplary process, a MPS precursor is hydrolyzed rapidly in an ammonia solution to yield spherical colloids. These mercapto-silica spheres are both structurally and functionally different from normal silica particles prepared from alkoxide precursors such as tetraethyl orthosilicate (TEOS). The use of MPS grants the resulting spheres a layer of high density surface thiol groups, which are expected to tether a dense layer of nanoparticles through the strong thiol-metal interactions. On the other hand, the abundant mercaptopropyl groups make the MPS spheres dispersible in nonpolar solvents such as toluene, facilitating the approaching of mercapto-silica and hydrophobic nanocrystals during the assembly process. The narrowly distributed MPS spheres with controlled sizes ranging from approximately 150 nm to approximately 3.5 micrometers can be produced by simply changing the reaction conditions, for example, the amount of the silane precursor, making it possible to produce uniform multifunctional nanocomposites with the desired dimensions. Hydrophobic ligand-capped nanoparticles are prepared through standard high-temperature thermolysis reaction. FIGS. 1b and 1c show transmission electron microscopy (TEM) images of oleic acid-capped γ-Fe2O3 nanoparticles (diameter: 12±0.7 nm) and MPS spheres (diameter: 280±30 nm).
Nanoparticles are assembled on the surface of MPS spheres by simply mixing them in nonpolar solvents such as toluene and cyclohexane. Generally, excess nanoparticles are used to ensure the dense coverage on the MPS spheres. Nanoparticle-capped MPS spheres are collected after washing several times with nonpolar solvent to remove free nanoparticles. As shown in FIG. 1d, a monolayer of γ-Fe2O3 nanoparticles can be clearly observed on the originally smooth surface of MPS spheres. No free nanoparticles can be observed, suggesting a strong attraction between the nanoparticles and MPS spheres. In accordance with an exemplary embodiment, during the assembly process, the thiol groups can partially replace the original surface ligands (oleic acid) and coordinate to the metal sites on the nanoparticle surface.
Thiols form stable bonds with most metal cations except alkali and alkaline earth ions. To test the versatility of the assembly process, various hydrophobic metal compound nanoparticles including trioctylphosphine oxide (TOPO) and oleylamine capped CdSe, TOPO capped ZrO2, oleic acid capped NaYF4:Yb,Er, TOPO and oleylamine capped NiO, 1-dodecanethiol capped Cu2S discs, TOPO capped TiO2 dots and oleic acid capped rods were assembled on the surface of the MPS spheres, as shown in FIG. 2a. A layer of nanoparticles can be easily identified on the surface of each MPS sphere, indicating the generality of the assembly strategy for hydrophobic metal compound nanoparticles. Note that Cu2S discs with different diameters (8 nm and 20 nm) show a similar high affinity for the MPS spheres, indicating a minimal size effect on the binding process. This has been also confirmed by the dense coverage of γ-Fe2O3 nanoparticles of 5 nm, 12 nm and 20 nm on MPS spheres. The similar adsorption behavior of TiO2 nanodots and nanorods on MPS spheres also suggests that the shape of nanoparticles has no significant effect on assembly efficiency.
The simple assembly process allows great flexibility in incorporating multiple nanoparticle components. A competition for immobilization on the surface of MPS spheres would appear among various types of nanoparticles due to their different affinity to thiol groups, which generally follows the Hard Soft Acid Base (HSAB) theory. Nanoparticles containing soft-acid cations such as Pt(II), Cu(I), Cd(II) and Au(I) bind to thiol groups much more strongly compared with those made of hard-acid cations such as Fe(III) and Ti(IV). In a nanoparticle mixture, it is necessary to increase the relative concentration of those with low-affinity to ensure sufficient loading. As an example shown in FIG. 2b, the assembly of γ-Fe2O3 nanoparticles and CdSe QDs on MPS spheres is demonstrated by mixing their dispersions at a particle number ratio of 10:1. The simultaneous capture of nanoparticles of γ-Fe2O3 and QD on the MPS surfaces has been confirmed by using scanning electron microscopy (SEM) imaging and energy dispersive X-ray (EDX) elemental mapping of a typical composite sphere. The strong Si signal across the sphere confirms the organosilica core, while the Fe and Cd signals both detected in the surface region clearly suggests the co-adsorption of these two types of particles.
In order to reach the metal sites on the nanoparticle surface, the thiol groups of MPS spheres may partially replace the original capping ligands on the nanoparticle surface. To evaluate this ligand replacement, hydrophobic Au nanoparticles capped with three different protection ligands, tetraoctylammonium bromide (TOAB), dodecylamine (Cl2N) and 1-dodecanethiol (Cl2S) are used for assembly. The molecular structures of these ligands are shown in FIG. 3a. After mixing with Au nanoparticles in toluene for approximately 20 minutes, the MPS spheres were separated from the mixture and washed a few times to remove free Au nanoparticles. FIG. 3b presents the digital photos of toluene dispersions of Au nanoparticle-immobilized MPS spheres, with TOAB, Cl2N, and Cl2S capped spheres exhibiting blue, purple, and light red colors, respectively. The shift of the surface plasmon resonance bands (color change) is the result of interparticle electromagnetic coupling: MPS spheres attract more TOAB capped Au nanoparticles, resulting in smaller interparticle distance and consequently a larger blue shift. The color intensity also indicates the loading of nanoparticles on MPS spheres. Due to the stability order of bonding Au—TOAB<Au—Cl2N<Au—Cl2S, the weaker capping ligands are detach easily from the Au surface to allow for the formation of Au—S bonds, leading to higher nanoparticle loading, as confirmed in FIG. 3c. The interesting fact that some Cl2S capped Au nanoparticles can still be captured by MPS spheres indicates that the thiol groups on MPS spheres can still replace some of the original alkanethiols to form new Au—S bonds even though the new bonds are similar to the original bonds in strength, which is consistent with the dynamic nature of the ligand adsorption on nanoparticle surface. As ligand replacement will eventually reach dynamic equilibrium, it can be predicted that nanoparticle immobilization can always occur even if the new thiol-nanoparticle bond is weak compared to the original ligand-nanoparticle interaction, but the actual loading may be significantly affected by the relative bond strength.
