FITC-Dextran Feeding: A Method to Quantify Intestinal Permeability in C. elegans

Abstract

Source: Le, T. A. N., et al. Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans. J. Vis. Exp. (2019).

This video introduces a method of to visualize and measure the integrity of the C. elegans intestine lumen by feeding worms FITC-labeled dextran.  The example protocol shows the assay done with 3,3’-diindolylmethane (DIM)-treated and pathogen-fed worms.

Protocol

This protocol is an excerpt from Le et al, Measuring the Effects of Bacteria and Chemicals on the Intestinal Permeability of Caenorhabditis elegans, J. Vis. Exp. (2019). Preparation of NGM Plates for Testing the Effects of a Chemical (DIM)…

Representative Results

Figure 1: The effect of DIM on the intestinal permeability of C. elegans fed P. aeruginosa. Microscopy images of worms from (A) E. coli OP50 without FITC-dextran feeding, (B) E. coli with FITC-dextran feeding, (C</str…

Materials

3,3’-diindolylmethane	Sigma	D9568
90×15 mm Petri dishes	SPL Life Sciences, South Korea	10090
Bactor Agar	Beckton Dickinson	REF. 214010
Formaldehyde solution	Sigma	F1635
Caenorhabditis elegans N2	Caenorhabditis Genetics Center (CGC)		Wild type
Cholesterol	Sigma	C3045
Costa Assay Plate, 96 Well Black With Clear Flat Bottom Non-treated, No Lid Polystyrene	Corning Incorporated	REF. 3631
Dimethyl sulfoxide	Sigma	D2650
Escherichia coli OP50	Caenorhabditis Genetics Center (CGC)
Fluorescein isothiocyanate – dextran	Sigma	FD10S
Harmony software	PerkinElmer		verson 3.5
Magnesium sulfate heptahydrate	Fisher Bioreagents	BP2213-1
Fluoromount aqueous mounting medium	Sigma	F4680
Operetta CLS High-Content Analysis System	PerkinElmer	HH16000000
Peptone	Merck	EMD 1.07213.1000
Pseudomonas aeruginosa PA01	Korean Collection for Type Culture	KCTC NO. 1637
Sodium Chloride	Fisher Bioreagents	BP358-1
Stereo Microscope	Nikon, Japan	SMZ800N