A microfluidic platform for whole-membrane integrity profiling in live neuronal cells.

Area of Science:

• Cell Biology
• Neuroscience
• Biophysics

Background:

• Cellular membrane integrity is crucial for health and disease, particularly in neurodegenerative disorders like Parkinson's disease.
• Current methods for assessing membrane integrity are limited in dynamic, whole-cell, and single-cell measurements.
• There is a need for advanced techniques to monitor membrane integrity over time across the entire cell surface.

Purpose of the Study:

• To develop and apply a microfluidic platform for real-time, label-free assessment of membrane integrity.
• To investigate the effects of different aggregated forms of alpha-Synuclein (aSyn) on neuronal membranes.
• To enable dynamic, single-cell, whole-membrane analysis of membrane disruption.

Main Methods:

• A microfluidic platform integrating electrokinetic microdevices with 3D microelectrodes and imaging was developed.
• The system allows continuous analysis of live neuronal cells in flow.
• Electrorotation responses were measured to quantify changes in plasma membrane capacitance.

Main Results:

• The platform successfully assessed membrane integrity in real-time and label-free.
• Different aggregated forms of alpha-Synuclein (monomeric, oligomeric, fibrillar) exhibited distinct membrane-disruptive effects.
• The study achieved single-cell resolution and whole-membrane sensitivity in quantifying these effects.

Conclusions:

• The developed microfluidic platform offers a powerful tool for studying membrane integrity in cellular models.
• This method provides a time-resolved comparison of the membrane-disruptive potential of various alpha-Synuclein conformations.
• The findings contribute to understanding the role of alpha-Synuclein in Parkinson's disease pathogenesis at the membrane level.