Method for Improved Fluorescence Corrections for Molar Mass Characterization by Multiangle Light Scattering.

Multiangle light scattering (MALS) was used to determine the absolute molar mass of fluorescent macromolecules. It is standard protocol to install bandwidth filters before MALS detectors to suppress detection of fluorescent emissions. Fluorescence can introduce tremendous error in light scattering measurements and is a formidable challenge in accurately characterizing fluorescent macromolecules and particles. However, we show that for some systems, bandwidth filters alone are insufficient for blocking fluorescence in molar mass determinations. For these systems, we have devised a correction procedure to calculate the amount of fluorescence interference in the filtered signal. By determining the intensity of fluorescent emission not blocked by the bandwidth filters, we can correct the filtered signal accordingly and accurately determine the true molar mass. The transmission rates are calculated before MALS experimentation using emission data from standard fluorimetry techniques, allowing for the characterization of unknown samples. To validate the correction procedure, we synthesized fluorescent dye-conjugated proteins using an IR800CW (LI-COR) fluorophore and Bovine Serum Albumin protein. We successfully eliminated fluorescence interference in MALS measurements using this approach. This correction procedure has potential application toward more accurate molar mass characterizations of macromolecules with intrinsic fluorescence, such as lignins, fluorescent proteins, fluorescence-tagged proteins, and optically active nanoparticles.

Carbon Nanotube and Cellulose Nanocrystal Hybrid Films.

The use of cellulose nanocrystals (CNC) in high performance coatings is attractive for micro-scale structures or device fabrication due to the anisotropic geometry, however CNC are insulating materials. Carbon nanotubes (CNT) are also rod-shaped nanomaterials that display high mechanical strength and electrical conductivity. The hydrophobic regions of surface-modified CNC can interact with hydrophobic CNT and aid in association between the two anisotropic nanomaterials. The long-range electrostatic repulsion of CNC plays a role in forming a stable CNT and CNC mixture dispersion in water, which is integral to forming a uniform hybrid film. At concentrations favorable for film formation, the multiwalled nanotubes + CNC mixture dispersion shows cellular network formation, indicating local phase separation, while the single-walled nanotube + CNC mixture dispersion shows schlieren texture, indicating liquid crystal mixture formation. Conductive CNT + CNC hybrid films (5-20 µm thick) were cast on glass microscope slides with and without shear by blade coating. The CNT + CNC hybrid films electrical conductivity increased with increasing CNT loadings and some anisotropy was observed with the sheared hybrid films, although to a lesser extent than what was anticipated. Percolation models were applied to model the hybrid film conductivity and correlate with the hybrid film microstructure.

Phospholipid Bilayer Softening Due to Hydrophobic Gold Nanoparticle Inclusions.

Liposome-nanoparticle assemblies (LNAs) are vital in the context of novel targeted drug-delivery systems, in addition to investigating nanoparticle-lipid bilayer interactions. Quantifying membrane structural properties and dynamics in presence of nanoparticle inclusions provides a simple model to elucidate nanoparticle effects on membrane biophysical properties. We present experimental evidences of bilayer softening due to small hydrophobic gold nanoparticle inclusions. LNA structure has been investigated by a combination of cryo-transmission electron microscopy, dynamic light scattering, and small-angle neutron scattering. Neutron spin echo spectroscopy demonstrated a remarkable ∼15% bending modulus decrease for LNAs relative to pure liposomes. Clear dependence of bending modulus on gold nanoparticle diameter and concentration was observed from our observations. Our findings point toward local bilayer fluidization by nanoparticle inclusions leading to an overall bilayer softening. These findings add valuable information to liposomal drug-delivery vehicle design and membrane biophysics research.

Phase Behavior of Acetylated Cellulose Nanocrystals and Origins of the Cross-Hatch Birefringent Texture.