In accordance with an exemplary embodiment, the loading density of nanoparticles on the MPS spheres is related to their number ratio during assembly. If one assumes that the nanoparticles on the MPS spheres arrange in a hexagonal close-packing structure, the number of nanoparticles per MPS sphere, N, can be estimated by N=4α(1+R/r)2, where α=0.906 is the density of the circular hexagonal arrangement, and R and r are the radii of MPS spheres and nanoparticles, respectively. This equation allows one to estimate the amount of nanoparticles that is needed for a required loading density, for example, one MPS sphere with diameter of 300 nm can immobilize approximately 5×103 8-nm nanoparticles. Experimentally, one can quantify the nanoparticle loading by measuring their optical absorption or photoluminescence (PL), as shown in FIG. 4a by using CdSe/CdZnS QDs as an example. QDs approximately 8 nm in diameter in a toluene solution (500 μL, approximately 1014 particles per mL) were mixed with different amounts of MPS sphere solution (approximately 2×1010 spheres per mL) for 1 hour under vigorous shaking, followed by centrifugation at 11000 rpm for 10 min (free QDs remain in supernatant at this speed). Supernatant was collected for fluorescence measurement to monitor the amount of the free QDs left in the solution, and the precipitates were harvested for qualitative analysis of QD loading using TEM. As shown in FIG. 4b, the fluorescence intensity of the supernatant decreased as more MPS solution was added to the system. When only a small amount of MPS solution (<100 μL) was added, QDs remained in excess so that the decrease in fluorescence was nearly proportional to the added amount of MPS solution. The corresponding TEM image (FIG. 4c) confirms that the surface of every MPS sphere is fully occupied by QDs. Adding more MPS spheres (200 μL and 300 μL) led to a decrease in fluorescence intensity because more QDs were captured by MPS spheres and then removed from the supernatant. No free QDs were left in the supernatant when 300 μL of MPS spheres was added. At this stage there are not enough QDs to cover all the MPS surfaces in the form of a close-packed monolayer, as indicated in the TEM image in FIG. 4c. When 500 μL of MPS solution was added, even fewer QDs were adsorbed on each MPS sphere. If one assumes that 200 μL of MPS solution is the amount needed for a close-packed monolayer of QDs, each MPS sphere can capture 104 QDs as estimated from the concentration and volume of MPS spheres and QDs that are used. This value is consistent with the result calculated based on geometric considerations (5×103).
The adsorption of hydrophobic nanoparticles onto MPS spheres results in composite particles that are non-dispersible in water, thus preventing their direct application in environmental and biomedical applications. This limitation can be conveniently addressed by overcoating the composite particles with an additional layer of normal silica. Silica coating can endow the composites with biocompatibility and the possibility of subsequent functionalization. The additional advantages of using silica as a coating material mainly lie in its chemical stability, easy regulation of the coating process, controllable porosity, processability, and optical transparency.
The nanoparticle-capped MPS composites are first transferred to aqueous phase by using additional surfactants to adsorb onto the surface of composite spheres through the formation of bilayer structures, as illustrated in FIG. 5a. The phase transfer makes it convenient to subsequently grow a silica layer using the well-known Stöber method. The formation of a bilayer through hydrophobic-hydrophobic interaction also helps preserve the original ligands on the nanocrystal surface. For example, an anionic surfactant, sodium dodecyl sulfate (SDS), was used to mediate the silica coating of oleic acid capped MPS@γ-Fe2O3 composite spheres. After SDS adsorption, the MPS@γ-Fe2O3 composites were transferred into an ammonia/water/ethanol mixture and directly coated with uniform silica shells upon careful addition of TEOS. FIG. 5b displays typical images of SiO2 overcoated MPS@γ-Fe2O3 spheres. The thickness of the silica shell can be tuned from ten to several hundred nanometers by simply changing the amount of TEOS precursor. Interestingly, silica seems to initially nucleate on the individual nanoparticle surface and then grow independently before they eventually merge into a single layer. As a result, the silica coating possesses a rough surface until it reaches a thickness of approximately 100 nm. SDS has been demonstrated to be very effective in assisting the silica coating of composite spheres capped with various nanoparticles. This silica coating method assisted by SDS surfactant is universal for other hydrophobic nanoparticles-capped MPS spheres. In fact, the process has little to do with the compositions of nanoparticles and can be applied to passivate various nanoparticle/MPS systems as long as the additional surfactants can disperse these nanoparticles in the mixture of ethanol/H2O/NH3, which is the chemical environment required for the Stöber method for silica coating. FIG. 5c demonstrates a few more examples of normal silica passivated MPS@nanoparticle assemblies, with the nanoparticles being Au dots, TiO2 rods, and ZrO2 dots. Due to their small size compared to that of the MPS spheres, the nanoparticles cannot be individually identified in the TEM images and only a dark ring can be observed within the silica coating. Other exemplary surfactants, such as CTAB, can also be used to mediate silica coating on composite spheres. As shown in FIG. 6a with MPS@γ-Fe2O3 as an example, a previously reported method to first deposit a thin silica coating of a few nanometers on the composite surface by hydrolyzing TEOS in aqueous solution of CTAB with dilute NaOH solution as catalyst was adapted. The thickness of the silica layer can be further increased by an additional step of standard Stöber process. Unlike the case using SDS, this two-step procedure produces a very smooth silica surface even at a relatively small thickness. This procedure is especially useful for coating TOPO-capped MPS@QD spheres because CTAB causes smaller disturbance to the luminescence of QDs than SDS. FIG. 6b demonstrates the high stability of the PL of MPS@QD@SiO2 composites, with the TEM image of a portion of a typical sphere shown in the inset. The decrease of PL intensity is only about 10% even after 3 months of storage in water, which is significantly better than that of typical ligand-exchanged QDs. Surface passivation provided by silica and the intact ligand protection are the two main reasons for this excellent optical stability. FIG. 6c shows a multicolor fluorescence microscopy image of a mixture of two types of “monochromatic” MPS@QD@SiO2 spheres with an average diameter of 3.5 μm. These green and red spheres were prepared separately by using single-color quantum dots with emission wavelengths at 545 and 610 nm, respectively, and then mixed and imaged under a single wavelength light. All the spheres are observed with clearly distinguishable emission colors.
