Faculty Collaboration Database

Novel fluorescent technology platform for high throughput cytotoxicity and proliferation assays. J Biomol Screen 2000 Jun;5(3):141-52

Date

07/14/2000

Pubmed ID

10894757

DOI

10.1177/108705710000500306

Scopus ID

2-s2.0-0033945450 (requires institutional sign-in at Scopus site)   90 Citations

Abstract

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.

Author List

Wodnicka M, Guarino RD, Hemperly JJ, Timmins MR, Stitt D, Pitner JB

Author

Magdalena Chrzanowska PhD Associate Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Anti-Bacterial Agents
Biosensing Techniques
Cell Division
Cell Line
Fluorescent Dyes
Humans
Microbial Sensitivity Tests
Oxygen
Spectrometry, Fluorescence