A new principle of cell sorting by using selective electroporation in a modified flow cytometer

Area of Science:

• Biotechnology
• Cell Biology
• Electrical Engineering

Background:

• Electroporation involves applying electric fields to induce temporary or permanent pores in cell membranes.
• High field strengths or long pulse durations can lead to irreversible membrane damage and cell death.
• Traditional cell sorting methods can be complex and time-consuming.

Purpose of the Study:

• To develop a modified flow cytometer for electroporating individual cells selected by optical analysis.
• To utilize selective cell killing via electroporation as a method for cell sorting.
• To assess the efficiency and purity of cell sorting using this technique.

Main Methods:

• A modified flow cytometer was developed, integrating optical analysis with electroporation capabilities.
• Hydrodynamically focused cells were optically analyzed, and unwanted cells were subjected to electric pulses (10 µs, 2.0-3.2 x 10^6 V/m) in a Coulter orifice.
• Selective killing of K562 cells and lymphocytes was performed, followed by separation of viable cells.

Main Results:

• The system demonstrated the feasibility of sorting by selective killing using electric fields.
• Cell sorting rates of 1,000 cells/s were achieved.
• A high purity of the sorted fraction, reaching 99.9%, was obtained.

Conclusions:

• This modified flow cytometry system offers a simple yet effective method for cell sorting through selective electroporation and killing.
• The technique allows for high-throughput and high-purity isolation of desired cells.
• This approach has significant potential for various cell separation applications.