Development of fluorescence-based high-throughput screening assays: choice of appropriate instrumentation

David J. Burns; Elisabeth Alder; Yi-Hong Fan; Evelyn McKeegan; Usha Warrior; Bruce Beutel

doi:10.1117/12.307328

10 April 1998 Development of fluorescence-based high-throughput screening assays: choice of appropriate instrumentation

David J. Burns, Elisabeth Alder, Yi-Hong Fan, Evelyn McKeegan, Usha Warrior, Bruce Beutel

Author Affiliations +

Proceedings Volume 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery; (1998) https://doi.org/10.1117/12.307328
Event: BiOS '98 International Biomedical Optics Symposium, 1998, San Jose, CA, United States

Abstract

Fluorescence-based assays have become increasingly popular in high throughput screening for a variety of reasons (e.g. sensitivity). However, new screening technologies are pushing the limits of conventional fluorescence plate readers. For example, instruments that have optical sensitivities beyond most of the commercially available plate readers are required to reproducibly measure the fluorescence generated by the green fluorescent protein (GFP)--a novel reporter gene. Also, miniaturization of screening formats (with densities higher than the conventional 96-well plate) requires high resolution instrumentation to measure fluorescence. Several assays based on optical fluorescence measurements have been developed and screened in our Biological Screening group. These assays include various fluorescence-based protease assays (standard end-point and kinetic modes) and a functional cell-based screen using the green fluorescent protein as a reporter gene. The choice of instrumentation was the critical factor in the performance and success of each of these arrays. Data will be presented for the cell- based reporter assay including the type of instrumentation (fluorescence plate readers; fluorescence imaging systems) used for detection of GFP fluorescence.

Citation Download Citation

David J. Burns, Elisabeth Alder, Yi-Hong Fan, Evelyn McKeegan, Usha Warrior, and Bruce Beutel "Development of fluorescence-based high-throughput screening assays: choice of appropriate instrumentation", Proc. SPIE 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery, (10 April 1998); https://doi.org/10.1117/12.307328

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