title: Multiparameter Lab-on-a-Chip flow cytometry of the cell cycle.
abstract: Multiparameter analysis of apoptosis in relation to cell cycle position is helpful in exploring mechanism of action of anticancer drugs that target specific molecular cogs of the cell cycle. This work demonstrates a new rationale for using microfluidic Lab-on-a-Chip flow cytometry (μFCM) with a simple 2D hydrodynamic focusing for the multiparameter analysis of apoptosis and DNA ploidy analysis in human hematopoietic cancer cells. The microfluidic system employs disposable microfluidic cartridges fabricated using injection moulding in optically transparent poly(methylmethacrylate). The dedicated and miniaturized electronic hardware interface enables up to six parameter detections using a combination of spatially separated solid-state 473 nm (10 mW) and 640 nm (20 mW) lasers and x-y stage for rapid laser alignment adjustment. We provide evidence that the simple 2D flow focusing on a chip-based device is sufficient to measure cellular DNA content in both fixed and living tumor cells. The feasibility of using the μFCM system for multiparameter analysis of caspase activation and dissipation of mitochondrial inner membrane potential (ΔΨ(m) loss) in relation to DNA content is also demonstrated. The data shows that straightforward microfluidic chip designs are sufficient to acquire high quality biological data when combined with sophisticated electronic interfaces. They can be a viable alternative to conventional FCM for multiparameter detection of programmed cell death.
authors.affiliation: The BioMEMS Research Group, School of Chemical Sciences, University of Auckland, Auckland, New Zealand.
authors.forename: Joanna
authors.initials: J
authors.lastname: Skommer
authors.forename: Jin
authors.initials: J
authors.lastname: Akagi
authors.forename: Kazuo
authors.initials: K
authors.lastname: Takeda
authors.forename: Yuu
authors.initials: Y
authors.lastname: Fujimura
authors.forename: Khashayar
authors.initials: K
authors.lastname: Khoshmanesh
authors.forename: Donald
authors.initials: D
authors.lastname: Wlodkowic
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000302:isolation & purification*; D004247:DNA / Q000737:chemistry; D005434:Flow Cytometry / Q000379:methods*; D018922:HL-60 Cells; D006801:Humans; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics; D051336:Mitochondrial Membranes / Q000737:chemistry; D011003:Ploidies
chemical_list: D018797:Cell Cycle Proteins; D004247:DNA