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Positioning of the sensor cell on the sensing area using cell trapping pattern in incubation type planar patch clamp
Zhi-Hong Wang1, Noriko Takada, Hidetaka Uno
1Nagoya University, Nagoya 464-8603, Japan.
Colloids and Surfaces. B, Biointerfaces
|April 25, 2012
Summary
A novel cell trapping pattern on sensor chips improves planar patch clamp biosensor operation. This pattern facilitates cell monolayer formation over micropores, enabling accurate channel current measurements.
Area of Science:
- Biosensor technology
- Cell biology
- Electrophysiology
Background:
- Incubation-based planar patch clamp biosensors face challenges in precise sensor cell positioning and stable incubation.
- Achieving a confluent cell monolayer over the sensor's micropore is crucial for reliable measurements but difficult to obtain.
Purpose of the Study:
- To develop and evaluate a novel cell trapping pattern for enhanced planar patch clamp biosensor performance.
- To improve sensor cell adhesion, positioning, and monolayer formation for accurate electrophysiological recordings.
Main Methods:
- Fabrication of a silicon sensor chip with a micropore-centered, 5 μm deep lattice cell trapping pattern.
- Coating the chip surface with extracellular matrix collagen IV and seeding HEK293 cells expressing channel-rhodopsin-wide-receiver (ChR-WR).
- Utilizing the nystatin perforation technique for whole-cell arrangement and laser stimulation (473 nm) for channel current induction.
Main Results:
- The cell trapping pattern successfully confined cells within the designated area, promoting rapid monolayer formation over the micropore.
- In contrast, flat sensor chips without the pattern showed delayed cell coverage and formation of multi-cell layers, hindering measurements.
- Laser-induced channel currents were successfully measured from the cell monolayer, with characteristics consistent with traditional pipette patch clamp methods.
Conclusions:
- The developed cell trapping pattern significantly enhances the operational efficiency and reliability of incubation-type planar patch clamp biosensors.
- This approach overcomes key limitations in cell positioning and monolayer formation, paving the way for more robust biosensing applications.
- The results demonstrate the pattern's effectiveness in facilitating accurate electrophysiological measurements using whole-cell patch clamp configurations.