Cellulose nanocrystals hydrolyzed by hydrochloric acid and esterified by acetic acid produce acetylated cellulose nanocrystals (CNC-AA) with acetyl ester surface functional groups. While much attention has been paid to understanding the phase behavior (liquid crystal) of aqueous dispersions of sulfonated nanocrystals, relatively few studies have focused on CNC-AA dispersions. CNC-AA dispersions exhibit multiple phase regimes and markedly different phase behavior due to their lower surface charge. At concentrations above 5.0 × 10-4%vol, a decrease in the apparent diffusion coefficient indicates the onset of interparticle interactions and a transition from the dilute regime. From 0.003 to 0.31%vol, biphasic behavior is observed, consisting of a birefringent lower phase and disordered or isotropic upper phase. Small-angle neutron scattering was used to measure the growth of fractal structures with increasing concentration and indicates a two-dimensional assembly with short-range order in a plate-like assembled geometry. Above 0.31%vol, the dispersion transitions exhibit a cross-hatch birefringent texture, which is believed to exist as a fine-scale nematic that possesses frozen-in flow shear behavior, consistent with a glassy phase. This cross-hatch pattern is maintained in dried films, where atomic force microscopy and scanning electron microscopy reveal a layered sheet-like structure. Imaging also indicates that the basic unit of CNC-AA microstructure in the film consists of 0.5-1.5 µm scale aligned nanorod domains, which agrees with neutron scattering and the dimensions of each individual "hatch" in the birefringent texture observed by cross-polarized microscopy. The assembly of the nanorods into this layered structure and the fine-scale nematic birefringent cross-hatch texture is of significant fundamental interest, particularly since it differs greatly from cellulose nanocrystals with other surface chemistry and offers potential opportunities in other applications owing to the unique assembly.

Small Particle Driven Chain Disentanglements in Polymer Nanocomposites.

Using neutron spin-echo spectroscopy, x-ray photon correlation spectroscopy, and bulk rheology, we studied the effect of particle size on the single-chain dynamics, particle mobility, and bulk viscosity in athermal polyethylene oxide-gold nanoparticle composites. The results reveal a ≈25% increase in the reptation tube diameter with the addition of nanoparticles smaller than the entanglement mesh size (≈5 nm), at a volume fraction of 20%. The tube diameter remains unchanged in the composite with larger (20 nm) nanoparticles at the same loading. In both cases, the Rouse dynamics is insensitive to particle size. These results provide a direct experimental observation of particle-size-driven disentanglements that can cause non-Einstein-like viscosity trends often observed in polymer nanocomposites.

pH Triggered Recovery and Reuse of Thiolated Poly(acrylic acid) Functionalized Gold Nanoparticles with Applications in Colloidal Catalysis.

Thiolated poly(acrylic acid) (PAA-SH) functionalized gold nanoparticles were explored as a colloidal catalyst with potential application as a recoverable catalyst where the PAA provides pH-responsive dispersibility and phase transfer capability between aqueous and organic media. This system demonstrates complete nanoparticle recovery and redispersion over multiple reaction cycles without changes in nanoparticle morphology or reduction in conversion. The catalytic activity (rate constant) was reduced in subsequent reactions when recovery by aggregation was employed, despite unobservable changes in morphology or dispersibility. When colloidal catalyst recovery employed a pH induced phase transfer between two immiscible solvents, the catalytic activity of the recovered nanoparticles was unchanged over four cycles, maintaining the original rate constant and 100% conversion. The ability to recover and reuse colloidal catalysts by aggregation/redispersion and phase transfer methods that occur at low and high pH, respectively, could be used for different gold nanoparticle catalyzed reactions that occur at different pH conditions.

Reduced Reactivity of Amines against Nucleophilic Substitution via Reversible Reaction with Carbon Dioxide.

The reversible reaction of carbon dioxide (CO2) with primary amines to form alkyl-ammonium carbamates is demonstrated in this work to reduce amine reactivity against nucleophilic substitution reactions with benzophenone and phenyl isocyanate. The reversible formation of carbamates has been recently exploited for a number of unique applications including the formation of reversible ionic liquids and surfactants. For these applications, reduced reactivity of the carbamate is imperative, particularly for applications in reactions and separations. In this work, carbamate formation resulted in a 67% reduction in yield for urea synthesis and 55% reduction for imine synthesis. Furthermore, the amine reactivity can be recovered upon reversal of the carbamate reaction, demonstrating reversibility. The strong nucleophilic properties of amines often require protection/de-protection schemes during bi-functional coupling reactions. This typically requires three separate reaction steps to achieve a single transformation, which is the motivation behind Green Chemistry Principle #8: Reduce Derivatives. Based upon the reduced reactivity, there is potential to employ the reversible carbamate reaction as an alternative method for amine protection in the presence of competing reactions. For the context of this work, CO2 is envisioned as a green protecting agent to suppress formation of n-phenyl benzophenoneimine and various n-phenyl-n-alky ureas.

