Publications

Abstract not provided.

Abstract not provided.

Abstract not provided.

While DNA sequencing technology is advancing at an unprecedented rate, sample preparation technology still relies primarily on manual bench-top processes, which often can be slow, labor-intensive, inefficient, or inconsistent. To address these disadvantages, we developed an integrated microfluidic platform for automated preparation of DNA libraries for next generation sequencing. This sample-to-answer system has great potential for rapid characterization of novel and emerging pathogens from clinical samples.

Next-generation sequencing (NGS) technology is a promising tool for identifying and characterizing unknown pathogens, but its usefulness in time-critical biodefense and public health applications is currently limited by the lack of fast, efficient, and reliable automated DNA sample preparation methods. To address this limitation, we are developing a digital microfluidic (DMF) platform to function as a fluid distribution hub, enabling the integration of multiple subsystem modules into an automated NGS library sample preparation system. A novel capillary interface enables highly repeatable transfer of liquid between the DMF device and the external fluidic modules, allowing both continuous-flow and droplet-based sample manipulations to be performed in one integrated system. Here, we highlight the utility of the DMF hub platform and capillary interface for automating two key operations in the NGS sample preparation workflow. Using an in-line contactless conductivity detector in conjunction with the capillary interface, we demonstrate closed-loop automated fraction collection of target analytes from a continuous-flow sample stream into droplets on the DMF device. Buffer exchange and sample cleanup, the most repeated steps in NGS library preparation, are also demonstrated on the DMF platform using a magnetic bead assay and achieving an average DNA recovery efficiency of 80% ± 4.8% © 2011 Society for Laboratory Automation and Screening.

Abstract not provided.

Abstract not provided.

Abstract not provided.

Abstract not provided.

Abstract not provided.

Abstract not provided.

We have designed, fabricated, and characterized a digital microfluidic (DMF) platform to function as a central hub for interfacing multiple lab-on-a-chip sample processing modules towards automating the preparation of clinically-derived DNA samples for ultrahigh throughput sequencing (UHTS). The platform enables plug-and-play installation of a two-plate DMF device with consistent spacing, offers flexible connectivity for transferring samples between modules, and uses an intuitive programmable interface to control droplet/electrode actuations. Additionally, the hub platform uses transparent indium-tin oxide (ITO) electrodes to allow complete top and bottom optical access to the droplets on the DMF array, providing additional flexibility for various detection schemes.

Abstract not provided.

Abstract not provided.

In a multiyear research agreement with Tenix Investments Pty. Ltd., Sandia has been developing field deployable technologies for detection of biotoxins in water supply systems. The unattended water sensor or UWS employs microfluidic chip based gel electrophoresis for monitoring biological analytes in a small integrated sensor platform. This instrument collects, prepares, and analyzes water samples in an automated manner. Sample analysis is done using the {mu}ChemLab{trademark} analysis module. This report uses analysis results of two datasets collected using the UWS to estimate performance of the device. The first dataset is made up of samples containing ricin at varying concentrations and is used for assessing instrument response and detection probability. The second dataset is comprised of analyses of water samples collected at a water utility which are used to assess the false positive probability. The analyses of the two sets are used to estimate the Receiver Operating Characteristic or ROC curves for the device at one set of operational and detection algorithm parameters. For these parameters and based on a statistical estimate, the ricin probability of detection is about 0.9 at a concentration of 5 nM for a false positive probability of 1 x 10{sup -6}.

Abstract not provided.

Rapid identification of aerosolized biological agents following an alarm by particle triggering systems is needed to enable response actions that save lives and protect assets. Rapid identifiers must achieve species level specificity, as this is required to distinguish disease-causing organisms (e.g., Bacillus anthracis) from benign neighbors (e.g., Bacillus subtilis). We have developed a rapid (1-5 minute), novel identification methodology that sorts intact organisms from each other and particulates using capillary electrophoresis (CE), and detects using near-infrared (NIR) absorbance and scattering. We have successfully demonstrated CE resolution of Bacillus spores and vegetative bacteria at the species level. To achieve sufficient sensitivity for detection needs ({approx}10{sup 4} cfu/mL for bacteria), we have developed fiber-coupled cavity-enhanced absorbance techniques. Using this method, we have demonstrated {approx}two orders of magnitude greater sensitivity than published results for absorbing dyes, and single particle (spore) detection through primarily scattering effects. Results of the integrated CE-NIR system for spore detection are presented.

We present a rapid method for the identification of viruses using microfluidic chip gel electrophoresis (CGE) of high-copy number proteins to generate unique protein profiles. Viral proteins are solubilized by heating at 95°C in borate buffer containing detergent (5 min), then labeled with fluorescamine dye (10 s), and analyzed using the μChemLab CGE system (5 min). Analyses of closely related T2 and T4 bacteriophage demonstrate sufficient assay sensitivity and peak resolution to distinguish the two phage. CGE analyses of four additional viruses - MS2 bacteriophage, Epstein - Barr, respiratory syncytial, and vaccinia viruses - demonstrate reproducible and visually distinct protein profiles. To evaluate the suitability of the method for unique identification of viruses, we employed a Bayesian classification approach. Using a subset of 126 replicate electropherograms of the six viruses and phage for training purposes, successful classification with non-training data was 66/69 or 95% with no false positives. The classification method is based on a single attribute (elution time), although other attributes such as peak width, peak amplitude, or peak shape could be incorporated and may improve performance further. The encouraging results suggest a rapid and simple way to identify viruses without requiring specialty reagents such as PCR probes and antibodies. © 2008 American Chemical Society.

This document is a safety analysis of a novel neutron detection technology developed by Sandia National Laboratories. This technology is comprised of devices with tiny channels containing high pressure {sup 3}He. These devices are further integrated into large scale neutron sensors. Modeling and preliminary device testing indicates that the time required to detect the presence of special nuclear materials may be reduced under optimal conditions by several orders of magnitude using this approach. Also, these devices make efficient use of our {sup 3}He supply by making individual devices more efficient and/or extending the our limited {sup 3}He supply. The safety of these high pressure devices has been a primary concern. We address these safety concerns for a flat panel configuration intended for thermal neutron detection. Ballistic impact tests using 3 g projectiles were performed on devices made from FR4, Silicon, and Parmax materials. In addition to impact testing, operational limits were determined by pressurizing the devices either to failure or until they unacceptably leaked. We found that (1) sympathetic or parasitic failure does not occur in pressurized FR4 devices (2) the Si devices exhibited benign brittle failure (sympathetic failure under pressure was not tested) and (3) the Parmax devices failed unacceptably. FR4 devices were filled to pressures up to 4000 + 100 psig, and the impacts were captured using a high speed camera. The brittle Si devices shattered, but were completely contained when wrapped in thin tape, while the ductile FR4 devices deformed only. Even at 4000 psi the energy density of the compressed gas appears to be insignificant compared to the impact caused by the incoming projectile. In conclusion, the current FR4 device design pressurized up to 4000 psi does not show evidence of sympathetic failure, and these devices are intrinsically safe.

Abstract not provided.

Abstract not provided.

Abstract not provided.

An innovative helium3 high pressure gas detection system, made possible by utilizing Sandia's expertise in Micro-electrical Mechanical fluidic systems, is proposed which appears to have many beneficial performance characteristics with regards to making these neutron measurements in the high bremsstrahlung and electrical noise environments found in High Energy Density Physics experiments and especially on the very high noise environment generated on the fast pulsed power experiments performed here at Sandia. This same system may dramatically improve active WMD and contraband detection as well when employed with ultrafast (10-50 ns) pulsed neutron sources.

Abstract not provided.

Abstract not provided.