Square Wave Electroporation
Square Wave Electroporation - Related Videos
07:23
Electroporation of Craniofacial Mesenchyme
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2011
Craniofacial cartilages develop in close contact with other tissues and are difficult to manipulate in live animals. We are using electroporation to deliver molecular tools during growth of the craniofacial skeleton while bypassing early embryonic effects. This approach will allow us to efficiently test candidate molecules in vivo.
09:30
Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection
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Cited by 49 •
2011
A gene transfer method into the developing mouse brain is described by using a unique surgical method and special shape of electrodes. This unique technique allows transfection of plasmid DNA temporally and spatially, which will aid many neuroscientists in studying brain development.
08:15
The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation
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Cited by 13 •
2009
This procedure describes how to establish primary hematopoietic cell cultures from murine bone marrow and is followed by transfection using the Gene Pulser MXCell electroporation system.
12:55
Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency
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Cited by 10 •
2010
This procedure shows how to use the Gene Pulser MXcell electroporation system to rapidly and easily identify the best electroporation conditions for mouse embryonic fibroblasts (MEFs) or other primary cells. Considerations for troubleshooting are also discussed in the associated video.
11:57
Electroporation of Mycobacteria
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Cited by 55 •
2008
Mycobacterial pathogenic strategies remain poorly understood. The slow growth rate of most species, the impenetrable nature of the cell-wall, and the hazards of working with pathogens make mycobacteria difficult to study and are largely responsible for our poor understanding of these organisms. In this video we will demonstrate the technique of electroporation, which involves subjecting cells to a brief high electrical impulse to allow the entry of DNA. It is the most widely used method for...
FAQs
A square pulse maintains a near-constant electric field rather than changing continuously during the applied interval. This field induces a transmembrane potential, the voltage difference across the membrane, strong enough to produce transient pores. The resulting permeability change creates a temporary opportunity for normally poorly permeable molecules to cross into the cell.
Pulse strength and duration determine how strongly and how long the membrane experiences the induced electrical stress. Adjusting these parameters helps researchers increase molecular entry while limiting cell damage. Because the technique can deliver DNA, RNA, proteins, and other molecules, optimization is needed to balance transfection efficiency with the ability of treated cells to recover.
Membrane resealing determines whether cells can recover after transient permeabilization. Under suitable recovery conditions, the pores close and the membrane returns toward its usual barrier function, allowing treated cells to remain useful for downstream experiments. If recovery is not adequately supported, the intended delivery benefit may be accompanied by greater cellular damage, reducing experimental utility.
The method supports intracellular delivery of plasmid DNA, RNA, proteins, and other molecules that normally cross cell membranes poorly. This range allows researchers to choose cargo according to the biological question, such as altering gene expression or supplying a protein. The same general approach can be applied to cultured cells, primary cells, and microorganisms.
Optimization centers on pulse strength, pulse duration, and the conditions provided after treatment. These factors influence both the extent of transient membrane permeabilization and the degree of cell damage. Researchers use the combination that produces sufficient molecular entry while preserving recovery, thereby improving transfection efficiency and the quality of subsequent cellular analyses.
Square Wave Electroporation is useful when researchers need to introduce nucleic acids, proteins, or other molecules into cells for functional studies. Its applications include investigating gene function, supporting genome-editing experiments, and engineering cells. Because it can be used with cultured cells, primary cells, and microorganisms, the technique fits a broad range of biological systems.