Quantitative measurement of ligand exchange on iron oxides via radiolabeled oleic acid.

Ligand exchange of hydrophilic molecules on the surface of hydrophobic iron oxide nanoparticles produced via thermal decomposition of chelated iron precursors is a common method for producing aqueous suspensions of particles for biomedical applications. Despite the wide use, relatively little is understood about the efficiency of ligand exchange on the surface of iron oxide nanoparticles and how much of the hydrophobic ligand is removed. To address this issue, we utilized a radiotracer technique to track the exchange of a radiolabeled (14)C-oleic acid ligand with hydrophilic ligands on the surface of magnetite nanoparticles. Iron oxide nanoparticles functionalized with (14)C-oleic acid were modified with poly(ethylene glycol) with terminal functional groups including, L-3,4-dihydroxyphenylalanine, a nitrated L-3,4-dihydroxyphenylalanine, carboxylic acid, a phosphonate, and an amine. Following ligand exchange, the nanoparticles and byproducts were analyzed using liquid scintillation counting and inductively coupled plasma mass spectroscopy. The labeled and unlabeled particles were further characterized by transmission electron microscopy and dynamic light scattering to determine particle size, hydrodynamic diameter, and zeta potential. The unlabeled particles were characterized via thermogravimetric analysis and vibrating sample magnetometry. Radioanalytical determination of the (14)C from (14)C-oleic acid was used to calculate the amount of oleic acid remaining on the surface of the particles after purification and ligand exchange. There was a significant loss of oleic acid on the surface of the particles after ligand exchange with amounts varying for the different functional binding groups on the poly(ethylene glycol). Nonetheless, all samples demonstrated some residual oleic acid associated with the particles. Quantification of the oleic acid remaining after ligand exchange reveals a binding hierarchy in which catechol derived anchor groups displace oleic acid on the surface of the nanoparticles better than the phosphonate, followed by the amine and carboxylic acid groups. Furthermore, the results show that these ligand exchange reactions do not necessarily occur to completion as is often assumed, thus leaving a residual amount of oleic acid on the surface of the particles. A thorough analysis of ligand exchange is required to develop nanoparticles that are suitable for their desired application.

Correlating Quantitative Measurements of Radical Production by Photocatalytic TiO2 with Daphnia magna Toxicity.

Increased use of titanium dioxide (TiO2 ) nanoparticles (NPs) in domestic and industrial applications has increased the risk for adverse environmental outcomes based on an elevated likelihood of organism exposure. Anatase TiO2 NP exposure to ultraviolet A (UV-A) radiation in aquatic environments generates radical oxygen species (ROS), which may ultimately be responsible for increased organism toxicity. We have identified and measured the 2 most relevant ROS species, hydroxyl and superoxide radicals, and described that ROS can be modeled using the highly reactive hydroxyl radical to provide an upper bound for toxicity. The TiO2 NPs were co-exposed to increasing natural organic matter (NOM) amounts (measured as concentration of dissolved organic carbon [DOC]) and simulated-sunlight UV-A intensities. Radical production rate was determined using fluorescence spectroscopy and was positively correlated with increases in TiO2 concentration and UV-A intensity, and negatively correlated with increased DOC concentration. Daphnia magna toxicity was also found to decrease with NOM addition, which is attributed to the decreased radical production rate with increased DOC concentrations. We demonstrate that the rate of ROS production from simulated-sunlight-irradiated TiO2 NPs can be quantified using relatively simple fluorescent techniques. We show that toxicity to TiO2 NP varies greatly with conditions, and that concentration alone is a poor predictor of toxicity. Describing toxicity/hydroxyl radical measurement may be a more accurate way to describe overall risk. We provide a framework for a simple model to describe toxicity/hydroxyl radical. These conclusions demonstrate the importance of considering exposure conditions as a means of risk management during TiO2 NP toxicity testing, waste management, and regulatory decisions. Environ Toxicol Chem 2021;40:1322-1334. © 2021 SETAC.

Effect of jet stretch and particle load on cellulose nanocrystal-alginate nanocomposite fibers.

