Single-Cell Electroporation with Real-Time Impedance Assessment Using a Constriction Microchannel

Area of Science:

• Biomedical Engineering
• Cell Biology
• Microfluidics

Background:

• Electroporation is a key technique for intracellular delivery but faces limitations in precise electric field control and real-time monitoring.
• Current methods struggle with accurate electric field application to single cells and lack integrated real-time feedback systems.

Purpose of the Study:

• To develop a microfluidic system for precise, rapid single-cell electroporation.
• To integrate simultaneous impedance monitoring for real-time assessment of electroporation.
• To overcome limitations of conventional electroporation techniques for intracellular cargo delivery.

Main Methods:

• A microfluidic device with a constriction microchannel was designed to create a localized high electric field around single cells.
• Continuous flow of A549 cells through the microchannel allowed for electroporation as they passed the constriction.
• Impedance changes were monitored in real-time during cell passage and correlated with electroporation events.

Main Results:

• The microfluidic system achieved highly efficient single-cell electroporation, reaching up to 96.6%.
• An abrupt impedance drop upon cell entry into the constriction correlated with successful electroporation.
• Stabilized impedance measurements during cell transit quantified the extent of electroporation, validated by propidium iodide uptake.

Conclusions:

• The developed microfluidic system offers precise control over single-cell electroporation.
• Real-time, label-free impedance assessment provides a reliable metric for electroporation extent.
• This technology holds significant potential for advancing intracellular delivery and other biomedical applications.