Understanding Interactions between Cellular Matrices and Metal Complexes: Methods To Improve Silver Nanodot-Specific Staining.

Metal complexes are frequently used for biological applications due to their special photophysical and chemical characteristics. Due to strong interactions between metals and biomacromolecules, a random staining of cytoplasm or nucleoplasm by the complexes results in a low signal-to-background ratio. In this study, we used luminescent silver nanodots as a model to investigate the major driving force for non-specific staining in cellular matrices. Even though some silver nanodot emitters exhibited excellent specific staining of nucleoli, labeling with nanodots was problematic owing to severe non-specific staining. Binding between silver and sulfhydryl group of proteins appeared to be the major factor that enforced the silver staining. The oxidation of thiol groups in cells with hexacyanoferrate(III) dramatically weakened the silver-cell interaction and consequently significantly improved the efficiency of targeted staining.

Developing luminescent silver nanodots for biological applications.

Though creation and characterization of water soluble luminescent silver nanodots were achieved only in the past decade, a large variety of emitters in diverse scaffolds have been reported. Photophysical properties approach those of semiconductor quantum dots, but relatively small sizes are retained. Because of these properties, silver nanodots are finding ever-expanding roles as probes and biolabels. In this critical review we revisit the studies on silver nanodots in inert environments and in aqueous solutions. The recent advances detailing their chemical and physical properties of silver nanodots are highlighted with an effort to decipher the relations between their chemical/photophysical properties and their structures. The primary results about their biological applications are discussed here as well, especially relating to their chemical and photophysical behaviours in biological environments (216 references).

Open Linear Polymer Host-Guest Interactions Sensed by Luminescent Silver Nanodots.

The selectivity of the linear polymer chain toward its binding moieties has been considered negligible; thus, a clear demonstration showing the best-fit binding of a linear polymer to its guest counterpart is still unknown. Luminescent poly(acrylic acid) (PAA)-stabilized silver nanodots (PAA-AgNDs) have been applied as a turn-on sensor to monitor the interaction between the PAA chain and its binding cations. The binding of cations ions to the PAA chain may cross-link the linear PAA chain via coordination with carboxylate, which increases the rigidity of the polymer chain, retards the nonradiative decay of PAA-AgNDs, and consequently enhances the emission of silver nanodots while inducing a blue-shift of its emission spectrum. For the first time, we have demonstrated that a linear polymer chain can act as an open host to selectively bind to its best-matching cations. Specifically, among Group 2 cations (Mg2+, Ca2+, Sr2+, Ba2+), calcium ions show the strongest bonding to the PAA polymer chain. Our research suggests that, with extra rigidity, the polymer improves its chemical stability as calcium ions cross-linked the linear polymer. Meanwhile, it has also been demonstrated that luminescent silver nanodots can be excellent probes for the detection of polymer activities with straightforward and simple visualization methods.

Generation of luminescent noble metal nanodots in cell matrices.

Cell matrices were used as rich libraries to screen proteins for the production of luminescent silver and gold nanodots. The study indicates that the proteins for silver and gold nanodot protection are quite different. The identification of such proteins in future may enrich the family of luminescent nanodots.

Tuning the fast generation of luminescent silver nanodots on a surface.

Silver nanodots, predominantly a near-IR emitter, can be instantly generated on a surface by silver cluster transfer. Kinetic trapping of ssDNA molecules on the surface limits the reorganization of the resulting silver nanodots for other silver nanodot emitters. Adjusting the freedom of the adsorbed ssDNA can tune the generation of various silver nanodots on the surface.

Tailoring silver nanodots for intracellular staining.

Through tailored oligonucleotide scaffolds, Ag nanocluster syntheses have yielded thermally and cell-culture medium stable silver cluster-based emitters. Optimizing ssDNA stability has enabled creation of highly concentrated and spectrally pure nanocluster emitters with strong intracellular emission. Both fixed and live-cell staining become possible, and intracellular delivery is demonstrated both through conjugation to cell-penetrating peptides and via microinjection.

In Situ Generated Silver Nanodot Förster Resonance Energy Transfer Pair Reveals Nanocage Sizes.

Characterizing nanocages in macromolecules is one of the keys to understanding various biological activities and further utilizing nanocages for novel materials synthesis. However, fast and straightforward detection of the nanocage size remains challenging. Here, we present a new approach to detect the diameter of a nanocage by Förster resonance energy transfer (FRET) of luminescent silver nanodot pairs with reverse micelles as a model. Silver nanodot FRET pairs can be generated in situ from a single silver nanodot species with critical energy transfer distances, R0, of 4.8-6.5 nm. We have applied this approach to clarify the size variation of the water nanocage in nonionic surfactant Triton X-100-based reverse micelles. FRET efficiency decreases as more water is added, indicating that the size of the reverse micelles continuously expands with water content. The silver element in the nanocage also enhances the visualization of the nanocage under cryo-TEM imaging. The diameter of the water nanocage measured with the above approach is consistent with that obtained by cryo-TEM, demonstrating that the FRET measurement of silver nanodots can be a fast and accurate tool to detect nanocage dimensions. The above demonstration allows us to apply our strategy to other protein-based nanocages.

Hygroscopy-induced nanoparticle reshuffling in ionic-gold-residue-stabilized gold suprananoparticles.

Polyethyleneimine (PEI)-stabilized gold nanoparticles were used as a model to understand the roles of ionic precursors in the formation of nanoparticles and the impact of their presence on the nanoparticle properties. The low availability of elemental gold and the stabilization of the just-generated gold nanoparticles by the excess gold ions contributed to the production of ultra-small nearly neutral gold nanoparticles, resulting in properties significantly different from those prepared by conventional methods. The cross-linking between gold ions/PEI/nanoparticles further led to the assembly of these small gold nanoparticles into suprananoparticles that were stable in water. The hygroscopic Au(iii) residues in the suprananoparticles absorbed moisture to form a micro-water pool and the nanoparticles in the new aqueous solution reshuffled to generate larger nanoparticles, leading to significant changes in their optical properties. Such a phenomenon was formulated into a fast, sensitive and straightforward method for the detection of water content in organic solvents.