Alginate fibers have found many applications such as the preparation of dressings to treat exuding wounds, drug delivery, enzyme immobilization, etc.; however, their use is limited due to poor mechanical properties. Cellulose nanocrystals (CNCs) were isolated from cotton and introduced into calcium alginate fibers with the goal of improving their strength and modulus. The isolated CNCs are elongated nanoparticles of crystalline cellulose with an average length of 130 nm with a standard deviation (s) of 63 nm, an average width of 20.4 nm (s = 7.8 nm), and an average height of 6.8 nm (s = 3.3 nm). The CNCs were mixed with an aqueous sodium alginate dope solution and wet spun into a CaCl(2) bath to form fibers. It was found that if the apparent jet stretch (ratio of the fiber draw velocity to extrusion velocity) is kept constant, addition of the nanocrystals reduces the tensile strength and modulus of the material; however, a small concentration of CNCs in the dope solution increases the tensile energy to break and enables an increase in the fiber spinning apparent jet stretch ratio by nearly 2-fold at up to 25% CNCs load; the maximum ratio of 4.6 is observed at 25 wt % CNC loading as compared to a maximum of 2.4 for the native alginate. Mechanical testing showed a 38% increase in tenacity and a 123% increase in tensile modulus with 10 wt % CNCs loading and an apparent jet stretch of 4.2. The data suggest that alignment of the nanocrystals in the composites is a key factor influencing the mechanical properties. CNCs have potential to become a biocompatible, renewable, and cost-effective solution to reinforce alginate fibers.

Effect of gold nanoparticle incorporation into oil-swollen surfactant lamellar membranes.

An oil-swollen surfactant membrane is employed to measure the effects of incorporated hydrophobically functionalized gold nanoparticles (AuNPs) on the structure and dynamics of the membranes. While maintaining an average AuNP diameter of approximately 5 nm, the membrane thickness was varied from 5 nm to 7.5 nm by changing the amount of oil in the membrane. The membranes become softer as the proportion of oil is increased, while the thickness fluctuations become slower. We attribute this to an increased fluctuation wavelength. Incorporation of AuNPs in the membrane induces membrane thinning and softening. Oil molecules surround the nanoparticles in the membrane and help their relatively homogeneous distribution. AuNPs significantly alter the membrane's structure and dynamics through thinning of the membrane, increased compressibility, and possible diffusion of AuNPs inside the membrane.

A spectroscopic and computational exploration of the cybotactic region of gas-expanded liquids: methanol and acetone.

Local compositions in supercritical and near-critial fluids may differ substantially from bulk compositions, and such differences have important effects on spectroscopic observations, phase equilibria, and chemical kinetics. Here, we compare such determinations around a solute probe dissolved in CO2-expanded methanol and acetone at 25 degrees C from solvatochromic experiments with molecular dynamics simulations. UV/vis and steady-state fluorescence measurements of the dye Coumarin 153 in the expanded liquid phase indicate preferential solvation in both the S0 and S1 states by the organic species. Simple dielectric continuum models are used to estimate local compositions from the spectroscopic data and are compared to molecular dynamics simulations of a single C153 molecule dissolved in the liquid phase at bubble point conditions. The simulations provide information about the local solvent structure around C153. They suggest the presence of large solvent clustering near the electron-withdrawing side of the probe. Preferential solvation exists in both the S0 and S1 states, but a large disagreement between simulation and experiment exists in the S1 state. Potential reasons for this disparity are discussed.

pH-Responsive Mercaptoundecanoic Acid Functionalized Gold Nanoparticles and Applications in Catalysis.

Mercaptoundecanoic acid (MUA) functionalized gold nanoparticles (AuNP-MUA) were synthesized and demonstrated to possess pH-triggered aggregation and re-dispersion, as well as the capability of phase transfer between aqueous and organic phases in response to changes in pH. The pH of aggregation for AuNP-MUA is consistent with the pKa of MUA (pH ~4) in solution, while AuNP-MUA phase transition between aqueous and organic phases occurs at pH ~9. The ion pair formation between the amine group in octadecylamine (ODA), the carboxylate group in MUA, and the hydrophobic alkyl chain of ODA facilitates the phase transfer of AuNP-MUA into an organic medium. The AuNP-MUA were investigated as a reusable catalyst in the catalytic reduction of 4-nitrophenol by borohydride-a model reaction for AuNPs. It was determined that 100% MUA surface coverage completely inhibits the catalytic activity of AuNPs. Decreasing the surface coverage was shown to increase catalytic activity, but this decrease also leads to decreased colloidal stability, recoverability, and reusability in subsequent reactions. At 60% MUA surface coverage, colloidal stability and catalytic activity were achieved, but the surface coverage was insufficient to enable redispersion following pH-induced recovery. A balance between AuNP colloidal stability, recoverability, and catalytic activity with reusability was achieved at 90% MUA surface coverage. The AuNP-MUA catalyst can also be recovered at different pH ranges depending on the recovery method employed. At pH ~4, protonation of the MUA results in reduced surface charge and aggregation. At pH ~9, ODA will form an ion-pair with the MUA and induce phase transfer into an immiscible organic phase. Both the pH-triggered aggregation/re-dispersion and aqueous/organic phase transfer methods were employed for catalyst recovery and reuse in subsequent reactions. The ability to recover and reuse the AuNP-MUA catalyst by two different methods and different pH regimes is significant, based on the fact that nanoparticle-catalyzed reactions may occur under different pH conditions.

