title: A modular, fully integrated ultra-high-throughput screening system based on confocal fluorescence analysis techniques.
abstract: The rapid increase in size of compound libraries, as well as new targets emerging from the Human Genome Project, require progress in ultra-high-throughput screening (uHTS) systems. In a joint effort with scientists and engineers from the biotech and the pharmaceutical industry, a modular, fully integrated system for miniaturized uHTS was developed. The goal was to achieve high data quality in small assay volumes (1-4 microL) combined with reliable and unattended operation. Two new confocal fluorescence readers have been designed. One of the instruments is a 4-channel confocal fluorescence reader, measuring with 4 objectives in parallel. The fluorescence readout is based on single-molecule detection methods, allowing high sensitivity at low tracer concentrations and delivering an information-rich output. The other instrument is a confocal fluorescence imaging reader, where the images are analyzed in terms of generic patterns and quantified in units of intensity per pixel. Both readers are spanning the application range from assays with isolated targets in homogenous solution or membrane vesicle-based assays (4-channel reader) to cell-based assays (imaging reader). Results from a comprehensive test on these assay types demonstrate the high quality and robustness of this screening system.
authors.affiliation: Evotec OAI/Evotec Technologies, Hamburg, Germany. stefan.jaeger@evotecoai.com
authors.forename: Stefan
authors.initials: S
authors.lastname: Jäger
authors.forename: Norbert
authors.initials: N
authors.lastname: Garbow
authors.forename: Achim
authors.initials: A
authors.lastname: Kirsch
authors.forename: Hartwig
authors.initials: H
authors.lastname: Preckel
authors.forename: Frank U
authors.initials: FU
authors.lastname: Gandenberger
authors.forename: Kurt
authors.initials: K
authors.lastname: Herrenknecht
authors.forename: Martin
authors.initials: M
authors.lastname: Rüdiger
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Hutchinson
authors.forename: Ryan P
authors.initials: RP
authors.lastname: Bingham
authors.forename: Fernando
authors.initials: F
authors.lastname: Ramon
authors.forename: Ana
authors.initials: A
authors.lastname: Bardera
authors.forename: Julio
authors.initials: J
authors.lastname: Martin
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D016923:Cell Death; D002460:Cell Line; D002470:Cell Survival; D003201:Computers; D004340:Drug Contamination; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation* / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D020128:Inhibitory Concentration 50; D008024:Ligands; D018613:Microscopy, Confocal; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis; D012270:Ribosomes / Q000187:drug effects; D012680:Sensitivity and Specificity; D020298:U937 Cells
chemical_list: D000900:Anti-Bacterial Agents; D008024:Ligands; D010455:Peptides; D011506:Proteins