A Technique to Functionalize and Self-assemble Macroscopic Nanoparticle-ligand Monolayer Films onto Template-free Substrates

This protocol describes a self-assembly technique to create macroscopic monolayer films composed of ligand-coated nanoparticles1,2. The simple, robust and ...

Naval Research Laboratory

We examine the dependence of liquid crystalline nanocolloid formation and stability on surfactant. Nanocolloids composed of polymerizable liquid crystal mesogens and cross-linking agents and capped with either ionic or nonionic surfactants are prepared via the miniemulsion technique. Colloids synthesized with anionic surfactant were stable and displayed 2D hexagonal packing when deposited via slow vertical pulling of the silicon substrate from an aqueous suspension. Liquid crystal nanocolloids stabilized with the nonionic, polar polymer polyvinyl alcohol (PVA) were stable in aqueous environments but coalesced upon drying to form relatively large, well-defined crystal-like structures with uniform birefringence. SEM images reveal that the coalesced structures have mesalike features. Polarized light, atomic force, and polarized Raman microscopy of these structures indicate that the liquid crystal molecules are arranged with their long molecular axis slightly tilted with respect to the surface normal. A mechanism is proposed to explain the formation of the mesalike structures from the nanocolloids. These studies provide fundamental insight into the incorporation and stabilization of polymerizable liquid crystal molecules into nanovolumes and open up opportunities for the incorporation of functionality and anisotropy into isotropically shaped nanocolloids.

Role of surfactant in the stability of liquid crystal-based nanocolloids.

The controlled self-assembly of molecules and interactions between them remain a challenge in creating tunable and functional organic nanostructures. One class of molecular systems that has proven useful for incorporating tunable functionality at different length scales is liquid crystals (LCs) due to its ability to inherently self-organize. Here we present a novel approach to utilize the self-assembly of polymerizable liquid crystals to control the molecular aggregation of stable fluorescent chromophores and create a unique class of organic fluorescent nanocolloids. By adjusting the ratio between the dye and LC molecules inside the nanocolloids, we demonstrate the ability to control the molecular interactions and tune the fluorescent emission spectra of nanocolloid populations under single wavelength excitation. The single absorption spectrum and multiple emission spectra are highly desirable and reminiscent of the spectroscopic signature of quantum dots. These novel fluorescent nanocolloids have broad potential applications in fluorescent imaging and biological labeling.

Spectral tuning of organic nanocolloids by controlled molecular interactions.

We elucidate the polymer dynamics of a liquid crystal elastomer based on the time-dependent response of the pendent liquid crystal mesogens. The molecular tilt and switching time of mesogens are analyzed as a function of temperature and cross-linking density upon application of an electric field. We observe an unexpected maximum in the switching time of the liquid crystal mesogens at intermediate field strength. Analysis of the molecular tilt over multiple time regimes correlates the maximum response time with a transition to entangled polymer dynamics at a critical field strength.

Critical field strength in an electroclinic liquid crystal elastomer.

We present spectroscopic and optical studies of a non-layer-shrinkage ferroelectric liquid crystal DSiKN65. The orientational order parameters S, measured with respect to the smectic layer normal using IR spectroscopy on a sample aligned homeotropically, does not exhibit any significant variation between the smectic-A∗ and smectic-C∗ phases. In contrast the birefringence of a planar homogenous sample abruptly increases at the smectic-A∗ to smectic-C∗ transition. This suggests a general increase in the orientational order, which can be described by the orientational order parameters S' defined with respect to the director. Simultaneous increase of S' and the director tilt Θ may explain the low shrinkage of smectic layers, which is consistent with recent theoretical models describing the smectic-A∗ to smectic-C∗ transition for such materials.

Orientational order of a ferroelectric liquid crystal with small layer contraction.

We present a detailed analysis of the molecular packing of a strained liquid crystal elastomer composed of chiral mesogens in the smectic-A phase. X-ray diffraction patterns of the elastomer collected over a range of orientations with respect to the x-ray beam were used to reconstruct the three-dimensional scattering intensity as a function of tensile strain. We show that the smectic domain order is preserved in these strained elastomers. Changes in the intensity within a given scattering plane are due to reorientation, and not loss, of the molecular order in directions orthogonal to the applied strain. Incorporating the physical parameters of the elastomer, a nonlinear elastic model is presented to describe the rotation of the smectic-layered domains under strain, thus providing a fundamental analysis to the mechanical response of these unique materials.

Strain analysis of a chiral smectic-A elastomer.

Most natural and man-made fibers have circular cross-sections; thus the properties of materials composed of non-circular fibers are largely unexplored. We demonstrate the technology for fabricating fibers with predetermined cross-sectional shape. Passive hydrodynamic focusing and UV polymerization of a shaped acrylate stream produced metre-long fibers for structural and mechanical characterization.

UV polymerization of hydrodynamically shaped fibers.

We report the synthesis of colloidal nanoparticles with an internal structure forming a gel-like matrix. These nanoparticles are composed of low molecular weight liquid crystal (LC) 4-pentyl-4-cyanobiphenyl (5CB) encapsulated in an LC-based polymer network. Using nanoscopic mechanical analysis, we demonstrate the ability to independently tune the shape anisotropy and stiffness by varying, respectively, the 5CB concentration and the extent of the polymer cross-linking. Based on these data, a model is introduced to account for the effect of the polymer network on the mechanical properties, thus providing novel insight into the nanomechanics of these soft particles.

Tuning mechanical properties of liquid crystalline nanoparticles.