Microelectromechanical Systems from Aligned Cellulose Nanocrystal Films.

Microelectromechanical systems (MEMS) have become a ubiquitous part of a multitude of industries including transportation, communication, medical, and consumer products. The majority of commercial MEMS devices are produced from silicon using energy-intensive and harsh chemical processing. We report that actuatable standard MEMS devices such as cantilever beam arrays, doubly clamped beams, residual strain testers, and mechanical strength testers can be produced via low-temperature fabrication of shear-aligned cellulose nanocrystal (CNC) films. The devices had feature sizes as small as 6 µm and anisotropic mechanical properties. For 4 µm thick doubly clamped beams with the CNC aligned parallel to the devices' long axes, the Young's moduli averaged 51 GPa and the fracture strength averaged 1.1 GPa. These mechanical properties are within one-third of typical values for polysilicon devices. This new paradigm of producing MEMS devices from CNC extracted from waste biomass provides the simplicity and tunability of fluid-phase processing while enabling anisotropic mechanical properties on the order of those obtained in standard silicon MEMS.

Solvent effects on AOT reverse micelles in liquid and compressed alkanes investigated by neutron spin-echo spectroscopy.

Neutron Spin-Echo (NSE) spectroscopy has been employed to study the interfacial properties of reverse micelles formed with the common surfactant sodium bis-2-ethylhexyl-sulfosuccinate (AOT) in liquid alkane solvents and compressed propane. NSE spectroscopy provides a means to measure small energy transfers for incident neutrons that correspond to thermal fluctuations on the nanosecond time scale and has been applied to the study of colloidal systems. NSE offers the unique ability to perform dynamic measurements of thermally induced shape fluctuation in the AOT surfactant monolayer. This study investigates the effects of the bulk solvent properties, water content, and the addition of octanol cosurfactant on the bending elasticity of AOT reverse micelles and the reverse micelle dynamics. By altering these solvent properties, specific trends in the bending elasticity constant, k, are observed where increasing k corresponds to an increase in micelle rigidity and a decrease in intermicellar exchange rate, k(ex). The observed corresponding trends in k and k(ex) are significant in relating the dynamics of microemulsions and their application as a reaction media. Compressed propane was also examined for the first time with a high-pressure, compressible bulk solvent where variations in temperature and pressure are used to tune the properties of the bulk phase. A decrease in the bending elasticity is observed for the d-propane/AOT/W = 8 reverse micelle system by simultaneously increasing the temperature and pressure, maintaining constant density. With isopycnic conditions, a constant translational diffusion of the reverse micelles through the bulk phase is observed, conforming to the Stokes-Einstein relationship.

Discrete nanoparticles induce loss of Legionella pneumophila biofilms from surfaces.

Nanoparticles (NPs) have been shown to induce dispersal events in microbial biofilms but the mechanism of the dispersal is unknown. Biofilms contaminate many man-made aquatic systems such as cooling towers, spas and dental lines. Within these biofilms, Legionella pneumophila is a primary pathogen, leading to these environments serving as sources for disease outbreaks. Here we show a reduction in biofilm bio-volume upon treatment with citrate-coated 6-nm platinum NPs, polyethylene glycol (PEG)-coated 11-nm gold NPs, and PEG-coated 8-nm iron oxide NPs. Treatment with citrate-coated 8-nm silver NPs, however, did not reduce biomass. The synthesis of NPs that remain dispersed and resist irreversible aggregation in the exposure media appears to be a key factor in the ability of NPs to induce biofilm dispersal.

Silver nanoparticle toxicity to Daphnia magna is a function of dissolved silver concentration.

