Mechanical Disruption of Individual Worms to Assess Variability in Host-Bacteria Interactions

Begin with age-matched, sterile, freshly killed adult C. elegans worms with permeabilized cuticles, suspended in buffer.

These worms are infected with bacteria in their intestinal tracts.

Take a deep-well plate containing abrasive particles in buffer and transfer a single worm into each well.

Cover the plate with sealing film and a lid.

Incubate the plate at a low temperature to prevent overheating during tissue disruption.

Press the lid to seal the wells.

Using a tissue disruptor, mechanically shake the plate. The abrasive particles disrupt worm tissues, releasing intestinal bacteria.

Tap the plate and centrifuge it to pellet the contents.

Mix the contents to evenly distribute the bacteria, then transfer them to fresh wells.

Serially dilute the bacterial suspension in buffer.

Plate the dilutions onto agar plates and incubate to allow colony formation.

Count the colonies to quantify variation in intestinal bacterial loads across individual worms.

To prepare for mechanical disruption of worms, obtain a sterile two-milliliter deep well 96-well plate and a silicon plate cover.

Use a sterile scoop spatula to add a small amount of sterile 36 grid silicon carbide to each well such that the grid barel