JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:22 min • October 30th, 2025
Begin with germ-free adult C. elegans worms suspended in media.
Introduce the test bacterium and incubate.
As the worms feed, bacteria enter the intestinal lumen and adhere to the epithelium.
Centrifuge to pellet the worms and remove the non-ingested bacteria.
Resuspend in buffer, centrifuge, and remove the remaining non-ingested bacteria.
Add media containing heat-killed E. coli and incubate.
During feeding, the heat-killed E. coli displaces and expels non-adhered test bacteria from the intestinal lumen.
Centrifuge and remove the supernatant containing the expelled bacteria.
Wash with buffer, centrifuge, and remove the supernatant.
Chill the worms on ice to induce paralysis, preventing further ingestion.
Add a bleach solution and incubate to kill residual external bacteria.
Remove the bleach and add cold buffer.
Centrifuge and remove the supernatant to eliminate residual bleach.
The worms now retain only viable test bacteria adhered to the intestinal epithelium, enabling quantification of colonization.
Begin by resuspending the worm sample in one milliliter of M9TX-01 in a microcentrifuge tube. Collect the adult worms by centrifugation and discard the supernatant. Using the same centrifugation
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