Sonia E. Létant

Study of porous glass doped with quantum dots or laser dyes under alpha irradiation

We demonstrate that nanocomposite materials based on semiconductor quantum dots have potential for radiation detection via scintillation. While quantum dots and laser dyes both emit in the visible range at room temperature, the Stokes shift of the dyes is significantly larger. The scintillation output of both systems was studied under alpha irradiation and interpreted using a combination of energy loss and photon transport Monte Carlo simulation models. The comparison of the two systems, which allows the quantification of the role played by the Stokes shift in the scintillation output, opens up exciting possibilities for a new class of scintillators that would take advantage of the limitless assembly of nanocrystals in large, transparent, and sturdy matrices.

Polystyrene Particles Reveal Pore Substructure As They Translocate

In this article, we report resistive-pulse sensing experiments with cylindrical track-etched PET pores, which reveal that the diameters of these pores fluctuate along their length. The resistive pulses generated by polymer spheres passing through these pores have a repeatable pattern of large variations corresponding to these diameter changes. We show that this pattern of variations enables the unambiguous resolution of multiple particles simultaneously in the pore, that it can detect transient sticking of particles within the pore, and that it can confirm whether any individual particle completely translocates the pore. We demonstrate that nonionic surfactant has a significant impact on particle velocity, with the velocity decreasing by an order of magnitude for a similar increase in surfactant concentration. We also show that these pores can differentiate by particle size and charge, and we explore the influence of electrophoresis, electroosmosis, and pore size on particle motion. These results have practical importance for increasing the speed of resistive-pulse sensing, optimizing the detection of specific analytes, and identifying particle shapes.

Multiplexed Reverse Transcriptase PCR Assay for Identification of Viral Respiratory Pathogens at the Point of Care

ABSTRACT We have developed a nucleic acid-based assay that is rapid, sensitive, and specific and can be used for the simultaneous detection of five common human respiratory pathogens, including influenza virus A, influenza virus B, parainfluenza virus types 1 and 3, respiratory syncytial virus (RSV), and adenovirus groups B, C, and E. Typically, diagnosis on an unextracted clinical sample can be provided in less than 3 h, including sample collection, preparation, and processing, as well as data analysis. Such a multiplexed panel would enable rapid broad-spectrum pathogen testing on nasal swabs and therefore allow implementation of infection control measures and the timely administration of antiviral therapies. We present here a summary of the assay performance in terms of sensitivity and specificity. The limits of detection are provided for each targeted respiratory pathogen, and result comparisons were performed on clinical samples, our goal being to compare the sensitivity and specificity of the multiplexed assay to the combination of immunofluorescence and shell vial culture currently implemented at the University of California-Davis Medical Center hospital. Overall, the use of the multiplexed reverse transcription-PCR assay reduced the rate of false-negative results by 4% and reduced the rate of false-positive results by up to 10%. The assay correctly identified 99.3% of the clinical negatives and 97% of the adenovirus, 95% of the RSV, 92% of the influenza virus B, and 77% of the influenza virus A samples without any extraction performed on the clinical samples. The data also showed that extraction will be needed for parainfluenza virus, which was only identified correctly 24% of the time on unextracted samples.

Hydrogen Production by a Hyperthermophilic Membrane-Bound Hydrogenase in Water-Soluble Nanolipoprotein Particles

Hydrogenases constitute a promising class of enzymes for ex vivo hydrogen production. Implementation of such applications is currently hindered by oxygen sensitivity and, in the case of membrane-bound hydrogenases (MBHs), poor water solubility. Nanolipoprotein particles (NLPs) formed from apolipoproteins and phospholipids offer a novel means of incorporating MBHs into a well-defined water-soluble matrix that maintains the enzymatic activity and is amenable to incorporation into more complex architectures. We report the synthesis, hydrogen-evolving activity, and physical characterization of the first MBH-NLP assembly. This may ultimately lead to the development of biomimetic hydrogen-production devices.

