Evolution of a magnetic-based biomolecular detection system.

Amongst the plethora of affinity biosensor systems based on biomolecular recognition and labeling assays, magnetic labeling and detection has emerged as a promising approach. Magnetic labels can be detected by a wide range of non-invasive methods, are physically and chemically stable, relatively inexpensive to produce, and can be easily made biocompatible. Over a decade ago, the U. S. Naval Research Laboratory pioneered the use of giant magnetoresistive (GMR) sensors to detect biomolecules labeled with paramagnetic microbeads. Since then, our various investigations and engineering efforts have resulted in significant improvements in both the magnetoelectronic instrumentation and the assays associated with these magnetic labels. This paper and subsequent presentation provides a synopsis of the development of our technology which has evolved into a highly sensitive detection method.

Particle tracking single protein-functionalized quantum dot diffusion and binding at silica surfaces.

We evaluate commercial QD585 and QD605 streptavidin-functionalized quantum dots (QDs) for single-particle tracking microscopy at surfaces using total internal reflectance fluorescence and measure single QD diffusion and nonspecific binding at silica surfaces in static and flow conditions. The QD diffusion coefficient on smooth, near-ideal, highly hydroxylated silica surfaces is near bulk-solution diffusivity, as expected for repulsive surfaces, but many QD trajectories on rougher, less-than-ideal surfaces or regions display transient adsorptions. We attribute the binding to defect sites or adsorbates, possibly in conjunction with protein conformation changes, and estimate binding energies from the transient adsorption lifetimes. We also assess QD parameters relevant to tracking, including hydrodynamic radius, charge state, signal levels, blinking reduction with reducing solutions, and photoinduced blueing and bleaching.