Improved sensitivity of DNA microarrays using photonic crystal enhanced fluorescence

Patrick C. Mathias, Sarah I. Jones, Hsin Yu Wu, Fuchyi Yang, Nikhil Ganesh, Delkin O. Gonzalez, German Bollero, Lila O. Vodkin, Brian T. Cunningham

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46 Citations (Scopus)

Abstract

DNA microarrays are used to profile changes in gene expression between samples in a high-throughput manner, but measurements of genes with low expression levels can be problematic with standard microarray substrates. In this work, we expand the detection capabilities of a standard microarray experiment using a photonic crystal (PC) surface that enhances fluorescence observed from microarray spots. This PC is inexpensively and uniformly fabricated using a nanoreplica molding technique, with very little variation in its optical properties within- and between-devices. By using standard protocols to process glass microarray substrates in parallel with PCs, we evaluated the impact of this substrate on a one-color microarray experiment comparing gene expression in two developmental stages of Glycine max. The PCs enhanced the signal-to-noise ratio observed from microarray spots by 1 order of magnitude, significantly increasing the number of genes detected above substrate fluorescence noise. PC substrates more than double the number of genes classified as differentially expressed, detecting changes in expression even for low expression genes. This approach increases the dynamic range of a surface-bound fluorescence-based assay to reliably quantify small quantities of DNA that would be impossible with standard substrates.

Original languageEnglish
Pages (from-to)6854-6861
Number of pages8
JournalAnalytical Chemistry
Volume82
Issue number16
DOIs
Publication statusPublished - Aug 15 2010
Externally publishedYes

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ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Mathias, P. C., Jones, S. I., Wu, H. Y., Yang, F., Ganesh, N., Gonzalez, D. O., Bollero, G., Vodkin, L. O., & Cunningham, B. T. (2010). Improved sensitivity of DNA microarrays using photonic crystal enhanced fluorescence. Analytical Chemistry, 82(16), 6854-6861. https://doi.org/10.1021/ac100841d