New method for attachment of biomolecules to porous silicon.

Biomolecules have been attached to porous silicon by a new linking method that forms a direct Si-C bond on the surface and retains the photoluminescence of the porous silicon.

Rapid-Viability PCR Method for Detection of Live, Virulent Bacillus anthracis in Environmental Samples

ABSTRACT In the event of a biothreat agent release, hundreds of samples would need to be rapidly processed to characterize the extent of contamination and determine the efficacy of remediation activities. Current biological agent identification and viability determination methods are both labor- and time-intensive such that turnaround time for confirmed results is typically several days. In order to alleviate this issue, automated, high-throughput sample processing methods were developed in which real-time PCR analysis is conducted on samples before and after incubation. The method, referred to as rapid-viability (RV)-PCR, uses the change in cycle threshold after incubation to detect the presence of live organisms. In this article, we report a novel RV-PCR method for detection of live, virulent Bacillus anthracis, in which the incubation time was reduced from 14 h to 9 h, bringing the total turnaround time for results below 15 h. The method incorporates a magnetic bead-based DNA extraction and purification step prior to PCR analysis, as well as specific real-time PCR assays for the B. anthracis chromosome and pXO1 and pXO2 plasmids. A single laboratory verification of the optimized method applied to the detection of virulent B. anthracis in environmental samples was conducted and showed a detection level of 10 to 99 CFU/sample with both manual and automated RV-PCR methods in the presence of various challenges. Experiments exploring the relationship between the incubation time and the limit of detection suggest that the method could be further shortened by an additional 2 to 3 h for relatively clean samples.

Hydrolysis of acetylcholinesterase inhibitors--organophosphorus acid anhydrolase enzyme immobilization on photoluminescent porous silicon platforms.

We report on the immobilization of an OPAA enzyme on luminescent porous silicon devices, and on the utilization of this new platform to hydrolyze p-nitrophenyl-soman.

Most-Probable-Number Rapid Viability PCR method to detect viable spores of Bacillus anthracis in swab samples

A comparison of Most-Probable-Number Rapid Viability (MPN RV) PCR and traditional culture methods for the quantification of Bacillus anthracis Sterne spores in macrofoam swabs from a multi-center validation study was performed. The purpose of the study was to compare environmental swab processing methods for recovery, detection, and quantification of viable B. anthracis spores from surfaces. Results show that spore numbers provided by the MPN RV-PCR method were typically within 1-log of the values from a plate count method for all three levels of spores tested (3.1x10(4), 400, and 40 spores sampled from surfaces with swabs) even in the presence of debris. The MPN method tended to overestimate the expected result, especially at lower spore levels. Blind negative samples were correctly identified using both methods showing a lack of cross contamination. In addition to detecting low levels of spores in environmental conditions, the MPN RV-PCR method is specific, and compatible with automated high-throughput sample processing and analysis protocols, enhancing its utility for characterization and clearance following a biothreat agent release.

Line Edge Detection and Characterization in SEM Images Using Wavelets

Edge characterization has become increasingly important in nanotechnology due to the growing demand for precise nanoscale structure fabrication and assembly. Edge detection and assembly. Edge detection is often performed by thresholding the spatial information of a top-down image obtained by scanning electron microscopy or other surface characterization techniques. Results are highly dependent on an arbitrary threshold value, which makes it difficult to reveal the nature of the real surface and to compare results among images. In this paper, we present an alternative edge boundary detection technique based on the wavelet framework. Our results indicate that the method facilitates nanoscale edge detection and characterization by providing a systematic threshold determination step.

Detection of bio-organism simulants using random binding on a defect-free photonic crystal

The defect-free photonic crystal (PC) slab geometry was explored for size-selective detection of bio-organism simulants. Through feedback between finite-difference time-domain simulations and experiments, we generated a conservative limit of detection estimate for randomized pore filling of a two-dimensional PC slab, and predict that random binding affords the label-free PC-based optical detection of low numbers (of the order of 10) of biological particles.