Related Experiment Videos
Novel surface and multicolor charge coupled device-based fluorescent imaging system for DNA microarrays
D Che1, Y Bao, U R Müller
1Vysis, Incorporated, 3100 Woodcreek Drive, Downers Grove, IL 60559, USA. dche@illumina.com
Journal of Biomedical Optics
|December 1, 2001
Summary
Researchers developed a new DNA microarray system using a chromium-coated glass support and a charge-coupled device (CCD) imaging system. This innovative approach enhances fluorescent imaging sensitivity and reduces background noise for DNA analysis.
Area of Science:
- Biotechnology
- Materials Science
- Analytical Chemistry
Background:
- DNA microarrays are crucial for genetic analysis.
- Autofluorescence and nonspecific binding can limit microarray sensitivity.
- Existing methods often require DNA modification for attachment.
Purpose of the Study:
- To develop a novel support, attachment chemistry, and imaging system for DNA microarrays.
- To improve sensitivity and reduce background noise in DNA microarray assays.
- To enable efficient DNA fragment attachment without chemical modifications.
Main Methods:
- Utilized soda lime glass coated with chromium as a novel support material.
- Developed a concomitant attachment chemistry for DNA fragments (>300 nucleotides).
- Employed a charge-coupled device (CCD)-based imaging system with a xenon arc lamp.
Main Results:
- Chromium coating eliminated autofluorescence and reduced nonspecific binding.
- Achieved DNA attachment without chemical modification of DNA or surface.
- Demonstrated a detection limit of <5 x 10(7) fluorophors/cm(2) with 10 s integration.
- Observed a four-order-of-magnitude dose response in a model system.
Conclusions:
- The novel chromium-coated support and CCD imaging system significantly enhance DNA microarray performance.
- This system offers high sensitivity, low background, and efficient DNA attachment.
- The developed technology is suitable for genomic comparative genomic hybridization and other DNA analysis applications.