Digital Detection of Single Virus Particles by Multi-Spot,Label-Free Imaging Biosensor on Anti-Reflective Glass

Rapid detection of whole virus particles in biological or environmental samples represents an unmet need for the containment of infectious diseases. Here, an optical device enabling the enumeration of single virion particles binding on antibody or aptamers immobilized on a surface with anti-reflective coating is described. In this regime, nanoparticles adhering to the sensor surface provide localized contributions to the reflected field that become detectable because of their mixing with the interfering waves in the reflection direction. Thus, these settings are exploited to realize a scan-free, label-free, micro-array-type digital assay on a disposable cartridge, in which the virion counting takes place in wide field-of-view imaging. With this approach we could quantify, by enumeration, different variants of SARS-CoV-2 virions interacting with antibodies and aptamers immobilized on different spots. For all tested variants, the aptamers showed larger affinity but lower specificity relative to the antibodies. It is found that the combination of different probes on the same surface enables increasing specificity of detection and dynamic range.     

Silane-Mediated Expansion of Domains in Si-Doped κ-Ga2O3 Epitaxy and its Impact on the In-Plane Electronic Conduction

Unintentionally doped (001)-oriented orthorhombic κ-Ga₂O₃ epitaxial films on c-plane sapphire substrates are characterized by the presence of ≈ 10 nm wide columnar rotational domains that can severely inhibit in-plane electronic conduction. Comparing the in- and out-of-plane resistance on well-defined sample geometries, it is experimentally proved that the in-plane resistivity is at least ten times higher than the out-of-plane one. The introduction of silane during metal-organic vapor phase epitaxial growth not only allows for n-type Si extrinsic doping, but also results in the increase of more than one order of magnitude in the domain size (up to ≈ 300 nm) and mobility (highest µ ≈ 10 cm²V⁻¹s⁻¹, with corresponding lowest ρ ≈ 0.2 Ωcm). To qualitatively compare the mean domain dimension in κ-Ga₂O₃ epitaxial films, non-destructive experimental procedures are provided based on X-ray diffraction and Raman spectroscopy. The results of this study pave the way to significantly improved in-plane conduction in κ-Ga₂O₃ and its possible breakthrough in new generation electronics. The set of cross-linked experimental techniques and corresponding interpretation here proposed can apply to a wide range of material systems that suffer/benefit from domain-related functional properties.     

Assembling three one-camera images for three-camera intersection classification

Determining whether an autonomous self-driving agent is in the middle of an intersection can be extremely difficult when relying on visual input taken from a single camera. In such a problem setting, a wider range of views is essential, which drives us to use three cameras positioned in the front, left, and right of an agent for better intersection recognition. However, collecting adequate training data with three cameras poses several practical difficulties; hence, we propose using data collected from one camera to train a three-camera model, which would enable us to more easily compile a variety of training data to endow our model with improved generalizability. In this work, we provide three separate fusion methods (feature, early, and late) of combining the information from three cameras. Extensive pedestrian-view intersection classification experiments show that our feature fusion model provides an area under the curve and F1-score of 82.00 and 46.48, respectively, which considerably outperforms contemporary three- and one-camera models.     

A Fluorescence-Based Assay for N5-Carboxyaminoimidazole Ribonucleotide Mutase

The enzyme N⁵-carboxylaminoinidazole ribonucleotide (N⁵-CAIR) mutase is found in microbial de novo purine biosynthesis but is absent in humans making it an attractive antimicrobial target. N⁵-CAIR mutase catalyzes the synthesis of carboxyaminoimidazole ribonucleotide (CAIR) from N⁵-CAIR which is itself prepared from aminoimidazole ribonucleotide (AIR) by the enzyme N⁵-CAIR synthetase. During our research on identifying inhibitors of N⁵-CAIR mutase, we developed an innovative, fluorescence-based assay to measure the activity of this enzyme. This assay relies upon our recent serendipitous observation that AIR reversibly reacts with the compound isatin. Reaction of a fluorescently-tagged isatin with AIR resulted in a large increase in fluorescence intensity allowing a measurement of the concentration of AIR in solution. From this observation, we developed a reproducible, non-continuous assay that can replicate the known kinetic parameters of the enzyme and can readily detect a recognized inhibitor of the enzyme. This assay should find utility in screening for inhibitors targeting N⁵-CAIR mutase.