Cryo-STEM-EDX spectroscopy for the characterisation of nanoparticles in cell culture media

We present a study of barium titanate nanoparticles dispersed in cell culture media. Scanning transmission electron microscopy combined with energy dispersive X-ray spectroscopy was undertaken on samples prepared using both conventional drop casting and also plunge freezing and examination under cryogenic conditions. This showed that drying artefacts occurred during conventional sample preparation, whereby some salt components of the cell culture media accumulated around the barium titanate nanoparticles; these were removed using the cryogenic route. Importantly, the formation of a calcium and phosphorus rich coating around the barium titanate nanoparticles was retained under cryo-conditions, highlighting that significant interactions do occur between nanomaterials and biological media

Examination of Combustion-Generated Smoke Particles from Biomass at Source: Relation to Atmospheric Light Absorption

The formation of carbonaceous aerosols from biomass combustion is associated with a high degree of uncertainty in global climate models. In this work, soot samples were generated from the combustion of pine wood, wheat straw and barley straw in a fixed bed stove; as well as from the combustion of biomass pyrolysis model compounds. Samples were collected on filters, which were used for the determination of Absorption Angstrom Exponent (AAE). In addition, the content and composition of elemental carbon (EC) and organic carbon (OC) were determined, and the interrelationships between these and the AAE were examined. It was found that the spectroscopic signature of samples with high ‘brown carbon’ emissions was comparable to that of many PAH and polyphenols, with AAE ranging from 1.0–1.2 for model compounds to 0.5–5.7 for biomass. In addition to the filter samples, particles were collected directly onto microscopy grids and analysed by transmission electron microscopy–electron energy loss spectroscopy (TEM-EELS) in order to determine structural characteristics. This was used to examine the impact of combustion conditions and flue gas dilution on particle structure. Smouldering phase and diluted particles were found to be less graphitic and twice as oxygenated as undiluted flaming phase particles. The results are interpreted to better understand the impact of combustion processes on soot formation from biomass combustion. Abbreviations: AAE: absorption angstrom exponent; ATN: light attenuation; AToFMS: aerosol time of flight mass spectrometer; BC: black carbon; BrC: brown carbon; C:O: carbon to oxygen ratio; CPD: cyclopentadienyl radical C5H5; DMS: differential mobility spectrometer; EC: elemental carbon; EELS: electron energy loss spectroscopy; HACA: hydrogen abstraction carbon addition; MCE: modified combustion efficiency; OA: organic aerosols; OC: organic carbon; PM: particulate matter; POM: primary (particulate)organic matter; Py-GC/MS: pyrolysis gas chromatography/mass spectrometry; sp2: amount of sp2 orbital hybridisation in atomic structure; SSA: single scattering albedo; TC: total carbon (BC+OC) or (EC+OC); TEM: transmission electron microscope; TGA: thermogravimetric analysis

Impact of Surface Ligand on the Biocompatibility of InP/ZnS Quantum Dots with Platelets

InP/ZnS quantum dots (QDs) have received a large focus in recent years as a safer alternative to heavy metal-based QDs. Given their intrinsic fluorescent imaging capabilities, these QDs can be potentially relevant for in vivo platelet imaging. The InP/ZnS QDs are synthesized and their biocompatibility investigated through the use of different phase transfer agents. Analysis of platelet function indicates that platelet-QD interaction can occur at all concentrations and for all QD permutations tested. However, as the QD concentration increases, platelet aggregation is induced by QDs alone independent of natural platelet agonists. This study helps to define a range of concentrations and coatings (thioglycolic acid and penicillamine) that are biocompatible with platelet function. With this information, the platelet-QD interaction can be identified using multiple methods. Fluorescent lifetime imaging microscopy (FLIM) and confocal studies have shown QDs localize on the surface of the platelet toward the center while showing evidence of energy transfer within the QD population. It is believed that these findings are an important stepping point for the development of fluorescent probes for platelet imaging

