Whole-Cell Electrical Activity Under Direct Mechanical Stimulus by AFM Cantilever Using Planar Patch Clamp Chip...

Area of Science:

• Biophysics
• Nanotechnology
• Cell Biology

Background:

• Patch clamp technique is crucial for ion channel research.
• Existing planar patch clamp chips have limitations in multi-technique integration and fabrication cost.
• Need for improved planar patch clamp technology for advanced cellular studies.

Purpose of the Study:

• To design, fabricate, and characterize an improved silicon-based planar patch clamp chip.
• To enable long-term whole-cell voltage clamp recordings.
• To integrate planar patch clamp with microfluidics and atomic force microscopy for multi-parametric analysis.

Main Methods:

• Fabrication of silicon-based planar patch clamp chips using MEMS batch processes.
• Formation of giga-ohm seals with cell membranes.
• Whole-cell voltage clamp recordings on CHO and PC12 cells.
• Integration with a custom microfluidics chamber for perfusion.
• Combined planar patch clamp and AFM for mechanosensitive current recording in HaCaT cells.

Main Results:

• Achieved giga-ohm seals with success rates comparable to existing techniques.
• Enabled extended whole-cell voltage clamp recordings on various cell types.
• Demonstrated successful perfusion of extra- and intra-cellular buffers.
• Successfully recorded mechanosensitive currents by directly stimulating cells with AFM.
• Revealed spatial distribution and temporal dynamics of mechanosensitive ion channels.

Conclusions:

• The developed silicon-based planar patch clamp chip offers a cost-effective and versatile platform for ion channel research.
• Integration with microfluidics and AFM enables multi-parametric, high-throughput studies.
• The technology holds significant potential for advancing our understanding of ion channel proteins and cellular functions.