title: Real-time cytotoxicity assays.
abstract: Validation of new therapeutic targets calls for the advance in innovative assays that probe both spatial and temporal relationships in signaling networks. Cell death assays have already found a widespread use in pharmacological profiling of anticancer drugs. Such assays are, however, predominantly restricted to end point DEAD/LIVE parameter that provides only a snapshot of inherently stochastic process such as tumor cell death. Development of new methods that can offer kinetic real-time analysis would be highly advantageous for the pharmacological screening and predictive toxicology. In the present work we outline innovative protocols for the real-time analysis of tumor cell death, based on propidium iodide (PI) and SYTOX Green probes. These can be readily adapted to both flow cytometry and time-lapse fluorescence imaging. Considering vast time savings and kinetic data acquisition such assays have the potential to be applied in a number of areas including accelerated anticancer drug discovery and high-throughput screening routines.
authors.affiliation: The BioMEMS Research Group, Department of Chemistry, University of Auckland, Auckland, New Zealand. d.wlodkowic@elec.gla.ac.uk
authors.forename: Donald
authors.initials: D
authors.lastname: Wlodkowic
authors.forename: Shannon
authors.initials: S
authors.lastname: Faley
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Darzynkiewicz
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Cooper
mesh_terms: D000818:Animals; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor / Q000295:instrumentation / Q000379:methods*; D005434:Flow Cytometry; D006801:Humans; D057054:Molecular Imaging; D013997:Time Factors