Quantification of colloidal filtration of polystyrene micro-particles on glass substrate using a microfluidic device.

A microfluidic device was designed to investigate filtration of particles in an electrolyte in the presence of liquid flow. Polystyrene spheres in potassium chloride solution at concentrations of 3-100 mM were allowed to settle and adhere to a glass substrate. A particle free solution at the same concentration was then flushed through the microfluidic channel at 0.03-4.0 mL/h. As the hydrodynamic drag on the adhered particles exceeded the intersurface interaction with the substrate, "pull-off" occurred and the particles detached. Filtration efficiency, α, was shown to a function of both ionic concentration of the liquid medium and flow speed, consistent with a phenomenological model based on the classical DLVO theory. The results elucidates the underlying physics of filtration.

Isolation of microorganisms using sub-micrometer constrictions.

We present an automated method for isolating pure bacterial cultures from samples containing multiple species that exploits the cell's own physiology to perform the separation. Cells compete to reach a chamber containing nutrients via a constriction whose cross-sectional area only permits a single cell to enter, thereby blocking the opening and preventing other cells from entering. The winning cell divides across the constriction and its progeny populate the chamber. The devices are passive and require no user interaction to perform their function. Device fabrication begins with the creation of a master mold that contains the desired constriction and chamber features. Replica molding is used to create patterned polymer chips from the master, which are bonded to glass microscope cover slips to create the constrictions. We tested constriction geometries ranging from 500 nanometers to 5 micrometers in width, 600 to 950 nanometers in height, and 10 to 40 micrometers in length. The devices were used to successfully isolate a pure Pseudomonas aeruginosa culture from a mixture that also contained Escherichia coli. We demonstrated that individual strains of the same species can be separated out from mixtures using red and green fluorescently-labeled E. coli. We also used the devices to isolate individual environmental species. Roseobacter sp. was separated from another marine species, Psychroserpens sp.

Using surface plasmon resonance imaging to study bacterial biofilms.

This paper describes the use of Surface Plasmon Resonance imaging (SPRi) as an emerging technique to study bacterial physiology in real-time without labels. The overwhelming majority of bacteria on earth exist in large multicellular communities known as biofilms. Biofilms are especially problematic because they facilitate the survival of pathogens, leading to chronic and recurring infections as well as costly industrial complications. Monitoring biofilm accumulation and removal is therefore critical in these and other applications. SPRi uniquely provides label-free, high-resolution images of biomass coverage on large channel surfaces up to 1 cm(2) in real time, which allow quantitative assessment of biofilm dynamics. The rapid imaging capabilities of this technique are particularly relevant for multicellular bacterial studies, as these cells can swim several body lengths per second and divide multiple times per hour. We present here the first application of SPRi to image Escherichia coli and Pseudomonas aeruginosa cells moving, attaching, and forming biofilms across a large surface. This is also the first time that biofilm removal has been visualized with SPRi, which has important implications for monitoring the biofouling and regeneration of fluidic systems. Initial images of the removal process show that the biofilm releases from the surface as a wave along the direction of the fluid flow.