Lab-on-a-chip device for microfluidic trapping and TIRF imaging of single cells

Area of Science:

• Biophysics
• Microfluidics
• Cell Biology

Background:

• Total internal reflection fluorescence (TIRF) microscopy offers insights into cell membrane composition.
• Current TIRF methods often require chemical fixation, impacting cell viability and promoting clustering.
• There is a need for non-invasive methods to study live cell membranes using TIRF.

Purpose of the Study:

• To develop a microfluidic device for trapping single yeast cells for TIRF microscopy.
• To enable high-resolution imaging of cell membranes in viable cells.
• To optimize cell trapping for efficient and stable TIRF measurements.

Main Methods:

• Fabrication of microfluidic cell traps using two-photon polymerization.
• Design of 3D structures to encapsulate single yeast cells while maintaining flow exposure.
• Optimization of trap configuration, cell concentration, and injection methods for trapping efficiency.

Main Results:

• Achieved high trapping efficiencies for single yeast cells with minimal residual movement.
• Identified optimal trap configurations for robust cell immobilization.
• Demonstrated the adaptability of the trap design for use with soft hydrogel materials.

Conclusions:

• The developed microfluidic device facilitates non-invasive TIRF microscopy of live yeast cell membranes.
• This approach overcomes limitations of chemical fixation, preserving cell viability.
• The technology holds potential for broader applications in single-cell studies and membrane research.