Gut Permeability Assay FD&C Method - C. elegans

Pick about 50 worms per condition and place them on a 35mm or 60mm plate

Prepare FD&C solution (10% stock) fresh - FD&C powder can be dissolved in water

Add the FD&C solution on to the plate containing the worms. The final concentration of the FD&C should be 2.5%

Leave the plate for about 3 hours  at room temperature

Remove the worms and teh FD&C dye into a 50 ml conical tube. Let the worms settle to the bottom (~ 5 minutes)

Remove the top FD&C dye (leave about 1ml in the tube. The worms should have settled to the bottom)

Add 30ml of water and dilute the FD&C.  Let the worms settle to the bottom (~ 5minutes)

Remove the excess liquid and add more water (~30ml) to wash the worms.  Let the worms settle to the bottom (~ 5minutes)

You should be able to visualize the worms at the bottom of the tube by now. Remove all the liquid and transfer the worms to a fresh plate with food.

Count worms that has dispersed blue dye.