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Hydrogel-based protein and oligonucleotide microchips on metal-coated surfaces: Enhancement of fluorescence and optimization of immunoassay

Paper ID Volume ID Publish Year Pages File Format Full-Text
24449 43515 2009 9 PDF Available
Title
Hydrogel-based protein and oligonucleotide microchips on metal-coated surfaces: Enhancement of fluorescence and optimization of immunoassay
Abstract

Manufacturing of hydrogel-based microchips on metal-coated substrates significantly enhances fluorescent signals upon binding of labeled target molecules. This observation holds true for both oligonucleotide and protein microchips. When Cy5 is used as fluorophore, this enhancement is 8–10-fold in hemispherical gel elements and 4–5-fold in flattened gel pads, as compared with similar microchips manufactured on uncoated glass slides. The effect also depends on the hydrophobicity of metal-coated substrate and on the presence of a layer of liquid over the gel pads. The extent of enhancement is insensitive to the nature of formed complexes and immobilized probes and remains linear within a wide range of fluorescence intensities. Manufacturing of gel-based protein microarrays on metal-coated substrates improves their sensitivity using the same incubation time for immunoassay. Sandwich immunoassay using these microchips allows shortening the incubation time without loss of sensitivity. Unlike microchips with probes immobilized directly on a surface, for which the plasmon mechanism is considered responsible for metal-enhanced fluorescence, the enhancement effect observed using hydrogel-based microchips on metal-coated substrates might be explained within the framework of geometric optics.

Keywords
Hydrogel-based protein and oligonucleotide microarrays; Metal-coated and glass substrates; Enhancement of fluorescence signal; Comparison of surface and gel-based microarrays; Quantitative immunoassay
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Hydrogel-based protein and oligonucleotide microchips on metal-coated surfaces: Enhancement of fluorescence and optimization of immunoassay
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 144, Issue 2, 26 October 2009, Pages 151–159
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
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