Publications

Throckmorton, Daniel J.; Helm, Jonathan I.; Warmouth, John A.; Van De Vreugde, James L.; Gilbert, Jolene R.

Abstract not provided.

Jebrail, Mais J.; Sinha, Anupama S.; Renzi, Ronald F.; Van De Vreugde, James L.; Gondhalekar, Carmen G.; Schoeniger, Joseph S.; Branda, Steven B.

Abstract not provided.

PLoS ONE

Bartsch, Michael B.; Edwards, Harrison S.; Lee, Daniel; Moseley, Caroline E.; Tew, Karen E.; Renzi, Ronald F.; Van De Vreugde, James L.; Kim, Hanyoup; Knight, Daniel L.; Sinha, Anupama S.; Branda, Steven B.; Patel, Kamlesh P.

Advances in molecular biology, microfluidics, and laboratory automation continue to expand the accessibility and applicability of these methods beyond the confines of conventional, centralized laboratory facilities and into point of use roles in clinical, military, forensic, and field-deployed applications. As a result, there is a growing need to adapt the unit operations of molecular biology (e.g., aliquoting, centrifuging, mixing, and thermal cycling) to compact, portable, low-power, and automation-ready formats. Here we present one such adaptation, the rotary zone thermal cycler (RZTC), a novel wheel-based device capable of cycling up to four different fixed-temperature blocks into contact with a stationary 4-microliter capillarybound sample to realize 1-3 second transitions with steady state heater power of less than 10 W. We demonstrate the utility of the RZTC for DNA amplification as part of a highly integrated rotary zone PCR (rzPCR) system that uses low-volume valves and syringe-based fluid handling to automate sample loading and unloading, thermal cycling, and between-run cleaning functionalities in a compact, modular form factor. In addition to characterizing the performance of the RZTC and the efficacy of different online cleaning protocols, we present preliminary results for rapid single-plex PCR, multiplex short tandem repeat (STR) amplification, and second strand cDNA synthesis.

Lab on a Chip

Jebrail, Mais J.; Renzi, Ronald F.; Sinha, Anupama S.; Van De Vreugde, James L.; Gondhalekar, Carmen; Ambriz, Cesar; Meagher, Robert M.; Branda, Steven B.

Digital microfluidics (DMF) is a powerful technique for sample preparation and analysis for a broad range of biological and chemical applications. In many cases, it is desirable to carry out DMF on an open surface, such that the matrix surrounding the droplets is ambient air. However, the utility of the air-matrix DMF format has been severely limited by problems with droplet evaporation, especially when the droplet-based biochemical reactions require high temperatures for long periods of time. We present a simple solution for managing evaporation in air-matrix DMF: just-in-time replenishment of the reaction volume using droplets of solvent. We demonstrate that this solution enables DMF-mediated execution of several different biochemical reactions (RNA fragmentation, first-strand cDNA synthesis, and PCR) over a range of temperatures (4-95°C) and incubation times (up to 1 h or more) without use of oil, humidifying chambers, or off-chip heating modules. Reaction volumes and temperatures were maintained roughly constant over the course of each experiment, such that the reaction kinetics and products generated by the air-matrix DMF device were comparable to those of conventional benchscale reactions. This simple yet effective solution for evaporation management is an important advance in developing air-matrix DMF for a wide variety of new, high-impact applications, particularly in the biomedical sciences.

Branda, Steven B.; Jebrail, Mais J.; Sinha, Anupama S.; Renzi, Ronald F.; Bartsch, Michael B.; Van De Vreugde, James L.; Gondhalekar, Carmen G.; Amriz, Cesar A.; Schoeniger, Joseph S.; Meagher, Robert M.; Patel, Kamlesh P.

Abstract not provided.

Goldsmith, John E.; Brennan, James S.; Gerling, Mark D.; Kiff, Scott D.; Mascarenhas, Nick M.; Van De Vreugde, James L.

