Electroporation is a physical transfection method that uses an electrical pulse to create temporary pores in cell membranes through which substances like nucleic acids can pass into cells. It is a highly efficient strategy for the introduction of foreign nucleic acids into many cell types, including bacteria and mammalian cells.
How Electroporation Works
Electroporation is based on a simple process. Host cells and selected molecules are suspended in a conductive solution, and an electrical circuit is closed around the mixture. An electrical pulse at an optimized voltage and only lasting a few microseconds to a millisecond is discharged through the cell suspension. This disturbs the phospholipid bilayer of the membrane and results in the formation of temporary pores. The electric potential across the cell membrane simultaneously rises to allow charged molecules like DNA to be driven across the membrane through the pores in a manner similar to electrophoresis (Shigekawa and Dower, 1988).
The main advantage of electroporation is its applicability for transient and stable transfection of all cell types. Furthermore, because electroporation is easy and rapid, it is able to transfect a large number of cells in a short time once optimum electroporation conditions are determined. The major drawback of electroporation is substantial cell death caused by high voltage pulses and only partially successful membrane repair, requiring the use of greater quantities of cells compared to chemical transfection methods. While more modern instrumentation, such as the Neon® Transfection System offered by Life Technologies™, overcome high cell mortality by distributing the electrical pulse equally among the cells and maintaining a stable pH throughout the electroporation chamber, optimization of pulse and field strength parameters is still required to balance the electroporation efficiency and cell viability.
Prepare cells by suspending in electroporation buffer.
Apply electrical pulse to cells in the presence of specilized buffer and nucleic acids.
Electrical pulse creates a potential difference across the cell membrane and induces temporary pores in the membrane for nucleic acid entry.
Return cells to growth conditions and allow them to recover.
Assay cells for gene expression or silencing.
Neon® Transfection System for Electroporation
|The Neon® Transfection System is a second-generation transfection system that uses an electronic pipette tip as an electroporation chamber. The design and performance of the Neon® electronic pipette transfection chamber results in increased cell viability and transfection efficiency compared to traditional cuvette-based electroporation systems.|
The Neon® Transfection System has helped many researchers improve their transfection experiments. The system was designed for electroporation of mammalian cells, but some customers have found it to be successful for other cell types such as insect cell cultures and parasites.
The advantages of this method are:
- Versatility: Electroporation with the Neon® system is effective with mammalian cell types
- Efficiency: A high percentage of cells are transfected without jeopardizing viability
- Flexibility: A range of cell numbers can be used: from 104 to 106 cells
- Simplicity: Just one kit for all cell types
- Novel: The Neon® system employs a unique electroporation chamber, as easy to use as a pipette and with a circular shape that provides advantages over cuvette-based systems.
The design of the electroporation chamber distributes the current equally among the cells and maintains a stable pH throughout the chamber; these key benefits increase cell viability dramatically.
- Learn more about The Neon® Transfection System