Barnacles adhere permanently to surfaces by secreting and curing a thin interfacial adhesive underwater. Here, we show that the acorn barnacle Balanus amphitrite adheres by a two-step fluid secretion process, both contributing to adhesion. We found that, as barnacles grow, the first barnacle cement secretion (BCS1) is released at the periphery of the expanding base plate. Subsequently, a second, autofluorescent fluid (BCS2) is released. We show that secretion of BCS2 into the interface results, on average, in a 2-fold increase in adhesive strength over adhesion by BCS1 alone. The two secretions are distinguishable both spatially and temporally, and differ in morphology, protein conformation, and chemical functionality. The short time window for BCS2 secretion relative to the overall area increase demonstrates that it has a disproportionate, surprisingly powerful, impact on adhesion. The dramatic change in adhesion occurs without measurable changes in interface thickness and total protein content. A fracture mechanics analysis suggests the interfacial material's modulus or work of adhesion, or both, were substantially increased after BCS2 secretion. Addition of BCS2 into the interface generates highly networked amyloid-like fibrils and enhanced phenolic content. Both intertwined fibers and phenolic chemistries may contribute to mechanical stability of the interface through physically or chemically anchoring interface proteins to the substrate and intermolecular interactions. Our experiments point to the need to reexamine the role of phenolic components in barnacle adhesion, long discounted despite their prevalence in structural membranes of arthropods and crustaceans, as they may contribute to chemical processes that strengthen adhesion through intermolecular cross-linking.

Barnacle Balanus amphitrite adheres by a stepwise cementing process.

Cell penetrating peptides facilitate efficient intracellular uptake of diverse materials ranging from small contrast agents to larger proteins and nanoparticles. However, a significant impediment remains in the subsequent compartmentalization/endosomal sequestration of most of these cargoes. Previous functional screening suggested that a modular peptide originally designed to deliver palmitoyl-protein thioesterase inhibitors to neurons could mediate endosomal escape in cultured cells. Here, we detail properties relevant to this peptide's ability to mediate cytosolic delivery of quantum dots (QDs) to a wide range of cell-types, brain tissue culture and a developing chick embryo in a remarkably nontoxic manner. The peptide further facilitated efficient endosomal escape of large proteins, dendrimers and other nanoparticle materials. We undertook an iterative structure-activity relationship analysis of the peptide by discretely modifying key components including length, charge, fatty acid content and their order using a comparative, semiquantitative assay. This approach allowed us to define the key motifs required for endosomal escape, to select more efficient escape sequences, along with unexpectedly identifying a sequence modified by one methylene group that specifically targeted QDs to cellular membranes. We interpret our results within a model of peptide function and highlight implications for in vivo labeling and nanoparticle-mediated drug delivery by using different peptides to co-deliver cargoes to cells and engage in multifunctional labeling.

Selecting improved peptidyl motifs for cytosolic delivery of disparate protein and nanoparticle materials.

Assembling DNA-based photonic wires around semiconductor quantum dots (QDs) creates optically active hybrid architectures that exploit the unique properties of both components. DNA hybridization allows positioning of multiple, carefully arranged fluorophores that can engage in sequential energy transfer steps while the QDs provide a superior energy harvesting antenna capacity that drives a Förster resonance energy transfer (FRET) cascade through the structures. Although the first generation of these composites demonstrated four-sequential energy transfer steps across a distance >150 Å, the exciton transfer efficiency reaching the final, terminal dye was estimated to be only ~0.7% with no concomitant sensitized emission observed. Had the terminal Cy7 dye utilized in that construct provided a sensitized emission, we estimate that this would have equated to an overall end-to-end ET efficiency of ≤ 0.1%. In this report, we demonstrate that overall energy flow through a second generation hybrid architecture can be significantly improved by reengineering four key aspects of the composite structure: (1) making the initial DNA modification chemistry smaller and more facile to implement, (2) optimizing donor-acceptor dye pairings, (3) varying donor-acceptor dye spacing as a function of the Förster distance R0, and (4) increasing the number of DNA wires displayed around each central QD donor. These cumulative changes lead to a 2 orders of magnitude improvement in the exciton transfer efficiency to the final terminal dye in comparison to the first-generation construct. The overall end-to-end efficiency through the optimized, five-fluorophore/four-step cascaded energy transfer system now approaches 10%. The results are analyzed using Förster theory with various sources of randomness accounted for by averaging over ensembles of modeled constructs. Fits to the spectra suggest near-ideal behavior when the photonic wires have two sequential acceptor dyes (Cy3 and Cy3.5) and exciton transfer efficiencies approaching 100% are seen when the dye spacings are 0.5 × R0. However, as additional dyes are included in each wire, strong nonidealities appear that are suspected to arise predominantly from the poor photophysical performance of the last two acceptor dyes (Cy5 and Cy5.5). The results are discussed in the context of improving exciton transfer efficiency along photonic wires and the contributions these architectures can make to understanding multistep FRET processes.

Achieving effective terminal exciton delivery in quantum dot antenna-sensitized multistep DNA photonic wires.

Photonic wires were constructed by sequentially arranging up to 7 fluorophores along a concatenated DNA scaffold. This yielded nanostructures displaying from one- to six-energy transfer steps where end-to-end efficiency reflected the multiple underlying photophysical processes and the ability of long-range interactions to compensate for localized non-ideal dye behaviour.

Christopher Spillmann

Naval Research Laboratory

A Technique to Functionalize and Self-assemble Macroscopic Nanoparticle-ligand Monolayer Films onto Template-free Substrates