High-Throughput Fluorescent Assay to Study Microbe-Worm Interactions

Begin with a tube containing C. elegans worms.

Transfer the worms to a multiwell plate with nutrient-rich agarose media containing either the control bacteria or the test bacteria.

During incubation, the worms ingest bacteria, which colonize the gut epithelium and release amino acids that serve as a nutrient source. These amino acids are metabolized into amino acid derivatives and stored in lysosome-related gut granules.

Next, add fresh buffer and transfer the worms to a black multiwell assay plate.

Add a chemical stress inducer to each well and seal the plate.

Place the plate in a fluorescence plate reader and record the fluorescence.

In worms, the stress inducer causes oxidative stress, which damages the gut epithelial cells.

This causes the lysosome-related gut granules to rupture, releasing amino acid derivatives, which, upon exposure to ultraviolet light, fluoresce blue.

Compare the fluorescence of worms exposed to the control and test bacteria to assess bacterial colonization efficiency and their impact on the host's oxidative stress response.

Adjust worm concentration to 15 worms per microliter in the 15 milliliter conical tube. Then, transfer 8 microliters of worm solution into each