| Novel fluorescent technology platform for high throughput cytotoxicity and proliferation assays. | |
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MedLine Citation:
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PMID: 10894757 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We have developed a novel fluorescent Oxygen BioSensor technology platform adaptable to many applications in the area of drug discovery and development, particularly cell-based assays. This biosensor technology requires no additional reagents or incubations, and affords continuous real-time readout of dissolved oxygen concentrations. Since the level of oxygen dissolved in an assay's medium correlates to the number and viability of the cells in the medium, this technology is ideally suited for monitoring cell viability, proliferation, or death. The technology is particularly well suited to investigating cells' kinetic responses to proliferative or toxic stimuli, such as drugs. When incorporated into a 96- or 384-well microplate format, it is compatible with standard laboratory automation systems. Here we present data illustrating the application of the Oxygen BioSensor technology for rapid, homogeneous detection and evaluation of metabolic activity of a variety of eukaryotic and prokaryotic cells, including mammalian cells, insect cells, yeast, and bacteria. In the absence of toxic substances, we find a good correlation between cell number and signal over a wide range of cell concentrations and growth times. To evaluate the usefulness of the Oxygen BioSensor for cytotoxicity assays, we have performed a series of experiments using a range of toxic agents and cell types, including both bacteria and mammalian cell lines. In a side-by-side comparison to standard MTT assays using HL60 cells, comparable IC(50) values were found with the Oxygen BioSensor for five different toxins or drugs. This assay method does not have the need for additional reagents, handling steps, or incubation periods required by the MTT assays. |
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Authors:
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M Wodnicka; R D Guarino; J J Hemperly; M R Timmins; D Stitt; J B Pitner |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of biomolecular screening Volume: 5 ISSN: 1087-0571 ISO Abbreviation: J Biomol Screen Publication Date: 2000 Jun |
Date Detail:
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Created Date: 2000-08-09 Completed Date: 2000-08-09 Revised Date: 2011-05-23 |
Medline Journal Info:
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Nlm Unique ID: 9612112 Medline TA: J Biomol Screen Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 141-52 Citation Subset: IM |
Affiliation:
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BD Technologies, Research Triangle Park, NC 27709, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Anti-Bacterial Agents / pharmacology Biosensing Techniques* Cell Division / drug effects Cell Line Fluorescent Dyes Humans Microbial Sensitivity Tests Oxygen Spectrometry, Fluorescence / methods* |
| Chemical | |
Reg. No./Substance:
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0/Anti-Bacterial Agents; 0/Fluorescent Dyes; 7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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