Understanding stress-induced disorder and breakage in organic crystals: beyond crystal structure anisotropy

Crystal engineering has advanced the strategies for design and synthesis of organic solids with the main focus being on customising the properties of the materials. Research in this area has a significant impact on large-scale manufacturing, as industrial processes may lead to the deterioration of such properties due to stress-induced transformations and breakage. In this work, we investigate the mechanical properties of structurally related labile multicomponent solids of carbamazepine (CBZ), namely the dihydrate (CBZ·2H2O), a cocrystal of CBZ with 1,4-benzoquinone (2CBZ·BZQ) and the solvates with formamide and 1,4-dioxane (CBZ·FORM and 2CBZ·DIOX, respectively). The effect of factors that are external (e.g. impact stressing) and/or internal (e.g. phase transformations and thermal motion) to the crystals are evaluated. In comparison to the other CBZ multicomponent crystal forms, CBZ·2H2O crystals tolerate less stress and are more susceptible to breakage. It is shown that this poor resistance to fracture may be a consequence of the packing of CBZ molecules and the orientation of the principal molecular axes in the structure relative to the cleavage plane. It is concluded, however, that the CBZ lattice alone is not accountable for the formation of cracks in the crystals of CBZ·2H2O. The strength and the temperature-dependence of electrostatic interactions, such as hydrogen bonds between CBZ and coformer, appear to influence the levels of stress to which the crystals are subjected that lead to fracture. Our findings show that the appropriate selection of coformer in multicomponent crystal forms, targetting superior mechanical properties, needs to account for the intrinsic stress generated by molecular vibrations and not solely by crystal anisotropy. Structural defects within the crystal lattice, although highly influenced by the crystallisation conditions and which are especially difficult to control in organic solids, may also affect breakage

Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™).

BACKGROUND: The rapid production and incorporation of engineered nanomaterials into consumer products alongside research suggesting nanomaterials can cause cell death and DNA damage (genotoxicity) makes in vitro assays desirable for nanosafety screening. However, conflicting outcomes are often observed when in vitro and in vivo study results are compared, suggesting more physiologically representative in vitro models are required to minimise reliance on animal testing. METHOD: BASF Levasil® silica nanoparticles (16 and 85 nm) were used to adapt the 3D reconstructed skin micronucleus (RSMN) assay for nanomaterials administered topically or into the growth medium. 3D dose-responses were compared to a 2D micronucleus assay using monocultured human B cells (TK6) after standardising dose between 2D / 3D assays by total nanoparticle mass to cell number. Cryogenic vitrification, scanning electron microscopy and dynamic light scattering techniques were applied to characterise in-medium and air-liquid interface exposures. Advanced transmission electron microscopy imaging modes (high angle annular dark field) and X-ray spectrometry were used to define nanoparticle penetration / cellular uptake in the intact 3D models and 2D monocultured cells. RESULTS: For all 2D exposures, significant (p < 0.002) increases in genotoxicity were observed (≥100 μg/mL) alongside cell viability decreases (p < 0.015) at doses ≥200 μg/mL (16 nm-SiO2) and ≥100 μg/mL (85 nm-SiO2). In contrast, 2D-equivalent exposures to the 3D models (≤300 μg/mL) caused no significant DNA damage or impact on cell viability. Further increasing dose to the 3D models led to probable air-liquid interface suffocation. Nanoparticle penetration / cell uptake analysis revealed no exposure to the live cells of the 3D model occurred due to the protective nature of the skin model's 3D cellular microarchitecture (topical exposures) and confounding barrier effects of the collagen cell attachment layer (in-medium exposures). 2D monocultured cells meanwhile showed extensive internalisation of both silica particles causing (geno)toxicity. CONCLUSIONS: The results establish the importance of tissue microarchitecture in defining nanomaterial exposure, and suggest 3D in vitro models could play a role in bridging the gap between in vitro and in vivo outcomes in nanotoxicology. Robust exposure characterisation and uptake assessment methods (as demonstrated) are essential to interpret nano(geno)toxicity studies successfully

Nanoparticle vesicle encoding for imaging and tracking cell populations.