Oligonucleotide-stabilized Ag nanocluster fluorophores.

Single-stranded oligonucleotides stabilize highly fluorescent Ag nanoclusters, with emission colors tunable via DNA sequence. We utilized DNA microarrays to optimize these scaffold sequences for creating nearly spectrally pure Ag nanocluster fluorophores that are highly photostable and exhibit great buffer stability. Five different nanocluster emitters have been created with tunable emission from the blue to the near-IR and excellent photophysical properties. Ensemble and single molecule fluorescence studies show that oligonucleotide encapsulated Ag nanoclusters exhibit significantly greater photostability and higher emission rates than commonly used cyanine dyes.

Live cell surface labeling with fluorescent Ag nanocluster conjugates.

DNA-encapsulated silver clusters are readily conjugated to proteins and serve as alternatives to organic dyes and semiconductor quantum dots. Stable and bright on the bulk and single molecule levels, Ag nanocluster fluorescence is readily observed when staining live cell surfaces. Being significantly brighter and more photostable than organics and much smaller than quantum dots with a single point of attachment, these nanomaterials offer promising new approaches for bulk and single molecule biolabeling.

Silica nanoparticle stability in biological media revisited.

The stability of silica nanostructure in the core-silica shell nanomaterials is critical to understanding the activity of these nanomaterials since the exposure of core materials due to the poor stability of silica may cause misinterpretation of experiments, but unfortunately reports on the stability of silica have been inconsistent. Here, we show that luminescent silver nanodots (AgNDs) can be used to monitor the stability of silica nanostructures. Though relatively stable in water and phosphate buffered saline, silica nanoparticles are eroded by biological media, leading to the exposure of AgNDs from AgND@SiO2 nanoparticles and the quenching of nanodot luminescence. Our results reveal that a synergistic effect of organic compounds, particularly the amino groups, accelerates the erosion. Our work indicates that silica nanostructures are vulnerable to cellular medium and it may be possible to tune the release of drug molecules from silica-based drug delivery vehicles through controlled erosion.

Selective self-assembly of adenine-silver nanoparticles forms rings resembling the size of cells.

Self-assembly has played critical roles in the construction of functional nanomaterials. However, the structure of the macroscale multicomponent materials built by the self-assembly of nanoscale building blocks is hard to predict due to multiple intermolecular interactions of great complexity. Evaporation of solvents is usually an important approach to induce kinetically stable assemblies of building blocks with a large-scale specific arrangement. During such a deweting process, we tried to monitor the possible interactions between silver nanoparticles and nucleobases at a larger scale by epifluorescence microscopy, thanks to the doping of silver nanoparticles with luminescent silver nanodots. ssDNA oligomer-stabilized silver nanoparticles and adenine self-assemble to form ring-like compartments similar to the size of modern cells. However, the silver ions only dismantle the self-assembly of adenine. The rings are thermodynamically stable as the drying process only enrich the nanoparticles-nucleobase mixture to a concentration that activates the self-assembly. The permeable membrane-like edge of the ring is composed of adenine filaments glued together by silver nanoparticles. Interestingly, chemicals are partially confined and accumulated inside the ring, suggesting that this might be used as a microreactor to speed up chemical reactions during a dewetting process.

Ligand-assisted etching: the stability of silver nanoparticles and the generation of luminescent silver nanodots.

The etching mechanism of silver nanoparticles was investigated. Though a strong binding agent to silver and an oxidising agent synergistically etch the nanoparticles, the balanced etching power has to be optimized to generate luminescent silver nanodots. This indicates that active centres facilitate the formation of nanodots.

DNA-encapsulated silver nanodots as ratiometric luminescent probes for hypochlorite detection.

DNA-encapsulated silver nanodots are noteworthy candidates for bio-imaging probes, thanks to their excellent photophysical properties. The spectral shift of silver nanodot emitters from red to blue shows excellent correlations with the concentration of reactive oxygen species, which makes it possible to develop new types of probes for reactive oxygen species (ROS), such as hypochlorous acid (HOCl), given the outstanding stability of the blue in oxidizing environments. HOCl plays a role as a microbicide in immune systems but, on the other hand, is regarded as a disease contributor. Moreover, it is a common ingredient in household cleaners. There are still great demands to detect HOCl fluxes and their physiological pathways. We introduce a new ratiometric luminescence imaging method based on silver nanodots to sensitively detect hypochlorite. The factors that influence the accuracy of the detection are investigated. Its availability has also been demonstrated by detecting the active component in cleaners. PACS: 82; 82.30.Nr; 82.50.-m.

Oxidant-resistant imaging and ratiometric luminescence detection by selective oxidation of silver nanodots.

Reactive oxygen species selectively accelerated transitions between various silver nanodots. The blue was developed as an oxidant-resistant imaging agent and analyte reporter. In addition to the spectral response of nanodots to ROS, silver nanodots were formulated to detect analytes with excellent selectivity and picomolar detection limit when coupled to glucose oxidase.

Autofluorescence generation and elimination: a lesson from glutaraldehyde.

Glutaraldehyde causes especially high autofluorescence. It reacted with proteins and peptides to generate visible to near-IR emitters. A model indicated that ethylenediamine and a secondary amine in the molecule were key components for the formation of emissive species. The mechanism enables us to control the generation and elimination of autofluorescence.