Dark-Field Microwells toward High-Throughput Direct miRNA Sensing with Gold Nanoparticles.

MicroRNA (miRNA) is a class of short RNA that is emerging as an ideal biomarker, as its expression level has been found to correlate with different types of diseases including diabetes and cancer. The detection of miRNA is highly beneficial for early diagnostics and disease monitoring. However, miRNA sensing remains difficult because of its small size and low expression levels. Common techniques such as quantitative real-time polymerase chain reaction (qRT-PCR), in situ hybridization and Northern blotting have been developed to quantify miRNA in a given sample. Nevertheless, these methods face common challenges in point-of-care practice as they either require complicated sample handling and expensive equipment, or suffer from low sensitivity. Here we present a new tool based on dark-field microwells to overcome these challenges in miRNA sensing. This miniaturized device enables the readout of a gold nanoparticle assay without the need of a dark-field microscope. We demonstrate the feasibility of the dark-field microwells to detect miRNA in both buffer solution and cell lysate. The dark-field microwells allow affordable miRNA sensing at a high throughput which make them a promising tool for point-of-care diagnostics.

Latchable microfluidic valve arrays based on shape memory polymer actuators.

We report arrays of latching microfluidic valves based on shape memory polymers (SMPs), and show their applications as reagent mixers and as peristaltic pumps. The valve design takes advantage of the SMP's multiple stable shapes and over a hundred-fold stiffness change with temperature to enable a) permanent zero-power latching in either open or closed positions (>15 h), as well as b) extended cyclic operation (>3000 cycles). The moving element in the valves consists of a tri-layer with a 50 µm thick central SMP layer, 25 µm thick patterned carbon-silicone (CB/PDMS) heaters underneath, and a 38 µm thick styrene ethylene butylene styrene (SEBS) impermeable film on top. Each valve of the array is individually addressable by synchronizing its integrated local Joule heating with a single external pressure supply. This architecture significantly reduces the device footprint and eliminates the need for multiplexing in microfluidic large scale integration (mLSI) systems.

Biocatalytically induced surface modification of the tobacco mosaic virus and the bacteriophage M13.

Engineered viruses are finding an increasing number of applications in basic, translational research and materials science. Genetic and chemical engineering of capsids represents a key point for tailoring the properties of viral particles, but the synthetic efforts and limits accompanying these processes still hinder their usability. Here, a single-step highly selective biocatalytic functionalization approach is described, providing a general platform for virus-acrylate hybrid particles. The tobacco mosaic virus (TMV) and the bacteriophage M13 have been successfully modified via laccase induced free radical formation on the tyrosine residues through single electron oxidation as the initiating step and the free radicals subsequently react with acrylate-based monomers. This new approach can be extended to other biomolecular assemblies with surface exposed tyrosine residues, when the introduction of new functionalities is desired.