Amoeba Electroporation
Amoeba Electroporation - Related Videos
02:04
Transfection of Wild-Type Amoeba via Electroporation with Bacterial Feeding Support
0 Views •
2025
Source: Paschke, P., et al. Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria. J. Vis. Exp. (2019).This video demonstrates the procedure for introducing a plasmid carrying a target gene into amoeba cells via electroporation, followed by recovery in a bacterial supplement to enable efficient gene delivery in wild-type amoeba.
04:15
Detecting Bacterial Virulence Using Amoeba as a Host System
0 Views •
2026
Source: Dubois, V., et al. Identification of Virulence Markers of Mycobacterium abscessus for Intracellular Replication in Phagocytes. J. Vis. Exp. (2018)This video demonstrates the procedure of co-culturing amoebae with mutant bacterial strains to evaluate their intracellular survival and identify virulence markers through differences in colony counts.
03:25
Amoeba-Based Enrichment of Giant Viruses from an Environmental Water Sample
0 Views •
2026
Source: Bou Khalil, J. Y.,et al. A Rapid Strategy for the Isolation of New Faustoviruses from Environmental Samples Using Vermamoeba vermiformis. J. Vis. Exp. (2016)This video demonstrates the enrichment of giant viruses from an environmental water sample using amoeba-based co-culture to enable downstream detection and analysis.
04:12
Isolation of a Single Giant Virus from a Mixed Viral Population Using Amoebae
0 Views •
2026
Source: Sahmi-Bounsiar, D. et al. Single Cell Micro-aspiration as an Alternative Strategy to Fluorescence-activated Cell Sorting for Giant Virus Mixture Separation. J. Vis. Exp. (2019)This video demonstrates single-cell micro-aspiration to isolate and expand a single giant virus population from a mixed viral population using amoebae as host cells.
11:57
Electroporation of Mycobacteria
0 Views •
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
During the pulse, the electrical field briefly changes plasma-membrane permeability, allowing extracellular DNA, RNA, or other selected molecules to cross. Once the field stops, the pores close as the membrane reseals, so delivery depends on the timing of transport relative to pore formation and closure. This transient behavior enables intracellular cargo delivery without requiring permanent membrane disruption.
Pulse strength and duration must be balanced rather than maximized. Stronger or longer exposure can interfere with membrane integrity and reduce recovery, whereas insufficient electrical treatment may limit molecular entry. Cell condition also changes the outcome, making these variables central to optimization. Researchers therefore evaluate delivery together with post-pulse survival and recovery under suitable culture conditions.
Membrane resealing does not end the experiment; cells must recover under suitable culture conditions so researchers can assess delivery and downstream effects. Poor recovery can obscure whether a weak result reflects inadequate molecular entry or reduced cellular fitness after treatment. Monitoring recovery therefore helps researchers interpret transient expression and functional assays more reliably.
A typical workflow prepares amoeba cells, combines them with the intended DNA, RNA, or other molecule, applies a brief electrical pulse, and then returns the cells to suitable culture conditions. Later analysis determines whether the delivered material produced the intended effect, such as reporter activity or altered gene function. This separates delivery, recovery, and biological interpretation.
Optimization focuses on pulse strength, pulse duration, cell condition, and recovery conditions because each can influence electroporation efficiency. The useful setting is not simply the one that produces the strongest electrical treatment. Researchers seek conditions that support molecular entry while allowing cells to recover sufficiently for subsequent expression measurements or functional studies.
The technique supports transient gene expression, reporter assays, and gene-function analysis. Delivered nucleic acids can be used to examine whether a gene or regulatory construct produces a measurable cellular effect without requiring the experiment to focus only on long-term genetic changes. These applications make the method useful for testing molecular function and optimizing delivery conditions in amoebae.
By enabling functional manipulation in amoebae, the approach can help investigate cell motility, phagocytosis, signaling, and host-microbe interactions. Researchers can connect delivered genetic material with changes in these biological processes, using the resulting cellular responses to study gene function and the mechanisms underlying amoeba behavior in relevant experimental systems.