Multiparameter Lab-on-a-Chip flow cytometry of the cell cycle

Area of Science:

• Biomedical Engineering
• Cell Biology
• Analytical Chemistry

Background:

• Understanding anticancer drug mechanisms requires analyzing apoptosis in relation to cell cycle position.
• Conventional flow cytometry (FCM) is a standard tool for such analyses, but advancements in microfluidic technology offer potential for miniaturization and improved efficiency.

Purpose of the Study:

• To demonstrate a novel application of microfluidic Lab-on-a-Chip flow cytometry (μFCM) for multiparameter analysis of apoptosis and DNA ploidy in human hematopoietic cancer cells.
• To validate the use of a simple 2D hydrodynamic focusing system within a microfluidic device for accurate cellular measurements.
• To present μFCM as a viable alternative to conventional FCM for studying programmed cell death.

Main Methods:

• Fabrication of disposable microfluidic cartridges using injection molding from optically transparent poly(methylmethacrylate).
• Integration of miniaturized electronic hardware with spatially separated solid-state lasers (473 nm and 640 nm) for up to six-parameter detection.
• Utilizing a simple 2D hydrodynamic focusing technique for cellular analysis within the chip-based device.

Main Results:

• The μFCM system successfully measured cellular DNA content in both fixed and living tumor cells.
• Demonstrated feasibility of multiparameter analysis of caspase activation and mitochondrial membrane potential (ΔΨ(m) loss) correlated with DNA content.
• Acquired high-quality biological data using straightforward microfluidic chip designs coupled with sophisticated electronic interfaces.

Conclusions:

• Simple microfluidic chip designs, when combined with advanced electronic interfaces, are sufficient for high-quality biological data acquisition.
• Microfluidic Lab-on-a-Chip flow cytometry (μFCM) provides a viable and potentially more accessible alternative to conventional FCM for multiparameter detection of programmed cell death.
• This μFCM approach facilitates the exploration of anticancer drug mechanisms by enabling detailed analysis of apoptosis and cell cycle interactions.