The necessity of MPS for high density thiol groups may appear as a limitation because MPS has only been produced in the form of microspheres and the functional nanoparticles can only be attached to the external surface of the spheres in the form of a monolayer. On the other hand, normal colloidal silica is well-known for its ability to form a conformal coating on various nanostructures with a high degree of control over the thickness. The assembly process comprises coating mercapto-silica onto the surface of other nanoscale objects so that further nanoparticle assembly can occur on hosts with more complex morphologies than spheres.
In accordance with an exemplary embodiment, the process includes first coating nano-objects such as dots, rods and wires with normal silica, and then introducing mercapto-silica to the surface through a co-condensation method. In the second step, multiple injections of the precursors are applied. The ratio of MPS to TEOS gradually increased for every injection to ensure a high density of thiol groups on the surface. The thiol-functionalized layer had been characterized by FTIR spectroscopy. After coating the normal silica spheres with an MPS layer, two bands in the ranges of 2950-2850 cm−1 and 2600-2500 cm−1 appeared, which can be attributed to the C—H and S—H stretching of the attached propanethiol groups. The relatively strong absorption of C—H and S—H stretching in the IR spectrum indicates that the loading of thiol groups is significantly higher than that of colloidal silica spheres modified with MPS silane using conventional grafting methods (where only a sub-monolayer of MPS can be attached).
The ability to form MPS coatings on normal silica surfaces opens the door to the fabrication of a large variety of multifunctional composite nanostructures. Functional materials can now be incorporated as cores inside normal silica, whose surface can be modified with high density thiol groups for attaching other types of functional nanoparticles. Here, a Fe3O4@SiO2&MPS@QD@SiO2 multilayer structure was demonstrated by using the simple solution phase assembly processes. First, superparamagnetic Fe3O4 particles with a diameter of approximately 100 nm were coated with SiO2. After coating with a MPS layer, QDs were assembled on the surface, and subsequently another layer of SiO2 was coated to make the composites water dispersible. The product exhibits strong fluorescence under UV excitation and superparamagnetic property that allows fast magnetic separation using an external magnet.
The process also allows the functionalization of nonspherical objects. As a demonstration, Ag@SiO2&MPS@γ-Fe2O3&QD multilayer composite nanowires were prepared using this simple assembly processes. Silver nanowires were synthesized using a polyol process, coated with normal silica, modified with MPS, and surface-immobilized with mixed nanoparticles of γ-Fe2O3 and QDs. FIG. 7a shows the elemental distribution of Si, O, Fe, Ag, Cd, and Se, as analyzed by EDX mapping. Ag@SiO2 cable-like structure can be easily identified by the strong Ag signal at the core and Si, O signals at the shell regions. The weaker but evenly distributed signals from Fe, Cd and Se prove the thin layer of immobilized γ-Fe2O3 and QDs. The uniform attachment of nanoparticles can be also clearly observed in the TEM image in FIG. 7b. The successful expansion to nonspherical morphologies makes this assembly process truly unique in comparison to conventional methods.
Analogous to the LbL approach, in accordance with an exemplary embodiment, the assembly process here also allows multilayer assembly by repeating the nanoparticle immobilization, silica coating, and MPS modification procedures. As a demonstration, MPS spheres (approximately 300 nm in diameter) were immobilized with γ-Fe2O3 nanoparticles on the surface, overcoated with a thin SiO2/MPS layer, and immobilized QDs on the surface again. FIGS. 7c and 7d show the EDX elemental mapping and a typical TEM image of a multilayer MPS@γ-Fe2O3@SiO2&MPS@QD structure. The dark ring marked with arrows is the layer of γ-Fe2O3 nanoparticles. The gray area outside this ring is the SiO2&MPS layer with thickness of approximately 50 nm. QDs can be clearly seen on the sphere surface. As indicated by the two dotted lines in FIG. 7c, the different locations of Fe and Cd (150 nm for Fe, 200 nm For Cd from the sphere's center) clearly suggests that the γ-Fe2O3 nanoparticles and QDs are distributed within different layers of the composite. The gap between these two nanoparticle layers is 50 nm which corresponds to the thickness of SiO2&MPS layer. Interestingly, a strong S signal is also observed in this gap, indicating high loading of thiol groups in this layer. All these observations confirm the complex structure with multiple types of nanoparticles positioned at different layers. This multilayer assembly strategy not only provides the means to improve the loading capacity, but also makes it convenient to increase the number of functions while maintaining the loading density of each component.
Materials and Methods
In accordance with exemplary embodiments, detailed synthesis recipes, additional TEM images of various MPS@NP composite spheres and silica passivated assemblies, FTIR spectra showing MPS coating, results demonstrating MPS@Au&γ-Fe2O3@SiO2 composite structures as magnetically recoverable catalysts, and TEM images showing the assembly of hydrophilic nanoparticles are described below.
Materials
