Fluorescence readouts in HTS: no gain without pain?
Fluorescence-based detection technologies are frequently applied in biological testing, due to their unique advantages in setting up homogeneous, sensitive assays in miniaturized formats. However, the wide application of these readouts has highlighted challenges in reagent design and problems with interference from biological reagents and compounds. Here, we summarize the current application of fluorescence-based detection methodologies, focusing on the problems faced by assay developers and on solutions to reduce false positive and negative results in fluorescence-based HTS.
Gribbon
Philip
P
Lead Discovery Technologies, Pfizer Global Research and Development, Ramsgate Road, Sandwich, UK CT13 9NJ. philip_gribbon@sandwich.pfizer.com
Sewing
Andreas
A
D002148:Calmodulin-Binding Proteins / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation / Q000379:methods*; D029721:Drosophila Proteins; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005188:False Negative Reactions; D005189:False Positive Reactions; D005453:Fluorescence; D013678:Technology, Pharmaceutical / Q000295:instrumentation / Q000379:methods
D002148:Calmodulin-Binding Proteins; D029721:Drosophila Proteins; C108208:hts protein, Drosophila