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Published on: August 1, 2017
Label-free prehybridization DNA microarray imaging using photonic crystals for quantitative spot quality analysis
S George1, I D Block, S I Jones
1Department of Bioengineering, University of Illinois at Urbana-Champaign, 1304 West Springfield Avenue, Urbana, Illinois 61801, USA.
Analytical Chemistry
|September 23, 2010
Summary
Label-free imaging using photonic crystal biosensors improves microarray quality control. This method identifies missed spots and assesses feature quality, enhancing data reliability in experiments.
Area of Science:
- Biotechnology
- Biosensing
- Microarray Technology
Background:
- Technical variability in DNA probe printing hinders microarray data quality.
- Current visualization methods often require fluorescent labels, altering standard protocols.
Purpose of the Study:
- To develop a label-free technique for assessing microarray spot quality before hybridization.
- To utilize photonic crystal biosensors for high-resolution imaging of printed microarrays.
Main Methods:
- Employed a photonic crystal biosensor and label-free imaging instrument.
- Generated prehybridization images of spotted oligonucleotide microarrays.
- Quantified spot intensity, size, saturation, and background for automated quality assessment.
Main Results:
- Achieved >95% sensitivity in identifying missed spots.
- Developed a spot quality scoring system to reflect feature integrity.
- Filtering by quality scores improved correlation of posthybridization data between replicate arrays.
Conclusions:
- Label-free imaging provides valuable prehybridization quality control for microarrays.
- This technique enhances data reproducibility by addressing printing variability.
- Applicable to various microarray types, including cDNA, oligonucleotide, and protein arrays.