Tetraethyl orthosilicate (TEOS), (3-mercaptopropyl)trimethoxysilane (MPS), sodium dodecyl sulfate (SDS), cetyl trimethylammonium bromide (CTAB), tetraoctylammonium bromide (TOAB), dodecylamine (Cl2N), 1-dodecanethiol (Cl2S), ammonium hydroxide solution (approximately 28% NH3 in water), oleylamine, trioctylphosphine oxide (TOPO), trioctylphosphine (TOP), tannic acid, sodium citrate tribasic dihydrate (99%), bis (p-sulfonatophenyl) phenylphosphine dihydrate dipotassium salt (BSPP, 97%), iron pentacarbonyl (Fe(CO)5), dioctyl ether, trifluoroacetic acid (TFA), phosphoric acid, cuprous acetate (CuOAc, 97%), 1-octadecene (ODE, 90%), Poly(acrylic acid) (Mw=1,800), cadmium nitrate tetrahydrate, sodium myristate, selenium, zinc oxide, cadmium oxide, sulfur, oleic acid and nickel(acetylacetonate)2 were purchased from Aldrich Chemical Co. Hydrogen tetrachloroaurate(III) trihydrate (HAuCl4.3H2O, 99.9+%) was purchased from Acros Organics. Tetrabutyl orthotitanate (TBOT) and titanium tetrachloride (TiCl4) were obtained from Fluka. Anhydrous iron (III) chloride (98%) was purchased from Riedel-de Haën. Diethylene glycol (reagent grade), ethyl alcohol (denatured), hydrochloric acid (36.5-38%), toluene (99.8%) and sodium hydroxide (98.8%), cyclohexane and acetone were obtained from Fisher Scientific. All chemicals were used as received without further treatment.
Synthesis of MPS Spheres
MPS spheres were prepared by using a one-pot procedure. (See Nakamura et al., J. Phys. Chem. C 2007, 111, 18892; and Lu et al., Langmuir 2011, 27, 3372.) In an exemplary synthesis, 120 μL MPS precursor was added to 30 mL of 1% ammonia aqueous solution and vigorously shaken for 1 minute at a speed of 2500 rpm. The mixture was then incubated at room temperature for 12 hours. The final products were washed with 0.01 M NaOH aqueous solution and ethanol several times and then dispersed in 10 mL of toluene. The size of MPS spheres can be tuned systematically by changing the amount of MPS precursor.
Synthesis of TiO2 Nanoparticles
TiO2 nanodots were prepared by a nonhydrolytic solution-based reaction. (See Trentler et al., J. Am. Chem. Soc. 1999, 121, 1613). Trioctylphosphine oxide (5 g) was heated at 150° C. for 5 minutes in vacuum. After increasing the solution temperature to 200° C. under N2 atmosphere, titanium tetrabutoxide (1.4 mL) was injected into the hot solution. The resulting mixture was then heated to 320° C., followed by rapidly adding 0.55 mL of TiCl4. The solution was further heated for 20 minutes at 320° C. to ensure the complete reaction. After cooling the system down to 80° C., 10 mL of acetone was added to yield a white precipitate, which was isolated by centrifugation and subsequently washed with a cylcohexane/acetone mixture to remove extra surfactant. The resulting powder was re-dispersed in 10 mL of cyclohexane.
Synthesis of TiO2 Nanorods
TiO2 nanorods were synthesized using a high-temperature pyrolysis reaction. (See Joo et al., J. Phys. Chem. B 2005, 109, 15297). Oleic acid (22 mL) was heated at 150° C. under vacuum for 1 hour. Then titanium isopropoxide (7 mL) was injected after the solution was cooled to room temperature under N2 flow. The mixture was then heated to 270° C. and kept at this temperature for 2 hours. Ethanol (40 mL) was added after the solution was cooled down to 80° C. The white precipitate was collected by centrifugation and washed with cyclohexane/ethanol mixture several times. The final product was re-dispersed in 20 mL of cyclohexane.
Synthesis of γ-Fe2O3 Nanocrystals
Superparamagnetic γ-Fe2O3 nanocrystals were synthesized using a thermolysis process. (See Hyeon et al., J. Am. Chem. Soc. 2001, 123, 12798). Fe(CO)5 (0.2 mL, 1.52 mmol) was added to a mixture containing 10 mL of octyl ether and 1.28 g of oleic acid at 100° C. The solution was then heated to 290° C. under an argon atmosphere and maintained at this temperature for 1 hour. After cooling down to 200° C., the solution was bubbled with air for 2 hours. After cooling down to room temperature, ethanol was added to the solution to precipitate γ-Fe2O3 nanoparticles, which were then separated by centrifugation. Finally, the resulting black powder was re-dispersed into 10 mL cyclohexane.
Synthesis of CdSe Nanoparticles
CdSe nanoparticles were prepared using a well established method reported by Peng group. (See Peng, et al., J. Am. Chem. Soc. 2002, 124, 3343). CdO (0.0254 g), stearic acid (0.228 g) and TOPO (3.88 g) were mixed in a 100 mL flask, then heated to 150° C. in vacuum for 30 min. After that, the mixture was heated to 320° C. under Ar flow to form light yellow solution. 2 mL oleylamine was injected later and after the temperature reached 320° C. again, the Se solution (0.722 g Se powder in 5 mL TOP) was quickly injected. And then, the reaction temperature was set at 290° C. and aged for 3 minutes. After the solution was quickly cooled down to 80° C., 5 mL of toluene was added. The CdSe nanoparticles were precipitated by ethanol and collected by centrifugation. After wash with toluene/ethanol one more time, the final product was re-dispersed into 5 mL toluene.
Synthesis of CdSe@CdZnS Nanoparticles (QD)
CdSe@CdZnS nanoparticles were prepared by a reproducible method reported by Dubertret group. (See Carion et al., Nat. Protocols 2007, 2, 2383). First, four reagents were prepared exactly based on the reference (1) Cadmium myristate (2) Cd(oleate)2 0.5 M in oleic acid (3) Zn(oleate)2 0.5 M in oleic acid and (4) TOPS 0.5 M solution in TOP. Later, 1-octadecene (16 mL), Cadmium myristate (170 mg) and selenium (12 mg) were mixed in a flask, and kept at room temperature in vacuum for 1 hour. After that, the solution was heated up to 240° C. under Ar atmosphere and aged for 10 min, followed by injection of 1 mL oleic acid and wait a further 2 minutes. After cooling the solution down to room temperature, the CdSe nanoparticles were precipitated by adding excess ethanol and then harvested by centrifugation. The resulting powder was re-dispersed in 10 mL of chloroform as a CdSe stock solution.
CdZnS shell coating: trioctylamine (8 mL), CdSe stock solution (2 mL), Zn(oleate)2 stock solution (400 μL) and Cd(oleate)2 stock solution (200 μL) were mixed in a flask. Then, the mixture was degassed under vacuum for 1 hour and heated to 300° C. under Ar atmosphere. Mixture of 4.4 mL trioctylamine and 0.6 mL TOPS was slowly added to the hot solution using a syringe pump at the rate of 10 mL/h. The final mixture solution was kept at 300° C. for 2 hours. The core/shell CdSe@CdZnS nanoparticles were precipitated by ethanol and finally dispersed in 5 mL of cyclohexane.

