A spatiotemporal characterization method for the dynamic cytoskeleton.

The significant gap between quantitative and qualitative understanding of cytoskeletal function is a pressing problem; microscopy and labeling techniques have improved qualitative investigations of localized cytoskeleton behavior, whereas quantitative analyses of whole cell cytoskeleton networks remain challenging. Here we present a method that accurately quantifies cytoskeleton dynamics. Our approach digitally subdivides cytoskeleton images using interrogation windows, within which box-counting is used to infer a fractal dimension (Df ) to characterize spatial arrangement, and gray value intensity (GVI) to determine actin density. A partitioning algorithm further obtains cytoskeleton characteristics from the perinuclear, cytosolic, and periphery cellular regions. We validated our measurement approach on Cytochalasin-treated cells using transgenically modified dermal fibroblast cells expressing fluorescent actin cytoskeletons. This method differentiates between normal and chemically disrupted actin networks, and quantifies rates of cytoskeletal degradation. Furthermore, GVI distributions were found to be inversely proportional to Df , having several biophysical implications for cytoskeleton formation/degradation. We additionally demonstrated detection sensitivity of differences in Df and GVI for cells seeded on substrates with varying degrees of stiffness, and coated with different attachment proteins. This general approach can be further implemented to gain insights on dynamic growth, disruption, and structure of the cytoskeleton (and other complex biological morphology) due to biological, chemical, or physical stimuli. © 2016 Wiley Periodicals, Inc.

Dissolution, agglomerate morphology, and stability limits of protein-coated silver nanoparticles.

Little is understood regarding the impact that molecular coatings have on nanoparticle dissolution kinetics and agglomerate formation in a dilute nanoparticle dispersion. Dissolution and agglomeration processes compete in removing isolated nanoparticles from the dispersion, making quantitative time-dependent measurements of the mechanisms of nanoparticle loss particularly challenging. In this article, we present in situ ultra-small-angle X-ray scattering (USAXS) results, simultaneously quantifying dissolution, agglomeration, and stability limits of silver nanoparticles (AgNPs) coated with bovine serum albumin (BSA) protein. When the BSA corona is disrupted, we find that the loss of silver from the nanoparticle core is well matched by a second-order kinetic rate reaction, arising from the oxidative dissolution of silver. Dissolution and agglomeration are quantified, and morphological transitions throughout the process are qualified. By probing the BSA-AgNP suspension around its stability limits, we provide insight into the destabilization mechanism by which individual particles rapidly dissolve as a whole rather than undergo slow dissolution from the aqueous interface inward, once the BSA layer is breached. Because USAXS rapidly measures over the entire nanometer to micrometer size range during the dissolution process, many insights are also gained into the stabilization of NPs by protein and its ability to protect the labile metal core from the solution environment by prohibiting the diffusion of reactive species. This approach can be extended to a wide variety of coating molecules and reactive metal nanoparticle systems to carefully survey their stability limits, revealing the likely mechanisms of coating breakdown and ensuing reactions.

Ultrafast, 2 min synthesis of monolayer-protected gold nanoclusters (d < 2 nm).

An ultrafast synthesis method is presented for hexanethiolate-coated gold nanoclusters (d < 2 nm, <250 atoms per nanocluster), which takes only 2 min and can be easily reproduced. With two immiscible solvents, gold nanoclusters are separated from the reaction byproducts fast and easily without any need for post-synthesis cleaning.

Challenges for physical characterization of silver nanoparticles under pristine and environmentally relevant conditions.

The reported size distribution of silver nanoparticles (AgNPs) is strongly affected by the underlying measurement method, agglomeration state, and dispersion conditions. A selection of AgNP materials with vendor-reported diameters ranging from 1 nm to 100 nm, various size distributions, and biocompatible capping agents including citrate, starch and polyvinylpyrrolidone were studied. AgNPs were diluted with either deionized water, moderately hard reconstituted water, or moderately hard reconstituted water containing natural organic matter. Rigorous physico-chemical characterization by consensus methods and protocols where available enables an understanding of how the underlying measurement method impacts the reported size measurements, which in turn provides a more complete understanding of the state (size, size distribution, agglomeration, etc.) of the AgNPs with respect to the dispersion conditions. An approach to developing routine screening is also presented.

Charged gold nanoparticles in non-polar solvents: 10-min synthesis and 2D self-assembly.

We report a fast and highly reproducible chemical synthesis method for colloidal gold nanoparticles which are negatively charged in nonpolar solvents and coated with hydrophobic organic molecules. If a hexane droplet containing charged gold nanoparticles is mixed with a larger toluene droplet, nanoparticles immediately float to the air-toluene interface and form a close-packed monolayer film. After evaporation of the solvent molecules, the monolayer film of nanoparticles can be deposited to any substrate without any limit in size. The synthesis does not require a postsynthesis cleaning step, since the two immiscible liquid phases separate the reaction byproducts from gold nanoparticles and a minimal amount of coating molecules is used.