Three-dimensional (3D) hydrodynamic focusing for continuous sampling and analysis of adherent cells

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Cell Biology

Background:

• Adherent cells pose challenges in microfluidic analysis due to surface attachment.
• Efficient single-cell sampling and lysis are crucial for accurate intracellular content detection.

Purpose of the Study:

• To develop a microfluidic device that prevents cell adhesion and enables rapid, high-throughput single-cell analysis.
• To integrate hydrodynamic focusing, cell lysis, and capillary electrophoresis for intracellular content measurement.

Main Methods:

• A cross microfluidic chip with three sheath-flow channels was designed to create a 'soft fluid wall', preventing cell adhesion.
• Three-dimensional hydrodynamic focusing guided single cells for continuous sampling and lysis.
• Capillary electrophoresis (CE) with sodium dodecyl sulfate (SDS) in sheath-flow solution enabled rapid cell lysis and separation.

Main Results:

• The device successfully prevented adherent cell attachment to the channel surface.
• Single-cell introduction rate reached up to 151 cells/min.
• The system achieved an average analysis throughput of 16-18 cells/min for reduced glutathione (GSH) and reactive oxygen species (ROS) in HepG2 cells.

Conclusions:

• The developed microfluidic system offers an effective solution for analyzing intracellular content in adherent cells.
• The integrated approach ensures high-throughput, single-cell analysis with minimal cell loss and rapid lysis.
• This technology has potential applications in various fields of cell biology and diagnostics.