CONFERENCE PROCEEDINGS
Advanced Search >
Home > Proceedings > Volume 8615 > Article
9 March 2013 Multivariate analysis of apoptotic markers versus cell cycle phase in living human cancer cells by microfluidic cytometry
Jin Akagi, Joanna Skommer, Anna Matuszek, Kazuo Takeda, Yuu Fujimura, Khashayar Khoshmanesh, Kourosh Kalantar-Zadeh, Arnan Mitchell, Rachel Errington, Paul J. Smith, Zbigniew Darzynkiewicz, Donald Wlodkowic
Author Affiliations +
Proceedings Volume 8615, Microfluidics, BioMEMS, and Medical Microsystems XI; 86150W (2013) https://doi.org/10.1117/12.2001474
Event: SPIE MOEMS-MEMS, 2013, San Francisco, California, United States
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.
© (2013) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation Download Citation
Jin Akagi, Jin Akagi, Joanna Skommer, Joanna Skommer, Anna Matuszek, Anna Matuszek, Kazuo Takeda, Kazuo Takeda, Yuu Fujimura, Yuu Fujimura, Khashayar Khoshmanesh, Khashayar Khoshmanesh, Kourosh Kalantar-Zadeh, Kourosh Kalantar-Zadeh, Arnan Mitchell, Arnan Mitchell, Rachel Errington, Rachel Errington, Paul J. Smith, Paul J. Smith, Zbigniew Darzynkiewicz, Zbigniew Darzynkiewicz, Donald Wlodkowic, Donald Wlodkowic, "Multivariate analysis of apoptotic markers versus cell cycle phase in living human cancer cells by microfluidic cytometry", Proc. SPIE 8615, Microfluidics, BioMEMS, and Medical Microsystems XI, 86150W (9 March 2013); doi: 10.1117/12.2001474; https://doi.org/10.1117/12.2001474
ACCESS THE FULL ARTICLE
PERSONAL SIGN IN
Full access may be available with your subscription
 
Forgot your username?
Forgot your password?
No SPIE account? Create an account
or Institutional Sign In via Shibboleth
PURCHASE THIS CONTENT
SUBSCRIBE TO DIGITAL LIBRARY
50 downloads per 1-year subscription
Members: $195
Non-members: $335 ADD TO CART
25 downloads per 1 - year subscription
Members: $145
Non-members: $250 ADD TO CART
PURCHASE SINGLE ARTICLE
Includes PDF, HTML & Video, when available
Members: $14.40
Non-members: $18.00 ADD TO CART
PROCEEDINGS
 8 PAGES
DOWNLOAD PDF + SAVE TO MY LIBRARY
SHARE
GET CITATION
< Previous Article
|
Next Article >
KEYWORDS
RELATED CONTENT
Subscribe to Digital Library
Receive Erratum Email Alert
Back to Top