Oral Intubation of Adult Zebrafish to Administer Experimental Compounds

Begin by loading a sterile oral intubation device with a suspension of nanostructured cytokine inclusion bodies  — the experimental nanoparticle.

Place a fasting anesthetized zebrafish onto a moist plastic tray.

Hold the fish steadily and gently open its mouth using the pipette tip attached to the oral intubation device.

Carefully insert the needle through the fish's mouth, down the esophagus.

Once the pipette tip passes the gills and encounters a slight resistance, avoid further insertion.

Gently inject the nanoparticles into the fish.

Upon injection, these cytokine nanoparticles come in direct contact with the epithelial cells that line the digestive tract of the zebrafish.

These cells internalize the nanoparticles through endocytosis.

Within the epithelial cell, the vesicle undergoes intracellular processing to release the cytokine nanoparticles.

These cytokine nanoparticles exit into the underlying connective tissue, interact with resident immune cells, and trigger intracellular signaling events, resulting in a cytokine-driven immune response.