Velocity measurement of particulate flow in microfluidic channels using single point confocal fluorescence detection.

This article presents a non-invasive, optical technique for measuring particulate flow within microfluidic channels. Confocal fluorescence detection is used to probe single fluorescently labeled microspheres (0.93 microm diameter) passing through a focused laser beam at a variety of flow rates (50 nL min(-1)-8 microL min(-1)). Simple statistical methods are subsequently used to investigate the resulting fluorescence bursts and generate velocity data for the flowing particles. Fluid manipulation is achieved by hydrodynamically pumping fluid through microchannels (150 microm wide and 50 microm deep) structured in a polydimethylsiloxane (PDMS) substrate. The mean fluorescence burst frequency is shown to be directly proportional to flow speed. Furthermore, the Poisson recurrence time and width of recovered autocorrelation curves is demonstrated to be inversely proportional to flow speed. The component-based confocal fluorescence detection system is simple and can be applied to a diversity of planar chip systems. In addition, velocity measurement only involves interrogation of the fluidic system at a single point along the flow stream, as opposed to more normal multiple-point measurements.

Production of monoclonal antibodies to viruses in the potyvirus group: use in radioimmunoassay.

Hybridomas were developed that secreted antibodies to soybean mosaic (SMV), lettuce mosaic (LMV) viruses, and to maize dwarf mosaic (MDMV) virus, strains Ap and B. All hybridomas produced antibodies specific to the homologous virus except for one to LMV, which produced antibodies that reacted at a low level with SMV and MDMV. Monoclonal antibodies against SMV were used in a double-antibody sandwich radioimmunoassay. The assay using only one type of monoclonal antibody lacked sensitivity because, presumably, limited epitopes were available. Results obtained using two monoclonal antibodies that bound to different epitopes were comparable to those using polyclonal antibodies. A competitive radioimmunoassay, using a single monoclonal antibody, was developed to detect successfully SMV, LMV and MDMV.