Daniel R. Coughlin

Surround optical fiber immunoassay (SOFIA): an ultra-sensitive assay for prion protein detection.

We describe the development of a new technology (SOFIA) and demonstrate its utility by establishing a sensitive and specific assay for PrP(Sc). SOFIA is a surround optical fiber immunoassay which is comprised of a set of specific monoclonal antibodies and comprehensive capture of high energy fluorescence emission. In its current format, this system is capable of detecting less than 10 attogram (ag) of hamster, sheep and deer recombinant PrP. Approximately 10 ag of PrP(Sc) from 263 K-infected hamster brains can be detected with similar lower limits of PrP(Sc) detection from the brains of scrapie-infected sheep and deer infected with chronic wasting disease. These detection limits allow protease treated and untreated material to be diluted beyond the point where PrP(C), non-specific proteins or other extraneous material may interfere with PrP(Sc) signal detection and/or specificity. This not only eliminates the issue of specificity of PrP(Sc) detection but also increases sensitivity since the possibility of partial PrP(Sc) proteolysis is no longer a concern. SOFIA will likely lead to early antemortem detection of transmissible encephalopathies and is also amenable for use with additional target amplification protocols. SOFIA represents a sensitive means for detecting specific proteins involved in disease pathogenesis and/or diagnosis that extends beyond the scope of the transmissible spongiform encephalopathies.

Imaging the Rapid Solidification of Metallic Alloys in the TEM

The macroscopic properties of a metal solidified from a liquid melt are strongly dependent on the final microstructure, which in turn is the result of the solidification conditions. With the growing popularity of laser-based additive manufacturing (AM), there is an increasing need to understand the microstructures that result from rapid solidification processes. Rapidly solidified alloy microstructures are typically far from equilibrium and therefore traditional thermodynamic approaches used to predict structure and composition (i.e., phase diagrams) must be extended to describe these deviations from equilibrium and ensuing metastable states. This work highlights progress toward corroborating predictive (phase-field) modeling capabilities [1] with in situ experimental observations [2] in order to better understand the non-equilibrium structures produced during rapid solidification following laser melting.