Synthesis of ZrO2 Nanoparticles

The preparation method is similar to the TiO2 nanopaticles. (See Joo et al., J. Am. Chem. Soc. 2003, 125, 6553). TOPO (10 g) was heated at 150° C. for 30 minutes in vacuum. After cool the solution temperature to 60° C. under N2 atmosphere, zirconium(IV) isopropoxide propanol complex (1.56 g) and ZrCl4 (1.16 g) were added into the solution. The resulting mixture was then heated to 340° C. and further heated for 2 hours at 340° C. to ensure the complete reaction. After cooling the system down to 80° C., 20 mL of acetone was added to yield a white precipitate, which was isolated by centrifugation and subsequently washed with a cylcohexane/acetone mixture to remove extra surfactant. The resulting powder was re-dispersed in 20 mL of cyclohexane.
Synthesis of NiO Nanoparticles
The NiO nanoparticles were synthesized by the hot injection method according to the previous report. (See Park et al., Adv. Mater. 2005, 17, 429). In an exemplary embodiment, TOP (5 g) was heated at 120° C. for 30 minutes in vacuum, and then increasing the temperature to 230° C. under Ar atmosphere. After that, a mixture of nickel(acetylacetonate)2 (0.5 g) and oleylamine (2 mL) was quickly injected and the solution was aged for 20 minutes. After cooling down to 160° C., the solution was bubbled with air for 1 hour. Ethanol was added to the solution to precipitate NiO nanoparticles, and then retrieved by centrifugation. The final black powder was redispersed in 10 mL cyclohexane after washing two times
Synthesis of NaYF4:Yb,Er Nanocrystals
Upconversion fluorescent NaYF4:18% Yb, 2% Er nanocrystals were synthesized according to the reference, Li et al., Adv. Mater. 2008, 20, 4765. YCl3.6H2O (242.7 mg), YbCl3.6H2O (70.0 mg) and ErCl3.6H2O (7.6 mg) were dissolved in 200 μL water first and then mixed with oleic acid (6 mL) and 1-octadecene (15 mL) in a 100 three-neck flask. The solution was degassed at 150° C. under Ar for 30 min, and then cooled down to room temperature. 10 mL of methanol solution containing NaOH (0.1 g) and NH4F (0.1481 g) was added and stirred for 30 minutes. After that, the solution was slowly heated to 110° C. and kept at 110° C. for 0.5 hours to remove methanol and small amount of water. During this period, one neck of the flask was left open under the flow of Ar gas. Then, the solution was quickly heated to 320° C. and aged for 1 hour under Ar protection. After the solution as cooled down, acetone was added to precipitate the nanoparticles. The final NaYF4:Yb,Er nanocrystals was redispersed in 5 mL of cyclohexane after washing with cyclohexane/acetone two times.
Synthesis of Cu2S nanoparticles
Cuprous acetate (0.0488 g), TOPO (1 g), and 1-octadecene (30 mL) were mixed in a three-neck flask and then degassed under argon flow for 30 min. Upon heating the solution up to 160° C., 1-dodecanethiol (1 mL) was injected quickly into the flask under vigorous stirring. The resulting mixture was further heated to 200° C. (for the small nanoparticles, the temperature is kept at 185° C.) and then reacted for 4 hours. The product was collected by adding an excess amount of methanol. After wash 2 to 3 times with toluene/methanol, the final Cu2S nanoparticles were dispersed in 5 mL toluene. (See Wang et al., Inorg. Chem. 2010, 49, 6601).
Synthesis of Hydrophobic Au Nanoparticles
Au nanoparticles capped with TOAB in toluene were synthesized by using biphasic reduction procedure. (See Brust et al., Chem. Commun. 1994, 801). In accordance with an exemplary synthesis, a biphasic TOAB solution was prepared by dissolving TOAB (0.365 g) in toluene (13.3 ml), followed by the addition of 5 mL HAuCl4 solution (0.01 g/mL). After vigorous stirring for 10 minutes, the color of the toluene phase became orange. After that, sodium borohydride solution (56.8 mg dissolved in 2 mL water) was added dropwise. The color changed from orange to colorless to purple to eventually reddish. The resulting mixture was further stirred for 2 hours to ensure monodispersity. The final products were washed with dilute H2SO4 (1M) one time and distilled water several times.
200 mg dodecylamine (Cl2N) or 1-dodecanethiol (Cl2S) was added to 2 mL of the above Au nanoparticle solution. The mixture was stirred for 3 to 4 hours at room temperature. Then 5 mL ethanol was added to precipitate the Au nanoparticles. After centrifugation, the Au nanoparticles can be redispersed in 2 mL toluene. In order to fully replace the original capping agent, this wash step can be repeated several times.
Assembly of Nanoparticles on MPS Spheres
Nanoparticles were assembled on the surface of MPS spheres by simply mixing them in nonpolar solvents such as toluene or cyclohexane. 500 μL of as-synthesized nanoparticles were added to 10 mL toluene dispersion of MPS spheres. Then, the mixture was votexed for 20 minutes at a speed of 500 rpm, followed by washing with nonpolar solvents a few times to remove free nanoparticles. Generally, the amount of nanoparticles is in excess compared to that of the MPS spheres.
For quantifying the QD loading on MPS spheres (FIG. 4), the concentration of as-synthesized QDs was calculated using the amount of limiting reagent selenium (2.4 mg), the density of CdSe, the final particle size and the volume of solvent toluene (5 mL). After diluted 100 times, the concentration of QDs is about 1014 nanoparticles per mL, which is consistent with the results measured from UV-Vis adsorption spectrum by Leatherdale et al. (See J. Phys. Chem. B 2002, 106, 7619). The concentration of the MPS spheres can be estimated at 2×1010 spheres per mL using the similar method. QD solution in toluene (500 μL) was mixed with different amount of MPS dispersion (0 to 500 μL) and pure toluene to obtain a total volume of 1 mL. After shaking 1 hour and subsequent centrifugation, PL of the supernatants was directly recorded and the precipitates were studied by TEM.
Silica Overcoating
