Tissue Clearing To Localize Microplastics via Three-Dimensional Imaging of Whole Organisms.

Understanding the biological impacts of plastic pollution requires an effective methodology to detect unlabeled microplastics in environmental samples. Detecting unlabeled microplastics in an organism generally requires a digestion protocol, which results in the loss of spatial information on the distribution of microplastic within the organism and could lead to the disappearance of the smaller plastics. Fluorescence microscopy allows visualization of ingested microplastics but many labeling strategies are nonspecific and label biomass, thus limiting our ability to distinguish internalized plastics. While prelabeled plastics can be used to avoid nonspecific labeling, this approach precludes the detection of environmental microplastics in organisms. Also, using prelabeled microplastics can affect the viability of the organism and impact plastic uptake. Thus, a method was developed that employs nonspecific labeling with a tissue-clearing technique. Briefly, unlabeled microplastics are stained with a fluorescent dye after ingestion by the organism. The tissue-clearing technique then removes tissue-bound dye while rendering the structurally intact organism transparent. The internalized plastics remain stained and can be visualized in the cleared tissue with fluorescence microscopy. The technique is demonstrated using polystyrene beads in living aquatic organismsTigriopus californicusandDaphnia magnaand by spiking a model vertebrate (Cephalochordata) with different microplastics.

Effects of polydopamine-passivation on the optical properties of carbon dots and its potential use in vivo.

Passivation of carbon dots via heteroatom doping has been shown to enhance their optical properties and tune their fluorescence signature. Additionally, the incorporation of polymeric precursors in carbon dot synthesis has gained considerable interest with benefits to biological applications namely bioimaging, drug delivery and sensing, among others. In order to combine the desirable attributes of both, fluorescence enhancement and increased biocompatibility, polymers composed of high aromaticity and nitrogen content can be used as efficient carbon dot passivating agents. Here, the synthesis of fluorescent polymer-passivated carbon dots was developed through a microwave-assisted pyrolysis reaction of galactose, citric acid and polydopamine. Passivation of the dots with polydopamine induces a 90 nm red-shift in the fluorescence maxima from 420 to 510 nm. Moreover, passivation results in excitation-independent fluorescence and a 3.5-fold increase in fluorescence quantum yield, which increases from 1.3 to 4.6%. The application of the carbon dots as imaging probes was investigated in in vitro and in vivo model systems. Cytotoxicity studies in J774 and CHO-K1 cell lines revealed reduced cell toxicity for the polydopamine-passivated carbon dots in comparison to their unpassivated counterpart. In BALB/c mice, biodistribution studies demonstrated that regardless of surface passivation, the dots predominantly remained in the circulatory system 90 minutes post inoculation suggesting their potential use for cardiovascular therapies.

Fabrication of graphene-based porous materials: traditional and emerging approaches.

The anisotropic nature of 'graphenic' nanosheets enables them to form stable three-dimensional porous materials. The use of these porous structures has been explored in several applications including electronics and batteries, environmental remediation, energy storage, sensors, catalysis, tissue engineering, and many more. As method of fabrication greatly influences the final pore architecture, and chemical and mechanical characteristics and performance of these porous materials, it is essential to identify and address the correlation between property and function. In this review, we report detailed analyses of the different methods of fabricating porous graphene-based structures - with a focus on graphene oxide as the base material - and relate these with the resultant morphologies, mechanical responses, and common applications of use. We discuss the feasibility of the synthesis approaches and relate the GO concentrations used in each methodology against their corresponding pore sizes to identify the areas not explored to date.

Selective detection of nitrotyrosine using dual-fluorescent carbon dots.

The post-translational modification of amino acid plays a critical role in normal and diseased biological states. Specifically, nitrotyrosine (nTyr) has been linked to diseases, including neurodegeneration, among others. Hence, alternative methods are required for detection and differentiation of nTyr from other structurally similar analogues, such as Tyrosine (Tyr) or phosphotyrosine (pTyr). Herein, the selective detection of nTyr, over other congeners, was achieved by using dual-fluorescent carbon dots (CDs) in buffered solution, artificial saliva, bovine serum albumin and diluted equine serum. The nTyr induced fluorescence quenching of the blue and red emissions of CDs, in the 20-105 µM linear range, and with the limit of detection (LOD) at 34 µM, which was well below the physiological concentration required for detection. The sensor was functional at biological pH values, with optimal quenching efficiency at basic pH. The sensor was highly selective for nTyr even in the presence of common biological interferences (metal cations, organic anions, amino acids, nucleosides and other biologicals). The mechanism of quenching (a combination of static and dynamic) was ascribed to the nonradiative energy transfer, due to electronic overlap between nTyr absorbance and CDs fluorescence emission, and electron transfer from excited CDs state to nTyr as an electron acceptor. The dual-fluorescent CDs represent viable sensors for key biological modifications, and their selectivity and sensitivity may be further improved through tailored chemical synthesis of CDs, such as tunable surface chemistry to promote selective recognition of analyte of interest.

Elucidating the mechanism of dual-fluorescence in carbon dots.

