Related Experiment Videos
Up-converting phosphor reporters for nucleic acid microarrays
F van De Rijke1, H Zijlmans, S Li
1Laboratory for Cytochemistry and Cytometry, Department of Molecular Cell Biology, Leiden University Medical Center; Leiden, The Netherlands.
Nature Biotechnology
|March 7, 2001
Summary
Up-converting phosphor technology (UPT) offers superior sensitivity for DNA microarray analysis compared to traditional fluorescent reporters. This advancement in luminescence enables more efficient RNA expression monitoring with less sample.
Area of Science:
- Biotechnology
- Molecular Biology
- Nanotechnology
Background:
- DNA microarrays are crucial for monitoring RNA expression levels.
- Conventional fluorescent reporters limit sensitivity and require large RNA amounts due to low hybridization efficiency.
Purpose of the Study:
- To evaluate up-converting phosphor technology (UPT) as a novel luminescent reporter for DNA microarrays.
- To compare the sensitivity and performance of UPT against cyanin 5 (Cy5) fluorescent reporters.
Main Methods:
- UPT utilizes unique phosphor particles that emit visible light upon infrared (IR) illumination (980 nm) via up-conversion.
- Performance was assessed by comparing UPT and Cy5 in a low-complexity model system and through cDNA microarray hybridizations.
Main Results:
- UPT demonstrated a two-order-of-magnitude linear relationship between luminescence and target concentration.
- UPT showed superior sensitivity compared to Cy5, even with standard microscopy equipment.
- Excellent correlation (R2 = 0.95) was observed between UPT and Cy5 for variable target concentrations.
Conclusions:
- UPT reporter technology significantly enhances sensitivity in DNA microarray applications.
- UPT holds promise for improving RNA expression monitoring and other microarray-based analyses.
- Combining UPT with IR laser scanners could revolutionize microarray applications.