An integrated plastic microfluidic device was designed and fabricated for bacterial detection and identification. The device, made from poly(cyclic olefin) with integrated graphite ink electrodes and photopatterned gel domains, accomplishes DNA amplification, microfluidic valving, sample injection, on-column labeling, and separation. Polymerase chain reaction (PCR) is conducted in a channel reactor containing a volume as small as 29 nL; thermal cycling utilizes screen-printed graphite ink resistors. In situ gel polymerization was employed to form local microfluidic valves that minimize convective flow of the PCR mixture into other regions. After PCR, amplicons (products) are electrokinetically injected through the gel valve, followed by on-chip electrophoretic separation. An intercalating dye is admixed to label the amplicons; they are detected using laser-induced fluorescence. Two model bacteria, Escherichia coli O157 and Salmonella typhimurium, were chosen to demonstrate bacterial detection and identification based on amplification of several of their unique DNA sequences. The limit of detection is about six copies of target DNA.
A novel technique for accurately obtaining nitrogen adsorption isotherms on thin porous films has been developed. These isotherms are useful for characterizing the surface area and pore size distribution of porous samples. The sensitivity to adsorbed nitrogen is increased by several orders of magnitude over conventional techniques by forming the test film on the substrate of a surface acoustic wave (SAW) device. This device functions as a microbalance able to detect less than 100 pg/cm2 of film. Surface areas and pore size distributions calculated from adsorption isotherms obtained with this technique on silicate sol-gel films are compared to those for bulk samples prepared from similar sol-gel solutions.
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