For phenotypic behavior to be understood in the context of cell lineage and local environment, properties of individual cells must be measured relative to population-wide traits. However, the inability to accurately identify, track and measure thousands of single cells via high-throughput microscopy has impeded dynamic studies of cell populations. We demonstrate unique labeling of cells, driven by the heterogeneous random uptake of fluorescent nanoparticles of different emission colors. By sequentially exposing a cell population to different particles, we generated a large number of unique digital codes, which corresponded to the cell-specific number of nanoparticle-loaded vesicles and were visible within a given fluorescence channel. When three colors are used, the assay can self-generate over 17,000 individual codes identifiable using a typical fluorescence microscope. The color-codes provided immediate visualization of cell identity and allowed us to track human cells with a success rate of 78% across image frames separated by 8 h

Formation of stable uranium(VI) colloidal nanoparticles in conditions relevant to radioactive waste disposal

The favored pathway for disposal of higher activity radioactive wastes is via deep geological disposal. Many geological disposal facility designs include cement in their engineering design. Over the long term, interaction of groundwater with the cement and waste will form a plume of a hyperalkaline leachate (pH 10-13), and the behavior of radionuclides needs to be constrained under these extreme conditions to minimize the environmental hazard from the wastes. For uranium, a key component of many radioactive wastes, thermodynamic modeling predicts that, at high pH, U(VI) solubility will be very low (nM or lower) and controlled by equilibrium with solid phase alkali and alkaline-earth uranates. However, the formation of U(VI) colloids could potentially enhance the mobility of U(VI) under these conditions, and characterizing the potential for formation and medium-term stability of U(VI) colloids is important in underpinning our understanding of U behavior in waste disposal. Reflecting this, we applied conventional geochemical and microscopy techniques combined with synchrotron based in situ and ex situ X-ray techniques (small-angle X-ray scattering and X-ray adsorption spectroscopy (XAS)) to characterize colloidal U(VI) nanoparticles in a synthetic cement leachate (pH > 13) containing 4.2-252 μM U(VI). The results show that in cement leachates with 42 μM U(VI), colloids formed within hours and remained stable for several years. The colloids consisted of 1.5-1.8 nm nanoparticles with a proportion forming 20-60 nm aggregates. Using XAS and electron microscopy, we were able to determine that the colloidal nanoparticles had a clarkeite (sodium-uranate)-type crystallographic structure. The presented results have clear and hitherto unrecognized implications for the mobility of U(VI) in cementitious environments, in particular those associated with the geological disposal of nuclear waste

Nanoparticle modified polyacrylamide for enhanced oil recovery at harsh conditions

Silicon dioxide (SiO2) nanoparticles (NPs) have been recently proposed to increase the performance of polyacrylamide (PAM) for enhanced oil recovery (EOR) applications. However, SiO2/PAM nanocomposites tend to agglomerate or even desposit under harsh conditions such as high temperature-high salinity (HT-HS), which greatly decreases the potential for future field applications. In this work, SiO2 NPs were modified by (3-aminopropyl) triethoxysilane (M_SiO2) to create positively charged active groups that enabled strong interaction with PAM functional groups, leading to high dispersion stability. Three samples including M_SiO2/PAM, SiO2/PAM and NP-free PAM were synthesised in-situ via free radical polymerisation, and their thermal stability, rheological properties and the effect of aging time were studied. It was found that M_SiO2 could reduce the thermal degradation of the polymer and safeguard its backbone, resulting in much better thermal stability of PAM in harsh environments. After 90 days of aging, SiO2/PAM and NP-free PAM had 45 and 78% viscosity reduction; whereas only 10% reduction was observed for M_SiO2/PAM. In addition, core-flooding experiments showed that M_SiO2/PAM solutions produced more oil recovery than those from SiO2/PAM and NP-free PAM solutions at HT-HS condition