Encyclopedia of Experiments: Immunology
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Transcript
Take anesthetized dechorionated cystic fibrosis zebrafish embryos susceptible to Pseudomonas aeruginosa infection.
Inject fluorescent protein-expressing Pseudomonas aeruginosa into a major blood vessel, the duct of Cuvier.
Transfer injected embryos to a plate with media and a pigmentation-blocking agent to inhibit embryo pigmentation and visualize bacteria. Incubate.
Injected bacteria move through the circulation and invade organs.
In response, the host immune system gets activated, releasing pro-inflammatory mediators and recruiting immune cells.
Following incubation, inject a cocktail of virulent phages against Pseudomonas aeruginosa into the duct of Cuvier and transfer the embryos to a plate containing media and the pigmentation-blocking agent.
Incubate. Phages infect the bacteria and use host machinery to produce new phage particles.
Bacteria lyse, releasing phages that infect neighboring bacteria, resulting in a reduced pro-inflammatory response.
Using a fluorescent microscope, observe the reduced bacterial fluorescence in the zebrafish embryos over time, indicating phage effectiveness against Pseudomonas aeruginosa.
