Paper-fluidic devices have become an emerging trend for micro total analysis systems (microTAS) in the bioengineering field due to their ability to maintain the rapid, sensitive and specific attributes of microfluidic devices. Subsequently, paper-fluidic devices are also more portable, have a lower production cost and are easier to use. However, one of the obstacles in developing paper fluidic devices is the limited ability to control the rate of fluid flow during an assay. In our project, we use electrowetting on dielectrics where a dielectric, which is normally hydrophobic, is polarized and becomes hydrophilic. We have fabricated paper-fluidic devices by inkjet printing and spraying conductive hydrophobic electrodes/valves in conjunction with conductive hydrophilic electrodes which are able to stop the fluid front of phosphate buffered saline (PBS). The hydrophobic valves were then actuated by an applied potential which altered the fluorinated monolayer on the electrode. As the applied potential between the electrodes was increased, the amount of time for the fluid front to pass the valve decreased because the monolayer was altered faster. However, we did not observe significant differences in time as we increased the distance between the electrodes. The valves were also incorporated in a lateral flow assay where the device was used to detect Saccharomyces cerevisiae rRNA sequences. With the ability to control the fluid flow in a paper-fluidic device, more complex and intricate assays can be developed, which not only can be applied in the biomedical, food and environmental fields, but also can be used in low resource settings for the detection of diseases.
Bacteriophages are viruses capable of infecting and lysing target bacterial cells; as such they have potential applications in agriculture for decontamination of foods, food contact surfaces and food rinse water. Although bacteriophages can retain infectivity long-term using lyophilized storage, the process of freeze-drying can be time consuming and expensive. In this study, electrospinning was used for dehydrating bacteriophages in polyvinylpyrrolidone polymer solutions with addition of excipients (sodium chloride, magnesium sulfate, Tris-HCl, sucrose) in deionized water. The high voltage dehydration reduced the infectivity of bacteriophages following electrospinning, with the damaging effect abated with addition of storage media (SM) buffer and sucrose. SM buffer and sucrose also provided the most protection over extended storage (8 weeks; 20 °C; 1% relative humidity) by mitigating environmental effects on the dried bacteriophages. Magnesium sulfate however provided the least protection due to coagulation effects of the ion, which can disrupt the native conformation of the bacteriophage protein coat. Storage temperatures (20 °C, 4 °C and -20 °C; 1% relative humidity) had a minimal effect while relative humidity had substantial effect on the infectivity of bacteriophages. Nanofibers stored in higher relative humidity (33% and 75%) underwent considerable damage due to extensive water absorption and disruption of the fibers. Overall, following storage of nanofiber mats for eight weeks at ambient temperatures, high infective phage concentrations (10-10 PFU ml) were retained. Therefore, this study provided valuable insights on preservation and dehydration of bacteriophages by electrospinning in comparison to freeze drying and liquid storage, and the influence of excipients on the viability of bacteriophages.
Antisolvent precipitation is commonly used to fabricate protein nanoparticles using a simple batch method that involves injecting a protein-solvent mixture into an antisolvent. In this study, the potential of producing core-shell protein nanoparticles by antisolvent precipitation using a continuous dual-channel microfluidization method was investigated. The solvent phase (zein in ethanol) and antisolvent phase (casein in water) were made to impinge on each other at high velocity, which generates intense shear, turbulent, and cavitation forces that ensure thorough mixing and breakup of the phases. Relatively small core-shell protein nanoparticles (d<125nm) could be produced using this method when the conditions were optimized. The mean particle diameter decreased with increasing antisolvent-to-solvent ratio, increasing homogenization pressure, increasing ethanol content in the solvent phase, and decreasing zein content in the solvent phase. Depending on the processing conditions employed, zein particles in the range of about 120nm to over 1000nm could be produced. The operating conditions were further optimized to increase the final zein concentration and decrease the organic solvent content while still obtaining small particles. The surface potential of the core-shell protein nanoparticles went from positive at low pH to negative at high pH, with a point of zero charge around pH5. Electron microscopy indicated that the protein particles formed had a roughly spherical shape. The results suggest that the dual-channel microfluidizer could be used to continuously form protein nanoparticles by antisolvent precipitation. Nevertheless, when the microfluidization method was compared with the simple batch method the size of the particles produced under similar conditions were fairly similar.
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