Although the large-scale analysis of the human genome has provided a wealth of information for the genetic analysis of cancer and other diseases, most of these advances are unavailable in the clinic due to their expense and complexity. The development of miniaturized devices capable of automated real time analysis of genetic profiles is likely to enable routine genetic analysis of diseases such as cancer, whether for diagnosis or for monitoring treatment throughout the course of the disease. Microfluidic chips allow detection of mutations and abnormal gene expression patterns. Here, we describe the application of microfluidic chips for the molecular monitoring of gene expression profiles associated with human cancer. On-chip RT-PCR products are detectable after as few as 15 cycles of PCR, and from individual cells. On-chip detection is as sensitive as or exceeds the sensitivity obtained using conventional technologies.
"However, without on-chip valves it was difficult to control the flow of the PCR mixture and integrate the PCR module with additional on-chip analysis systems. Our work with non-flowing PCR within a well or internal reservoir (Backhouse et al., 2003; Pilarski et al., 2004) suffered low yields because of the tendency of the PCR mix to dry out or move under the influence of thermophoretic forces and/or the pressure caused by the release of dissolved gasses. "
[Show abstract][Hide abstract] ABSTRACT: On-chip genetic analysis systems are beginning to provide a viable alternative to conventional gene profiling and amplification devices, through minimal reagent use, high detection resolution, and the potential for high-throughput parallel testing of the genetic material, even from single cells. Despite the advantages, there are many difficulties inherent in creating an integrated microfluidic diagnostic platform. One major challenge is the accurate control and manipulation of fluid, and particularly the immobilization of reaction mixtures during heating phases of polymerase chain reactions (PCR). In this paper we present a pumping and valving system based on the use of three servomotor-controlled valve fingers that actuate microchannels within a poly-dimethylsiloxane (PDMS) fluidic chip. We characterize the valving ability of the system in terms of fluid loss and show the successful fluid retention of the system over 35-cycle PCR runs at temperatures of up to approximately 96 degrees C. In addition, we demonstrate the system's ability to perform PCR by successfully amplifying a sample of beta2 microglobulin transcript obtained from the peripheral blood of a patient with multiple myeloma. This work has proven to be a successful approach to multi-use valving and a viable method of alleviating the fluid control difficulties inherent in performing a PCR reaction in an on-chip environment. In addition, it opens the door for further automation and integration with other chip-based genetic analysis platforms.
"The development of microfluidic platforms for the use in lab-on-chip devices has the potential to reveal detailed health information through automated real-time analysis (Backhouse et al., 2003). Just as large computers have been decreased to the size of dime-sized chips, so too have the tools of biotechnology undergone drastic miniaturization (Staeder, 2002). "
[Show abstract][Hide abstract] ABSTRACT: We present a method to assess microchip performance for on-chip electrophoretic separations. The assessment is realized through electrophoretic manipulation of a DNA size standard using specially designed electric voltage programs. Results achieved from the assessment could be used as an indicator of microchip "aging" in terms of lowered resolutions and fluctuations in electro-osmotic flow (EOF).
MEMS, NANO and Smart Systems, 2004. ICMENS 2004. Proceedings. 2004 International Conference on; 09/2004
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