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A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
TIRF Technologies, 951 Aviation Parkway, Suite 700, Morrisville, NC 27560, USA
Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad Universitaria, DF 04510, México
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad Universitaria, DF 04510, México
* Authors to whom correspondence should be addressed.
Received: 9 December 2011; in revised form: 13 January 2012 / Accepted: 2 February 2012 / Published: 9 February 2012
Abstract: We have developed a novel microarray technology based on total internal reflection fluorescence (TIRF) in combination with DNA and protein bioassays immobilized at the TIRF surface. Unlike conventional microarrays that exhibit reduced signal-to-background ratio, require several stages of incubation, rinsing and stringency control, and measure only end-point results, our TIRF microarray technology provides several orders of magnitude better signal-to-background ratio, performs analysis rapidly in one step, and measures the entire course of association and dissociation kinetics between target DNA and protein molecules and the bioassays. In many practical cases detection of only DNA or protein markers alone does not provide the necessary accuracy for diagnosing a disease or detecting a pathogen. Here we describe TIRF microarrays that detect DNA and protein markers simultaneously, which reduces the probabilities of false responses. Supersensitive and multiplexed TIRF DNA and protein microarray technology may provide a platform for accurate diagnosis or enhanced research studies. Our TIRF microarray system can be mounted on upright or inverted microscopes or interfaced directly with CCD cameras equipped with a single objective, facilitating the development of portable devices. As proof-of-concept we applied TIRF microarrays for detecting molecular markers from Bacillus anthracis, the pathogen responsible for anthrax.
Keywords: TIRF; molecular beacon (MB); point-of-care diagnostics (POCD); protein microarrays; DNA microarrays
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Asanov, A.; Zepeda, A.; Vaca, L. A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence. Sensors 2012, 12, 1800-1815.
Asanov A, Zepeda A, Vaca L. A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence. Sensors. 2012; 12(2):1800-1815.
Asanov, Alexander; Zepeda, Angélica; Vaca, Luis. 2012. "A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence." Sensors 12, no. 2: 1800-1815.