For SDS assisted silica coating of γ-Fe2O3, Au, ZrO2 and TiO2 nanoparticle-capped MPS spheres, after washing and centrifugation, the precipitate was dissolved by 3 mL of aqueous solution of SDS (5.6 mg/mL) and sonicated for 10 minutes. The mixture was then heated to approximately 80° C. in a water bath for 1 hour to evaporate the remaining trace amount of toluene. The reaction solution was cooled down to room temperature and washed with water one time and re-dispersed in 3 mL of water. Then, ammonium hydroxide aqueous solution (1 mL, 28%), ethanol (20 mL), and TEOS (50 μL) were added to the solution sequentially. After stirring for 20 min, the core/shell colloids were collected by centrifugation and washed with ethanol three times.
For CTAB assisted silica coating, γ-Fe2O3 capped or QD-capped MPS spheres were first washed and centrifuged after assembly. Then 3 mL of aqueous solution of CTAB (10 mg/mL) was used to mediate the transfer of nanoparticle-capped MPS spheres to water. The composite spheres were re-dispersed in 15 mL H2O, mixed with aqueous NaOH solution (100 μL, 0.1 M), added 50 μL of methanol solution of TEOS (20% in volume) three times at 30-min intervals, reacted for 2 days under stirring, and then washed with ethanol three times. In order to grow a thicker silica layer, the thin-layer-silica coated composite particles were re-dispersed in water (3 mL), sequentially mixed with ammonia aqueous solution (1 mL, 28%), ethanol (20 mL), and TEOS (30 μL), and reacted for 20 min under vigorous stirring. The core/shell colloids were finally collected by centrifugation and washed with ethanol three times.
Coating of MPS Layer onto the Normal Silica Surface
Nanostructures covered with normal silica such as normal silica beads prepared by Stöber method, Fe3O4@SiO2, Ag wire@SiO2, or MPS@γ-Fe2O3@SiO2, with a preferred amount of approximately 20 mg, was dispersed in 3 mL water, mixed with 1 mL of ammonia (28%) and 20 mL of ethanol, after which a mixture of MPS and TEOS (20 μL, volume ratio 1:1) was added, and reacted for 20 min. After that, 20 μL of MPS/TEOS mixture (volume ratio: 5:1) was injected and reacted for another 20 minutes. The final products were washed with ethanol three times and re-dispersed in nonpolar solvent for subsequent nanoparticle immobilization.
Synthesis of Silica Coated Superparamagnetic Iron Oxide (Fe3O4@SiO2)
The Fe3O4 were synthesized in solution phase at high temperature. (See Ge et al., Angew. Chem. Int. Ed. 2007, 46, 4342). An NaOH/DEG stock solution was prepared by dissolving NaOH (50 mmol) in DEG (20 mL); this solution was heated at 120° C. for 1 hour under nitrogen, cooled, and kept at 70° C. A mixture of PAA (4 mmol), FeCl3 (0.4 mmol), and DEG (17 mL) was heated to 220° C. in a nitrogen atmosphere for 30 min with vigorous stirring to form a transparent, light yellow solution. NaOH/DEG stock solution (1.75 mL) was injected rapidly into the above hot mixture. The reaction solution was further heated at 220° C. for 1 hour. The final products were washed with a mixture of deionized water and ethanol several times and then dispersed in deionized water.
Fe3O4@SiO2 core-shell colloids were prepared through a modified Stöber process. (See Ge et al., Adv. Mater. 2008, 20, 3485). In an embodiment, an aqueous solution (3 mL) containing Fe3O4 (approximately 20 mg) was mixed with ethyl alcohol (20 mL), aqueous ammonia (28%, 1 mL) under vigorous magnetic stirring. TEOS (0.1 mL) was injected into the solution every 20 minutes until the total amount of TEOS reaches 0.5 mL.
Synthesis of Silica Coated Ag Wire (Ag wire@SiO2)
Silica coated Ag wire was prepared using a polyol process. (See Yin et al., Nano Lett. 2002, 2, 427). PtCl2 (2×10−5 g) was dissolved in 0.5 mL ethylene glycol and then added to 5 mL of ethylene glycol (heated to 160° C.) under continuous magnetic stirring. After 4 min, 2.5 mL ethylene glycol solution containing AgNO3 (0.05 g) and 5 mL ethylene glycol solution of PVP (MW approximately 40,000, 0.2 g) were added together to the ethylene glycol containing platinum seeds. This reaction mixture was then constantly heated at 160° C. for another 60 minutes. This mixture was centrifuged at approximately 4000 rpm to separate the silver nanowires from colloidal particles.
Approximately 2 mg of the silver nanowires were dispersed into a mixture of 20 mL 2-propanol, 4 mL deionized (DI) water and 0.4 of 28% ammonia solution. Then, 100 μL of TEOS was added to the mixture under magnetic stirring and reacted for 30 minutes. The silica coated silver wire can be collected by centrifugation at 4000 rpm and washed by ethanol for two times.
Preparation of Magnetic Catalyst (Calcined MPS@Au&γ-Fe2O3@SiO2)
500 μL of nanoparticles dispersion in cyclohexane (mixture of TOAB protected Au and γ-Fe2O3 nanoparticles at a particle number ratio of 10:1) were added to 10 mL toluene dispersion of MPS spheres. Then, the mixture was shaken for 20 min, followed by washing with nonpolar solvents a few times to remove free nanoparticles. After coating a layer of silica with assisted by SDS surfactant, the composites were dispersed in water and refluxed for 20 min to make the silica porous. (See Hu et al. Phys. Chem. Chem. Phys., 2010, 12, 11836). Later, the porous composites were calcined at 500° C. for 2 h to remove the protecting ligands on the nanoparticles.
Reduction of 4-nitrophenol Assisted by Calcined MPS@Au&γ-Fe2O3@SiO2 Catalyst
