Electroporation is the application of controlled, pulsed electric fields to biological systems. If the biological system contains a lipid bilayer, such as is the case if the system is a suspension of cells or liposomes, the pulsed electric field may overcome the field potential of the lipid bilayer, resulting in a reversible breakdown of the bilayer and a resulting formation of temporal pores in the membrane. The pores formed are of the order of 40 to 120 nm. Most pores reseal within a few seconds, after allowing the transfer of materials into and out of the cells.
During a typical electroporation process, target cells and molecules are mixed together. When an electroporation pulse is delivered, the result is the formation of temporal pores. Before the pores reseal, the target molecules are observed to enter the cells. Upon resealing of the pores, the molecules become incorporated within the cell. The eventual target site depends on the application; for example, molecules can remain in the cytoplasm, interact with the membrane, and move into the nucleus.
Applications for electroporation include permeabilization of virtually all cells to a wide variety of molecules and ions. The most common applications for electroporation are the transformation or transfection of cells with DNA or RNA. Other applications for electroporation include electroactivation, electroinsertion of proteins into cell membranes and electroextraction of molecules from cells. Although electroporation has mainly been used as a research tool, recent work has demonstrated its potential for clinical applications. Some areas being explored include:
Cell or protoplast fusion may occur during the electroporation process, if the cells/protoplasts are brought into physical contact prior to the delivery of the pulsed electric field. In the electrofusion process, cells/protoplasts may be brought into contact through an alternating current (AC) alignment. The AC current causes a dielectrophoresis, which may result in the formation of pearl chains of cells/protoplasts. After delivery of the direct current (DC) pulse, pores that have been formed in close juxtaposition may reseal upon one another. If the process results in an intact hybrid, electrofusion has occurred. Commonly, a second round of AC alignment is employed following resealing, in an attempt to compress or stabilize the hybrid, and increasing the efficiency of the process.
Applications for electrofusion include: animal cloning, animal nuclear transfer, animal embryo manipulation, hybridoma formation, and transgenic plant production.
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