A Technique to Develop a Parasite Infection in Intestinal Organoids via Microinjection

To microinject the parasites into the apical side of a 3D organoid, first, use a micropipette puller to prepare glass injection capillaries. Use forceps to cut the tip of the capillary to a 9- to 12-micrometer diameter, to enable an easy flow of sporozoites, or oocysts if you choose to inject oocysts directly, and use micro-loader tips to fill each capillary with a fast green dye-labeled oocyst or sporozoite suspension.

Then, load the first sporozoite-filled capillary into a microinjector, and use an inverted microscope at a 5x magnification and a constant pressure to microinject 100 to 200 nanoliters of the suspension into each organoid.