The reduction of 4-nitrophenol by NaBH4 was chosen as a model reaction for testing the efficiency of the prepared MPS@Au&γ-Fe2O3@SiO2 catalyst. Aqueous solutions of 4-nitrophenol (0.15 mL, 0.01M) and the calcined MPS@Au&γ-Fe2O3@SiO2 composite (5 mg) were added to de-ionized water (11 mL) in a vial under stirring. After adding aqueous NaBH4 (1 mL, 0.1 M) solution, the bright yellow solution gradually faded as the reaction progressed. UV-Vis spectra of the solution were recorded during the course of the reactions. A 1 mL sample was taken from the solution at 5, 10, 20, 30, 40 and 60 min for UVN is measurements to determine the conversion of 4-nitrophenol. The catalyst was quickly recovered from the solution using an external magnet field, rinsed with water and dispersed into water. The recovered catalyst was then re-dispersed into a mixture of new reactants to initiate another reaction cycle. The same procedure was performed in the additional 4 cycles. The performance and recyclability of the catalyst was shown in FIGS. 15b and 15c. When a small amount of the MPS@Au&γ-Fe2O3@SiO2 catalyst was introduced into the solution, the absorption at 400 nm decreased, indicating the reduction of 4-nitrophnol. As shown in FIG. 15c, the catalysts are still highly active after 5 successive cycles of the reaction. The multilayer nanocomposite structure containing both Au and γ-Fe2O3 nanoparticles designed here have several advantages as a catalyst. First, it can be recovered efficiently from the reaction solution by using external magnetic fields without significant losses. Second, the outer SiO2 framework can stabilize the catalyst nanoparticles and prevent their aggregations.
Synthesis of Hydrophilic Au Nanoparticles
Au nanoparticles were prepared by following published procedures. (See Liao et al., Appl. Phys. A: Mater. Sci. Process. 2003, 76, 541). Briefly, a 20 mL of aqueous solution containing 4 mL of 1% (w/w) trisodium citrate and 0.08 mL of 1% (w/w) tannic acid was rapidly added to an 80 mL aqueous solution containing 1 mL of 1% (w/w) hydrogen tetrachloroaurate(III) trihydrate at 60° C. Then the mixed solution was boiled for about 10 minutes under stirring. The solution was subsequently cooled down to room temperature (RT) with chilled water. A ligand exchange process was performed to enhance the stability of the as-synthesized Au nanoparticles. In brief, the AuNPs solution was mixed with an excess quantity of Bis(p-sulfonatophenyl) phenylphosphine dihydrate dipotassium (BSPP, with a concentration of 0.2 mg/mL), and then shaken overnight. (See Loweth, et al., Angew. Chem.-Int. Edit. 1999, 38, 1808). The resulting negatively charged Au nanoparticles are denoted as Au@BSPP.
Synthesis of Hydrophilic Fe3O4 Nanoparticles
Fe3O4 nanoparticles were prepared by mixing FeCl2.4H2O (0.4 g), FeCl3.6H2O (1.0 g) and 0.18 mL hydrochloric acid into 5 mL water and degassed with nitrogen before using. Then, the mixture was added to an aqueous NaOH solution (50 mL, 1.5 M) under the mechanical stir with N2 protection. After 0.5 h, the produced nanoparticles were centrifuged and rinsed with water several times, then re-dispersed in 20 mL water. CTAB was then coated to enhance the dispersity of the as-prepared Fe3O4 nanoparticles. The above dispersion of Fe3O4 nanoparticles (2 mL) was mixed with CTAB aqueous solution (10 mL, 10 mg/mL) and then sonicated for 0.5 to 1 hour. Then, the mixture solution was centrifuged at approximately 7000 rpm for 3 min twice for a size selection to discard the large Fe3O4 nanoparticles. The small Fe3O4 nanoparticles left in the supernatant were collected by an external magnet, rinsed with water three times and finally re-dispersed in 5 mL water. The pH value of the dispersion was tuned to approximately 4.0 by diluted HCl solution (0.001 M) for the later assembly on MPS spheres.
Assembly of Hydrophilic Nanoparticles on MPS Spheres
Hydrophilic nanoparticles can also be assembled on the surface of MPS spheres by a similar mixing method in polar solvents such as ethanol and water. MPS spheres were first washed with 0.01 M NaOH aqueous solution and subsequently deionized (D.I.) water two times and then dispersed in 2 mL of D.I. water. The final pH of the aqueous MPS dispersion is about 9 to 10. Au@BSPP nanoparticles were dispersed in a mixture of ethanol and water (v/v 9:1). In accordance with an exemplary embodiment, 50 μL of MPS dispersion was added to 1 mL of Au@BSPP or Fe3O4 nanoparticles solution. Then, the mixture was votexed for 20 minutes, followed by washing with water a few times to remove free Au@BSPP or Fe3O4 nanoparticles. The resulting TEM images of the hydrophilic nanoparticles assembly on MPS spheres are shown in FIG. 16.
In summary, a general process has been developed that allows convenient production of multifunctional colloidal particles by direct self-assembly of hydrophobic nanoparticles on host nanostructures containing mercapto-silica surfaces. The resulting composite structures can be further conveniently overcoated with a layer of normal silica to stabilize the assemblies and render them highly dispersible in water for biomedical applications. As the entire fabrication process does not involve complicated surface modification procedures, the hydrophobic ligands on the nanoparticles are not disturbed significantly so that they retain their original properties such as highly efficient luminescence. Many complex composite nanostructures with tailored functions can be conveniently produced by assembling nanoparticles of various compositions, sizes, and morphologies. Although the focus has been on the immobilization of hydrophobic nanoparticles as this has been a major challenge in the field, the developed process can be easily extended to the assembly of hydrophilic nanoparticles with minimal modifications. This universal, highly configurable, scalable and reproducible assembly process will help to produce various multifunctional structures for many important technological applications. Furthermore, nanoparticles of different compositions have different releasing behaviors depending on the strength of the specific nanoparticle-thiol interaction, thus promising for controlled drug delivery or related applications.
It will be understood that the foregoing description is of the preferred embodiments, and is, therefore, merely representative of the article and methods of manufacturing the same. It can be appreciated that many variations and modifications of the different embodiments in light of the above teachings will be readily apparent to those skilled in the art. Accordingly, the exemplary embodiments, as well as alternative embodiments, may be made without departing from the spirit and scope of the articles and methods as set forth in the attached claims.

