Next Article in Journal
Homeostasis as the Mechanism of Evolution
Previous Article in Journal
Multi-Facial, Non-Peptidic α-Helix Mimetics
Article Menu

Export Article

Open AccessArticle
Biology 2015, 4(3), 556-572; doi:10.3390/biology4030556

The Impact of Photobleaching on Microarray Analysis

Institute of Technical Chemistry, Leibniz University Hanover, Callinstr. 5, 30167 Hanover, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Chris O’Callaghan
Received: 29 June 2015 / Revised: 20 August 2015 / Accepted: 8 September 2015 / Published: 11 September 2015
View Full-Text   |   Download PDF [1017 KB, uploaded 11 September 2015]   |  

Abstract

DNA-Microarrays have become a potent technology for high-throughput analysis of genetic regulation. However, the wide dynamic range of signal intensities of fluorophore-based microarrays exceeds the dynamic range of a single array scan by far, thus limiting the key benefit of microarray technology: parallelization. The implementation of multi-scan techniques represents a promising approach to overcome these limitations. These techniques are, in turn, limited by the fluorophores’ susceptibility to photobleaching when exposed to the scanner’s laser light. In this paper the photobleaching characteristics of cyanine-3 and cyanine-5 as part of solid state DNA microarrays are studied. The effects of initial fluorophore intensity as well as laser scanner dependent variables such as the photomultiplier tube’s voltage on bleaching and imaging are investigated. The resulting data is used to develop a model capable of simulating the expected degree of signal intensity reduction caused by photobleaching for each fluorophore individually, allowing for the removal of photobleaching-induced, systematic bias in multi-scan procedures. Single-scan applications also benefit as they rely on pre-scans to determine the optimal scanner settings. These findings constitute a step towards standardization of microarray experiments and analysis and may help to increase the lab-to-lab comparability of microarray experiment results. View Full-Text
Keywords: microarray; DNA; photobleaching; fluorophore; cyanine dye; bioinformatics; bioanalytics microarray; DNA; photobleaching; fluorophore; cyanine dye; bioinformatics; bioanalytics
Figures

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Supplementary material

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

von der Haar, M.; Preuß, J.-A.; von der Haar, K.; Lindner, P.; Scheper, T.; Stahl, F. The Impact of Photobleaching on Microarray Analysis. Biology 2015, 4, 556-572.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Biology EISSN 2079-7737 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top