Digital Imaging as a Detection Method for a Fluorescent Protease Assay in 96-Well and Miniaturized Assay Plate Formats
| Autor: | Matthew A. Sills, Peter Fuerst, Laura Abriola, Robert Schweitzer, Michael H. Chin |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
0301 basic medicine
business.industry Computer science Digital imaging Nanotechnology 01 natural sciences Biochemistry Fluorescence Signal 0104 chemical sciences Analytical Chemistry Cost savings 010404 medicinal & biomolecular chemistry 03 medical and health sciences 030104 developmental biology Miniaturization Molecular Medicine business Throughput (business) Computer hardware Plate reader Biotechnology |
| Zdroj: | Journal of biomolecular screening. 4(3) |
| ISSN: | 1552-454X |
| Popis: | The demand to increase throughput in HTS programs, without a concomitant addition to costs, has grown significantly during the past few years. One approach to handle this demand is assay miniaturization, which can provide greater throughput, as well as significant cost savings through reduced reagent costs. Currently, one of the major challenges facing assay miniaturization is the ability to detect the assay signal accurately and rapidly in miniaturized formats. Digital imaging is a detection method that can measure fluorescent or luminescent signals in these miniaturized formats. In this study, an imaging system capable of detecting the signal from a fluorescent protease assay in multiple plate formats was used to evaluate this detection method in an HTS environment. A direct comparison was made between the results obtained from the imaging system and a fluorescent plate reader by screening 8,800 compounds in a 96-well plate format. The imaging system generated similar changes in relative signal for each well in the screen, identified the same active compounds, and yielded similar IC(50) values as compared to the plate reader. When a standard protease inhibitor was evaluated in 96-, 384-, 864-, and 1536-well plates using imaging detection, similar IC(50) values were obtained. Furthermore, similar dose-response curves were generated for the compound in 96- and 384-well assay plates read in a plate reader. These results provide support for digital imaging as an accurate and rapid detection method for high-density microtiter plates. |
| Databáze: | OpenAIRE |
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