We have developed a mobile fast neutron imaging platform to enhance the capabilities of emergency responders in the localization and characterization of special nuclear material. This mobile imager of neutrons for emergency responders (MINER) is based on the Neutron Scatter Camera, a large segmented imaging system that was optimized for large-area search applications. Due to the reduced size and power requirements of a man-portable system, MINER has been engineered to fit a much smaller form factor, and to be operated from either a battery or AC power. We chose a design that enabled omnidirectional (4π) imaging, with only a ~twofold decrease in sensitivity compared to the much larger neutron scatter cameras. The system was designed to optimize its performance for neutron imaging and spectroscopy, but it does also function as a Compton camera for gamma imaging. This document outlines the project activities, broadly characterized as system development, laboratory measurements, and deployments, and presents sample results in these areas. Additional information can be found in the documents that reside in WebPMIS.

PLoS ONE

Kim, Hanyoup; Jebrail, Mais J.; Sinha, Anupama S.; Bent, Zachary B.; Solberg, Owen D.; Williams, Kelly P.; Langevin, Stanley A.; Renzi, Ronald F.; Van De Vreugde, James L.; Meagher, Robert M.; Schoeniger, Joseph S.; Lane, Todd L.; Branda, Steven B.; Bartsch, Michael B.; Patel, Kamlesh D.

Next-generation sequencing (NGS) is emerging as a powerful tool for elucidating genetic information for a wide range of applications. Unfortunately, the surging popularity of NGS has not yet been accompanied by an improvement in automated techniques for preparing formatted sequencing libraries. To address this challenge, we have developed a prototype microfluidic system for preparing sequencer-ready DNA libraries for analysis by Illumina sequencing. Our system combines droplet-based digital microfluidic (DMF) sample handling with peripheral modules to create a fully-integrated, sample-in library-out platform. In this report, we use our automated system to prepare NGS libraries from samples of human and bacterial genomic DNA. E. coli libraries prepared on-device from 5 ng of total DNA yielded excellent sequence coverage over the entire bacterial genome, with >99% alignment to the reference genome, even genome coverage, and good quality scores. Furthermore, we produced a de novo assembly on a previously unsequenced multi-drug resistant Klebsiella pneumoniae strain BAA-2146 (KpnNDM). The new method described here is fast, robust, scalable, and automated. Our device for library preparation will assist in the integration of NGS technology into a wide variety of laboratories, including small research laboratories and clinical laboratories. © 2013 Kim et al.

Brennan, James S.; Marleau, Peter M.; Mengesha, Wondwosen M.; Mrowka, Stanley M.; Le Galloudec, Nathalie J.; Throckmorton, Daniel J.; Van De Vreugde, James L.; Tewell, Craig R.

Abstract not provided.

Branda, Steven B.; Jebrail, Mais J.; Van De Vreugde, James L.; Langevin, Stanley A.; Bent, Zachary B.; Curtis, Deanna J.; Lane, Pamela L.; Carson, Bryan C.; La Bauve, Elisa L.; Patel, Kamlesh P.; Ricken, James B.; Schoeniger, Joseph S.; Solberg, Owen D.; Williams, Kelly P.; Misra, Milind; Powell, Amy J.; Pattengale, Nicholas D.; May, Elebeoba E.; Lane, Todd L.; Lindner, Duane L.; Young, Malin M.; VanderNoot, Victoria A.; Thaitrong, Numrin T.; Bartsch, Michael B.; Renzi, Ronald F.; Tran-Gyamfi, Mary B.; Meagher, Robert M.

Abstract not provided.

Branda, Steven B.; Jebrail, Mais J.; Van De Vreugde, James L.; Langevin, Stanley A.; Bent, Zachary B.; Curtis, Deanna J.; Lane, Pamela L.; Carson, Bryan C.; La Bauve, Elisa L.; Patel, Kamlesh P.; Ricken, James B.; Schoeniger, Joseph S.; Solberg, Owen D.; Williams, Kelly P.; Misra, Milind; Powell, Amy J.; Pattengale, Nicholas D.; May, Elebeoba E.; Lane, Todd L.; Lindner, Duane L.; Young, Malin M.; VanderNoot, Victoria A.; Thaitrong, Numrin T.; Bartsch, Michael B.; Renzi, Ronald F.; Tran-Gyamfi, Mary B.; Meagher, Robert M.

