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Gut Permeability Assay FD&C Method - C. elegans
Author: Harshi Peiris, updated date: , view: 21, Q&A: 0
Tags: Worm and Gut

Procedure

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.


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