Dynamic analysis of apoptosis using cyanine SYTO probes: From classical to microfluidic cytometry

Wlodkowic, D., Skommer, J., Faley, S., Darzynkiewicz, Z. and Cooper, J.M. (2009) Dynamic analysis of apoptosis using cyanine SYTO probes: From classical to microfluidic cytometry. Experimental Cell Research, 315(10), pp. 1706-1714. (doi:10.1016/j.yexcr.2009.03.006)

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Abstract

Cell death is a stochastic process, often initiated and/or executed in a multi-pathway/multi-organelle fashion. Therefore, high-throughput single-cell analysis platforms are required to Provide detailed characterization of kinetics and mechanisms of cell death in heterogeneous cell populations. However, there is still a largely unmet need for inert fluorescent probes, suitable for prolonged kinetic studies. Here, we compare the use of innovative adaptation of unsymmetrical SYTO dyes for dynamic real-time analysis of apoptosis in conventional as well as microfluidic chip-based systems. We show that cyanine SYTO probes allow non-invasive tracking of intracellular events over extended time. Easy handling and "stain-no wash" protocols open up new opportunities for high-throughput analysis and live-cell sorting. Furthermore, SYTO probes are easily adaptable for detection of cell death using automated microfluidic chip-based cytometry. Overall, the combined use of SYTO probes and state-of-the-art Lab-on-a-Chip platform emerges as a cost effective solution for automated drug screening compared to conventional Annexin V or TUNEL assays. In particular, it should allow for dynamic analysis of samples where low cell number has so far been an obstacle, e.g. primary cancer stems cells or circulating minimal residual tumors.

Item Type:Articles
Keywords:Antitumor drugs, apoptosis, assay, assays, cancer, cell, cell-death, cells, chip, cytometry, death, dyes, flow cytometry, kinetic assays, lab-on-a-chip, mechanisms, microfluidics, real-time, system, systems, SYTO, SYTO probes, time, tracking
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Faley, Dr Shannon and Wlodkowic, Dr Donald and Cooper, Professor Jonathan
Authors: Wlodkowic, D., Skommer, J., Faley, S., Darzynkiewicz, Z., and Cooper, J.M.
College/School:College of Science and Engineering > School of Engineering > Electronics and Nanoscale Engineering
College of Science and Engineering > School of Engineering > Biomedical Engineering
Journal Name:Experimental Cell Research
Journal Abbr.:Exp. cell res.
ISSN:0014-4827
ISSN (Online):1090-2422
Published Online:17 March 2009

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