The most persistent question regarding the toxicity of silver nanoparticles (AgNPs) is whether this toxicity is due to the nanoparticles themselves or the silver ions (Ag(+)) they release. The present study investigates the role of surface coating and the presence of dissolved organic carbon on the toxicity of AgNPs to Daphnia magna and tests the hypothesis that the acute toxicity of AgNPs is a function of dissolved Ag produced by nanoparticle dissolution. Toxicity of silver nitrate (AgNO3) and AgNPs with surface coatings-gum arabic (AgGA), polyethylene glycol (AgPEG), and polyvinylpyrrolidone (AgPVP)-at 48 h was assessed in US Environmental Protection Agency moderately hard reconstituted water alone and augmented with Suwannee River dissolved organic carbon (DOC). As expected, AgNO3 was the most toxic to D. magna and AgPVPs were the least toxic. In general, Suwannee River DOC presence reduced the toxicity of AgNO3, AgGAs, and AgPEG, while the toxicity of AgPVPs was unaffected. The measured dissolved Ag concentrations for all AgNPs and AgNO3 at the 48-h median lethal concentration in moderately hard reconstituted water were similar. The presence of Suwannee River DOC decreased the ratio of measured dissolved Ag to measured total Ag concentration. These results support the hypothesis that toxicity of AgNPs to D. magna is a function of dissolved Ag concentration from these particles.

Structural and thermal analysis of lipid vesicles encapsulating hydrophobic gold nanoparticles.

The structure and stability of hybrid lipid vesicles containing bilayer-encapsulated hydrophobic nanoparticles is dependent upon lipid phase behavior. By embedding stearylamine-stabilized gold nanoparticles in dipalmitoylphosphatidylcholine/dipalmitoylphosphatidylglycerol vesicles, we show that encapsulation at lipid to nanoparticle ratios from 10,000:1 to 5000:1 leads to bilayer thickening and hydrophobic mismatch, favoring nanoparticle inclusion in gel phase vesicles. High loadings lead to large increases in the gel to fluid melting temperature upon heating and significant hysteresis on cooling, which cannot be attributed solely to excess free ligand. This behavior is due to a cooperative effect of excess free SA ligand and nanoparticle embedment. Nanoparticle clustering was observed during lipid melting and could be reversed upon lipid freezing owing to lateral capillary forces within the bilayer. The impact of nanoparticle embedment on vesicle structure and properties at such low concentrations is reminiscent of hydrophobic proteins, suggesting that the underlying lipid biophysics between proteins and nanoparticle are similar and may provide a predictive design tool for therapeutic applications.

Green Synthesis of Robust, Biocompatible Silver Nanoparticles Using Garlic Extract.

This paper details a facile approach for the synthesis of stable and monodisperse silver nanoparticles performed at ambient/low temperature where Allium sativum (garlic) extract functions as the silver salt reducing agent during nanoparticle synthesis as well as the post-synthesis stabilizing ligands. Varying the synthesis conditions provides control of particle size, size-distribution, and kinetics of particle formation. Infrared spectroscopy, energy dispersive x-ray chemical analysis, and high performance liquid chromatography indicated that the carbohydrates present in the garlic extract are the most likely nanoparticle stabilizing chemistry. The synthesized silver nanoparticles also demonstrate potential for biomeical applications, owing to the 1) enhanced stability in biological media, 2) resistance to oxidation by the addition of H2O2, 3) ease and scalability of synthesis, and 4) lack of harsh chemicals required for synthesis. Cytotoxicity assays indicated no decrease in cellular proliferation for vascular smooth muscle cells and 3T3 fibroblasts at a concentration of 25 µg/ml, confirming that garlic extract prepared silver nanoparticles are ideal candidates for future experimentation and implementation into biomedical applications.

Probing the cybotactic region in gas-expanded liquids (GXLs).

Gas-expanded liquids (GXLs) are a new and benign class of liquid solvents, which may offer many advantages for separations, reactions, and advanced materials. GXLs are intermediate in properties between normal liquids and supercritical fluids, both in solvating power and in transport properties. Other advantages include benign nature, low operating pressures, and highly tunable properties by simple pressure variations. The chemical community has only just begun to exploit the advantages of these GXLs for industrial applications. This Account focuses on the synergism of experimental techniques with theoretical modeling resulting in a powerful combination for exploring chemical structure and transport in the cybotactic region of GXLs (at the nanometer lengthscale).