## Claims

1. A method of production of multifunctional colloidal structures comprising:
assembling hydrophobic ligand capped nanoparticles on host nanostructures by direct self-assembly, the host nanostructures containing a mercapto-silica coating; and
applying the colloidal structures in a multilayer assembly by repeating the mercapto-silica coating and nanoparticle immobilization process on the host nanostructures, wherein different layers introduce different functionalization of the colloidal structures.

2. The method of claim 1, wherein a surface of the host nanostructures contain ligands that form coordination bonds with the hydrophobic ligand-capped nanoparticles.

3. The method of claim 1, wherein a surface of the host nanostructure contains thiol groups.

4. The method of claim 1, wherein a surface of the host nanostructure contains amine groups, or phosphine groups, or phosphate groups, or sulfate groups, or sulfonate groups, or carboxylate groups, or quaternary ammonium ions.

5. The method of claim 1, wherein the nanoparticles are hydrophobic.

6. The method of claim 1, wherein the nanoparticles are hydrophilic.

7. The method of claim 1, comprising:
overcoatinq the colloidal structures with a layer of silica to stabilize the assemblies of hydrophobic nanoparticles on the host nanostructures.

8. The method of claim 1, comprising:
functionalizing nonspherical particles by coating the nonspherical particles with a silica and modifying the silica coated nonspherical particles with a high-density thiol group.

9. The method of claim 1, comprising:
using thiol-terminated colloidal organosilica mercapto-silica particles to directly immobilize the hydrophobic ligand capped nanoparticles.

10. The method of claim 9, wherein the colloidal silica has a high-density surface loading of thiol groups.

11. The method of claim 1, wherein the mercapto-silica particles are mercapto-silica spheres.

12. The method of claim 11, comprising:
controlling a size of the mercapto-silica spheres from 150 nm to 3.5 micrometers.

13. The method of claim 1, comprising:
dispersing the colloidal structures in a nonpolar solvent.

14. The method of claim 1, wherein the mercapto-silica particles are formed by:
hydrolyzing a (3-mercaptopropyl)trimethoxysilane (MPS) precursor in an ammonia solution.

15. The method of claim 14, comprising:
immobilizing the mercapto-silica particles with Fe2O3 nanoparticles;
overcoating the mercapto-silica particles with the Fe2O3 nanoparticles with a layer of SiO2 and MPS; and
immobilizing the overcoated mercapto-silica particles with Fe2O3 nanoparticles with Fe2O3 nanoparticles.