Search for: All records

Fibrin is the main component of blood clots. The mechanical properties of fibrin are therefore of critical importance in successful hemostasis. One of the divalent cations released by platelets during hemostasis is Zn 2+ ; however, its effect on the network structure of fibrin gels and on the resultant mechanical properties remains poorly understood. Here, by combining mechanical measurements with three-dimensional confocal microscopy imaging, we show that Zn 2+ can tune the fibrin network structure and alter its mechanical properties. In the presence of Zn 2+ , fibrin protofibrils form large bundles that cause a coarsening of the fibrin network due to an increase in fiber diameter and reduction of the total fiber length. We further show that the protofibrils in these bundles are loosely coupled to one another, which results in a decrease of the elastic modulus with increasing Zn 2+ concentrations. We explore the elastic properties of these networks at both low and high stress: At low stress, the elasticity originates from pulling the thermal slack out of the network, and this is consistent with the thermal bending of the fibers. By contrast, at high stress, the elasticity exhibits a common master curve consistent with the stretching of individual protofibrils. These results show that the mechanics of a fibrin network are closely correlated with its microscopic structure and inform our understanding of the structure and physical mechanisms leading to defective or excessive clot stiffness. 

Although the utilization of rigid particles can afford stable emulsions, some applications require eventual emulsion destabilization to release contents captured in the particle-covered droplet. This destabilizing effect is achieved when using stabilizers that respond to controlled changes in environment. Microgels can be synthesized as stimuli responsive polymeric gel networks that adsorb to oil/water interfaces and stabilize emulsions. These particles are commonly hydrogels that swell and collapse in water in response to environmental changes. However, amphiphilic functionality is desired to enhance the adsorption abilities of these hydrogels while maintaining their stimuli responsivity. Microfluidic techniques are used to synthesize Janus microgels with two opposing stimuli responsive hemispheres. The particles have a temperature responsive domain connected to a pH responsive network where each side changes its hydrophilicity in response to a change in temperature or pH, respectively. The Janus microgels are amphiphilic in acidic conditions at 19 °C and alkaline conditions at 40 °C, while the opposite conditions cause a reduction of the amphiphilicity. By stabilizing emulsions with these dual responsive microgels, “smart” droplets that respond to environmental cues are formed. Emulsion droplets remain stable with smaller diameters when aqueous solution conditions favor amphiphilic particles yet, coalesce to larger droplets upon changing pH or temperature. These responsive Janus microgels represent the advancing technology of responsive droplets and demonstrate the applicability of microgels as emulsion stabilizers. 

Quantification of cell-secreted molecules, e.g. , cytokines, is fundamental to the characterization of immune responses. Cytokine capture assays that use engineered antibodies to anchor the secreted molecules to the secreting cells are widely used to characterize immune responses because they allow both sensitive identification and recovery of viable responding cells. However, if the cytokines diffuse away from the secreting cells, non-secreting cells will also be identified as responding cells. Here we encapsulate immune cells in microfluidic droplets and perform in-droplet cytokine capture assays to limit the diffusion of the secreted cytokines. We use microfluidic devices to rapidly encapsulate single natural killer NK-92 MI cells and their target K562 cells into microfluidic droplets. We perform in-droplet IFN-γ capture assays and demonstrate that NK-92 MI cells recognize target cells within droplets and become activated to secrete IFN-γ. Droplet encapsulation prevents diffusion of secreted products to neighboring cells and dramatically reduces both false positives and false negatives, relative to assays performed without droplets. In a sample containing 1% true positives, encapsulation reduces, from 94% to 2%, the number of true-positive cells appearing as negatives; in a sample containing 50% true positives, the number of non-stimulated cells appearing as positives is reduced from 98% to 1%. After cells are released from the droplets, secreted cytokine remains captured onto secreting immune cells, enabling FACS-isolation of populations highly enriched for activated effector immune cells. Droplet encapsulation can be used to reduce background and improve detection of any single-cell secretion assay. 

Droplet-based microfluidics is used to fabricate thin shell hydrogel microcapsules for the removal of methylene blue (MB) from aqueous solutions. The microcapsules composed of a poly(methacrylic acid) hydrogel shell exhibit unique properties, including permeation, separation, purification, and reaction of molecular species. Photocatalytic TiO 2 and ZnO nanoparticles encapsulated in the microcapsules, i.e. photocatalyst in capsule (PIC), are used to remove organic pollutants using an adsorption–oxidation mechanism. A prototype flow microreactor is assembled to demonstrate a controllable water purification approach in short time using photocatalysts. Our studies of aqueous and homogeneous hydrogel environments for the photocatalysts provide important insights into understanding the effectiveness of MB removal. Hydrogel capsules have MB removal rate comparable to homogeneous particles. Further reduction of both capsule and photocatalyst sizes can potentially aid in quicker water purification. 

Here, we demonstrate use of a Mg 2+ -dependent, site-specific DNA enzyme (DNAzyme) to cleave oligos from polyacrylamide gel beads, which is suitable for use in drop-based assays. We show that cleavage efficiency is improved by use of a tandem-repeat cleavage site. We further demonstrate that DNAzyme-released oligos function as primers in reverse transcription of cell-released mRNA.