Abstract not provided.

Branda, Steven B.; Jebrail, Mais J.; Van De Vreugde, James L.; Langevin, Stanley A.; Bent, Zachary B.; Curtis, Deanna J.; Lane, Pamela L.; Carson, Bryan C.; La Bauve, Elisa L.; Patel, Kamlesh P.; Ricken, James B.; Schoeniger, Joseph S.; Solberg, Owen D.; Williams, Kelly P.; Misra, Milind; Powell, Amy J.; Pattengale, Nicholas D.; May, Elebeoba E.; Lane, Todd L.; Lindner, Duane L.; Young, Malin M.; VanderNoot, Victoria A.; Thaitrong, Numrin T.; Bartsch, Michael B.; Renzi, Ronald F.; Tran-Gyamfi, Mary B.; Meagher, Robert M.

Abstract not provided.

Kim, Hanyoup; Renzi, Ronald F.; Patel, Kamlesh P.; Van De Vreugde, James L.; Thaitrong, Numrin T.; Bartsch, Michael B.

Abstract not provided.

Journal of Laboratory Automation

Kim, Hanyoup; Bartsch, Michael B.; Renzi, Ronald F.; He, Jim; Van De Vreugde, James L.; Claudnic, Mark R.; Patel, Kamlesh D.

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.

Kim, Hanyoup; Renzi, Ronald F.; Thaitrong, Numrin T.; Van De Vreugde, James L.; Bartsch, Michael B.

Abstract not provided.

Kiff, Scott D.; Wessendorf, Kurt O.; Brennan, James S.; Detry, Richard J.; Gerling, Mark D.; Kershaw, Christopher P.; Renzi, Ronald F.; Savignon, Daniel J.; Steele, John T.; Van De Vreugde, James L.

Abstract not provided.

Ferko, Scott M.; Patel, Kamlesh P.; Van De Vreugde, James L.; Haroldsen, Brent L.

Abstract not provided.

Davalos, Rafael V.; Krafcik, Karen L.; Salmi, Allen J.; Van De Vreugde, James L.; Mosier, Bruce P.; Morales, Alfredo M.

Microsystems pose unparalleled opportunity in the realm of real-time sample analysis for multiple applications, including Homeland Security monitoring devices, environmental monitoring, and biomedical diagnostics. The need for a universal means of processing, separating, and delivering a sample within these devices is a critical need if these systems are to receive widespread implementation in the industry and government sectors. Efficient particle separation and enrichment techniques are critical for a range of analytical functions including pathogen detection, sample preparation, high-throughput particle sorting, and biomedical diagnostics. Previously, using insulator-based dielectrophoresis (iDEP) in microfluidic glass devices, we demonstrated simultaneous particle separation and concentration. As an alternative to glass, we evaluate the performance of similar iDEP structures produced in polymer-based microdevices and their enhancement through dynamic surface coatings. There are numerous processing and operational advantages that motivate our transition to polymers such as the availability of numerous innate chemical compositions for tailoring performance, mechanical robustness, economy of scale, and ease of thermoforming and mass manufacturing. The polymer chips we have evaluated are fabricated through an injection molding process of the commercially available cyclic olefin copolymer Zeonor{reg_sign}. We demonstrate that the polymer devices achieve the same performance metrics as glass devices. Additionally, we show that the nonionic block copolymer surfactant Pluronic F127 has a strong interaction with the cyclic olefin copolymer at very low concentrations, positively impacting performance by decreasing the magnitude of the applied electric field necessary to achieve particle trapping. The presence of these dynamic surface coatings, therefore, lowers the power required to operate such devices and minimizes Joule heating. The results of this study demonstrate that polymeric microfluidic devices with surfactant coatings for insulator-based dielectrophoresis provide an affordable engineering strategy for selective particle enrichment and sorting.