Dr. Jay A. West Profile

Dr. Jay A. West

Senior Member Technical Staff at Sandia National Labs California

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Proceedings Article | 8 December 2004 Paper

Microfluidic gene arrays for rapid genomic profiling

Jay West, Kyle Hukari, Gary Hux, Timothy Shepodd

Proceedings Volume 5591, (2004) https://doi.org/10.1117/12.578619

KEYWORDS: Microfluidics, Polymers, Molecules, Profiling, Charge-coupled devices, Target detection, Signal detection, Polymer thin films, Robotics, Chemical elements

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Genomic analysis tools have recently become an indispensable tool for the evaluation of gene expression in a variety of experiment protocols. Two of the main drawbacks to this technology are the labor and time intensive process for sample preparation and the relatively long times required for target/probe hybridization. In order to overcome these two technological barriers we have developed a microfluidic chip to perform on chip sample purification and labeling, integrated with a high density genearray. Sample purification was performed using a porous polymer monolithic material functionalized with an oligo dT nucleotide sequence for the isolation of high purity mRNA. These purified mRNA’s can then rapidly labeled using a covalent fluorescent molecule which forms a selective covalent bond at the N7 position of guanine residues. These labeled mRNA’s can then released from the polymer monolith to allow for direct hybridization with oligonucletide probes deposited in microfluidic channel. To allow for rapid target/probe hybridization high density microarray were printed in microchannels. The channels can accommodate array densities as high as 4000 probes. When oligonucleotide deposition is complete, these channels are sealed using a polymer film which forms a pressure tight seal to allow sample reagent flow to the arrayed probes. This process will allow for real time target to probe hybridization monitoring using a top mounted CCD fiber bundle combination. Using this process we have been able to perform a multi-step sample preparation to labeled target/probe hybridization in less than 30 minutes. These results demonstrate the capability to perform rapid genomic screening on a high density microfluidic microarray of oligonucleotides.

Proceedings Article | 8 December 2004 Paper

Rapid universal solublization and analysis of bacterial spores using a simple flow-through ultrahigh-temperature capillary device

Jay West, Kyle Hukari, Ronald Renzi, Kamlesh Patel

Proceedings Volume 5591, (2004) https://doi.org/10.1117/12.578623

KEYWORDS: Capillaries, Proteins, Statistical analysis, Chemical analysis, Microfluidics, Biological research, Viruses, Bacteria, Luminescence, Temperature metrology

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Rapid identification of viral and bacterial species is dependent of the ability to manipulate biological agents into a form where they are directly analyzed. Many of these species of interest, such as bacterial spores, are inherently hearty and very difficult to lyse or solubilize. Standard protocols for spore inactivation include, chemical treatment, sonication, pressure and thermal lysis. While these protocols are effective for the inactivation of these agents they are less well suited for sample preparation for analysis using capillary electrophoresis techniques. In order to overcome this difficulty we fabricated a simple capillary device to perform thermal lysis of vegatative bacterial cells and bacterial spores. Using an ethylene glycol buffer to super heat bacterial spores we were able to perform rapid flow through lysis and solubilzation of these agents. This device was then coupled to a sample preparation station for on-line fluorescamine dye lableling and buffer exchange for direct analysis using a miniaturized capillary electrophoresis instrument. Using this integrated device were we enabled to perform sample lysis, labeling and protein fingerprint analysis of vegatative bacterial cells, bacterial spores and viruses in less than 10 minutes. The described device is simple, inexpensive and easily integratable with various microfluidic devices.

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