Carbon dots have garnered significant attention owing to their versatile and highly tunable optical properties; however, the origins and the underlying mechanism remains a subject of debate especially for dual fluorescent systems. Here, we have prepared carbon dots from glutathione and formamide precursors via a one-pot solvothermal synthesis. Steady state and dynamic techniques indicate that these dual fluorescent dots possess distinct emissive carbon-core and a molecular states, which are responsible for the blue and red optical signatures, respectively. To further glean information into the fluorescence mechanism, electrochemical analysis was used to measure the bandgaps of the two fluorescent states, while femtosecond transient absorption spectroscopy evidenced the two-state model based on the observed heterogeneity and bimodal spectral distribution. Our findings provide novel and fundamental insights on the optical properties of dual fluorescent dots, which can translate to more effective and targeted application development particularly in bioimaging, multiplexed sensing and photocatalysis.

Insights into the Impact of Gold Nanoclusters Au10SG10 on Human Microglia.

The purpose of the current study is to uncover the impact of small liganded gold nanoclusters with 10 gold atoms and 10 glutathione ligands (Au10SG10) on several biomarkers in human microglia. We established the links connecting the atomically precise structure of Au10SG10 with their properties and changes in several biomolecules under oxidative stress. Au10SG10 caused the loss of mitochondrial metabolic activity, increased lipid peroxidation and translocation of an alarmin molecule, high mobility group box 1 (HMGB1), from the nucleus to the cytosol. Molecular modeling provided an insight into the location of amino acid interaction sites with Au10SG10 and the nature of bonds participating in these interactions. We show that Au10SG10 can bind directly to the defined sites of reduced, oxidized, and acetylated HMGB1. Further studies with similar complementary approaches merging live-cell analyses, determination of biomarkers, and cell functions could lead to optimized gold nanoclusters best suited for diagnostic and bioimaging purposes in neuroscience.

Green synthesis of carbon dots and their applications.

Carbon dots (CDs) are nanoparticles with tunable physicochemical and optical properties. Their resistance to photobleaching and relatively low toxicity render them attractive alternatives to fluorescent dyes and heavy metal-based quantum dots in the fields of bioimaging, sensing, catalysis, solar cells, and light-emitting diodes, among others. Moreover, they have garnered considerable attention as they lend themselves to green synthesis methods. Increasingly, one-pot syntheses comprising exclusively of renewable raw materials or renewable refined compounds are gaining favor over traditional approaches that rely on harsh chemicals and energy intensive conditions. The field of green CD synthesis is developing rapidly; however, challenges persist in ensuring the consistency of their properties (e.g., fluorescence quantum yield) relative to conventional preparation methods. This has mostly limited their use to sensing and bioimaging, leaving opportunities for development in optoelectronic applications. Herein, we discuss the most common green CD synthesis and purification methods reported in the literature and the renewable precursors used. The physical, chemical, and optical properties of the resulting green-synthesized CDs are critically reviewed, followed by a detailed description of their applications in sensing, bioimaging, biomedicine, inks, and catalysis. We conclude with an outlook on the future of green CD synthesis. Future research efforts should address the broad knowledge gap between CDs synthesized from renewable versus non-renewable precursors, focusing on discrepancies in their physical, chemical, and optical properties. The development of cost effective, safe, and sustainable green CDs with tunable properties will broaden their implementation in largely untapped applications, which include drug delivery, photovoltaics, catalysis, and more.

Intracellular ratiometric temperature sensing using fluorescent carbon dots.

Highly sensitive non-invasive temperature sensing is critical for studying fundamental biological processes and applications in medical diagnostics. Nanoscale-based thermometers are promising non-invasive probes for precise temperature sensing with subcellular resolution. However, many of these systems have limitations as they rely on fluorescence intensity changes, deconvolution of peaks, or the use of hybrid systems to measure thermal events. To address this, we developed a fluorescence-based ratiometric temperature sensing approach using carbon dots prepared via microwave synthesis. These dots possess dual fluorescence signatures in the blue and red regions of the spectrum. We observed a linear response as a function of temperature in the range of 5-60 °C with a thermal resolution of 0.048 K-1 and thermal sensitivity of 1.97% C-1. Temperature-dependent fluorescence was also observed in HeLa cancer cells over a range of 32-42 °C by monitoring changes in the red-to-blue fluorescence signatures. We demonstrate that the ratiometric approach is superior to intensity-based thermal sensing because it is independent of the intracellular concentration of the optical probe. These findings suggest that dual-emitting carbon dots can be an effective tool for in vitro and possibly in vivo fluorescence nanothermometry.

Facile Aqueous-Phase Synthesis of an Ultrasmall Bismuth Nanocatalyst for the Reduction of 4-Nitrophenol.

Bismuth metallic nanoparticles have evoked considerable interest in catalysis owing to their small size, high surface area-to-volume ratio, and low toxicity. However, the need for toxic reductants and organic solvents in their synthesis often limits their desirability for application development. Here, we describe a green strategy to synthesize bismuth nanodots via the redox reactions between bismuth nitrate and d-glucose, in the presence of poly(vinylpyrrolidone) in the basic aqueous phase. Both reagents play a crucial role in the formation of monodisperse bismuth nanodots acting as mild reducing and capping agents, respectively. We further demonstrate that the catalytic activity of these dots via the successful reduction of the environmental contaminant 4-nitrophenol to its useful 4-aminophenol analogue requiring only 36 µg/mL nanocatalyst for 20 mM of the substrate. Moreover, they can be recovered and recycled in multiple reactions before the onset of an appreciable loss of catalytic activity. The proposed facile synthetic route and inexpensive matrix materials lead the way to access bismuth nanodots for both the fundamental study of reactions and their industrial catalysis applications.