FITC-Dextran Feeding: A Method to Quantify Intestinal Permeability in C. elegans
Published: April 30, 2023
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 |
Citer Cet Article
FITC-Dextran Feeding: A Method to Quantify Intestinal Permeability in C. elegans. J. Vis. Exp. (Pending Publication), e20167, doi: (2023).