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Multivariate analysis of apoptotic markers versus cell cycle phase in living human cancer cells by microfluidic cytometry

Becker, H., Akagi, J., Skommer, J., Matuszek, A., Takeda, K., Fujimura, Y., Khoshmanesh, K., Kalantar-Zadeh, K., Mitchell, A., Errington, Rachel Jane, Smith, Paul James, Darzynkiewicz, Z., Wlodkowic, D. and Gray, B. 2013. Multivariate analysis of apoptotic markers versus cell cycle phase in living human cancer cells by microfluidic cytometry. Proceedings of SPIE 8615 , 86150W. 10.1117/12.2001474

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Abstract

Measurement of apoptotic markers in tumors can be directly correlated with the cell cycle phase using flow cytometry (FCM). The conventional DNA content analysis requires cell permeabilization to stain nuclei with fluorescent probes such as propidium iodide or use of a costly UV-excitation line for Hoechst 33342 probe. The access to FCM is also still limited to centralized core facilities due to its inherent high costs and complex operation. This work describes development and proof-of-concept validation of a portable and user-friendly microfluidic flow cytometer (μFCM) that can perform multivariate real time analysis on live cells using sampling volumes as small as 10 microliters. The μFCM system employs disposable microfluidic cartridges fabricated using injection molding in poly(methylmethacrylate) transparent thermoplastic. Furthermore, the dedicated and miniaturized electronic hardware interface enables up to six parameter detection using a combination of spatially separated solid-state 473 (10 mW) and 640 nm (20 mW) lasers and x-y stage for rapid laser alignment adjustment. We provide new evidence that a simple 2D flow focusing on a chip is sufficient to measure cellular DNA content in live tumor cells using a far-red DNA probe DRAQ5. The feasibility of using the μFCM system for a dose-response profiling of investigational anti-cancer agents on human hematopoietic cancer cells is also demonstrated. The data show that μFCM can provide a viable novel alternative to conventional FCM for multiparameter detection of caspase activation and dissipation of mitochondrial inner membrane potential (ΔΨm) in relation to DNA content (cell cycle phase) in live tumor cells.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Subjects: R Medicine > R Medicine (General)
R Medicine > RC Internal medicine
R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
R Medicine > RZ Other systems of medicine
Uncontrolled Keywords: cancer; Cell cycle; Cell permeabilization; Miniaturized electronics; Multi variate analysis; Multi-parameter detection; Parameter detection; Programmed cell deaths
Publisher: Society of Photo-optical Instrumentation Engineers
ISSN: 0277-786X
Last Modified: 04 Jun 2017 08:44
URI: http://orca-mwe.cf.ac.uk/id/eprint/83930

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