Apoptotic cell death dynamics of HL60 cells studied using a microfluidic cell trap device

Area of Science:

• Biomedical Engineering
• Cell Biology
• Biophysics

Background:

• Apoptosis, or programmed cell death, is crucial for development and disease.
• Current methods for apoptosis analysis can be complex and lack real-time, single-cell resolution.
• Developing advanced tools is essential for understanding cell death pathways and for drug discovery.

Purpose of the Study:

• To design and fabricate a novel microfluidic cell trap device for analyzing apoptosis.
• To enable real-time monitoring of the apoptotic process at the single-cell level.
• To evaluate the device's efficacy in analyzing both chemically and electrically induced apoptosis.

Main Methods:

• Fabrication of a microfluidic silicon-glass chip for cell immobilization.
• Induction of apoptosis using chemical agents (TNF-alpha, CHX) or electric fields.
• Utilizing fluorescent dyes (FLICA, PI) for discriminating between viable, apoptotic, and necrotic cells.
• Real-time monitoring of cellular changes during apoptosis.

Main Results:

• The microfluidic chip successfully immobilized cells and allowed real-time monitoring of chemically induced apoptosis in HL60 cells.
• The device enabled tracking of the onset and progression of the cell death cascade.
• Distinguishing between apoptotic and necrotic stages was challenging for electric field-mediated cell death.

Conclusions:

• The developed microfluidic apoptosis chip is a promising tool for analyzing programmed cell death dynamics.
• The device facilitates real-time, single-cell analysis of apoptosis, particularly for chemically induced pathways.
• This work represents a significant step towards integrated microfluidic platforms for high-throughput drug screening.