Chip-based dynamic real-time quantification of drug-induced cytotoxicity in human tumor cells

Wlodkowic, D., Skommer, J., McGuinness, D., Faley, S., Kolch, W., Darzynkiewicz, Z. and Cooper, J.M. (2009) Chip-based dynamic real-time quantification of drug-induced cytotoxicity in human tumor cells. Analytical Chemistry, 81(16), pp. 6952-6959. (doi: 10.1021/ac9010217)

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Publisher's URL: http://dx.doi.org/10.1021/ac9010217

Abstract

Cell cytotoxicity tests are among the most common bio-assays using flow cytometry and fluorescence imaging analysis. The permeability of plasma membranes to charged fluorescent probes serves, in these assays, as a marker distinguishing live from dead cells. Since it is generally assumed that probes, such as propidium iodide (PI) or 7-amino-actinomycin D (7-AAD), are themselves cytotoxic, they are currently generally used only as the end-point markers of assays for live versus dead cells. In the current study, we provide novel insights into potential applications of these classical plasma membrane integrity markers in the dynamic tracking of drug-induced cytotoxicity. We show that treatment of a number of different human tumor cell lines in cultures for up to 72 h with the PI, 7-AAD, SYTOX Green (SY-G), SYTOX Red (SY-R), TO-PRO, and YO-PRO had no effect on cell viability assessed by the integrity of plasma membrane, cell cycle progression, and rate of proliferation. We subsequently explore the potential of dynamic labeling with these markers in real-time analysis, by comparing results from both conventional cytometry and microfluidic chips. Considering the simplicity of the staining protocols and their low cost combined with the potential for real-time data collection, we show how that real-time fluorescent imaging and Lab-on-a-Chip platforms have the potential to be used for automated drug screening routines.

Item Type:Articles
Keywords:7-amino-actinomycin-d, apoptosis, assay, assays, cells, chip, chromatin, culture, cultures, cytometry, death, flow-cytometry, integrity, lab-on-a-chip, laser flow-cytometry, lines, low-cost, necrosis, plasma, progression, proliferation, propidium, protein, syto probes, tracking
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Cooper, Professor Jonathan and McGuinness, Dr Dagmara
Authors: Wlodkowic, D., Skommer, J., McGuinness, D., Faley, S., Kolch, W., Darzynkiewicz, Z., and Cooper, J.M.
Subjects:R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
College/School:College of Science and Engineering > School of Engineering > Biomedical Engineering
College of Science and Engineering > School of Engineering > Electronics and Nanoscale Engineering
Journal Name:Analytical Chemistry
Journal Abbr.:Anal. chem.
ISSN:0003-2700
ISSN (Online):1520-6882
Published Online:02 July 2009
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