SUMMARY
The discussion centers on the discrepancy in fluorescence readings between a cuvette and a 96 well plate in a fluorometric assay. The user reports successful fluorescence detection using a cuvette but not with a black, flat-bottomed, opaque 96 well plate from Greiner. Key factors affecting the results include the excitation and emission wavelengths (Exc. 468 nm, Em. 572 nm), the materials of the cuvette (fused silica, borosilicate glass, or quartz), and the sensitivity of the fluorometer used. It is concluded that plate readers are generally less sensitive than traditional cuvette fluorometers, which may contribute to the observed differences in fluorescence.
PREREQUISITES
- Understanding of fluorometry and fluorescence principles
- Familiarity with 96 well plate specifications and materials
- Knowledge of excitation and emission wavelengths
- Experience with different types of fluorometers and their configurations
NEXT STEPS
- Investigate the differences in sensitivity between cuvette fluorometers and 96 well plate readers
- Research the impact of well plate materials on fluorescence readings
- Learn about the optimal positioning of detectors in fluorometric assays
- Explore the specifications and performance of various fluorometers, particularly those designed for 96 well plates
USEFUL FOR
Researchers and laboratory technicians conducting fluorescence assays, particularly those using fluorometers and 96 well plates for bio-assays. This discussion is beneficial for anyone troubleshooting fluorescence detection issues in their experiments.