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A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence

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,...

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Detalles Bibliográficos
Autores principales: Asanov, Alexander, Zepeda, Angélica, Vaca, Luis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3304140/
https://www.ncbi.nlm.nih.gov/pubmed/22438738
http://dx.doi.org/10.3390/s120201800
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author Asanov, Alexander
Zepeda, Angélica
Vaca, Luis
author_facet Asanov, Alexander
Zepeda, Angélica
Vaca, Luis
author_sort Asanov, Alexander
collection PubMed
description 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.
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spelling pubmed-33041402012-03-21 A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence Asanov, Alexander Zepeda, Angélica Vaca, Luis Sensors (Basel) Article 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. Molecular Diversity Preservation International (MDPI) 2012-02-09 /pmc/articles/PMC3304140/ /pubmed/22438738 http://dx.doi.org/10.3390/s120201800 Text en © 2012 by the authors; licensee MDPI, Basel, Switzerland This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Asanov, Alexander
Zepeda, Angélica
Vaca, Luis
A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
title A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
title_full A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
title_fullStr A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
title_full_unstemmed A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
title_short A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
title_sort platform for combined dna and protein microarrays based on total internal reflection fluorescence
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3304140/
https://www.ncbi.nlm.nih.gov/pubmed/22438738
http://dx.doi.org/10.3390/s120201800
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