1. Zebrafish Intestine Dissection
<ol>
	<li>After a specified period of time post intubation (e.g.,&#160;5 h and/or 24 h), place the fish using a net into 300 mg/L ethyl 3-aminobenzoate methanesulfonate (MS-222) solution for euthanasia. Make sure the operculum stops moving and there is no tail pinch reflex. Five minutes is normally sufficient.</li>
	<li>Pick up the euthanized animal with a net and place it on a filter paper. 
	NOTE: The filter paper is very useful for removing the adhesive tissue along the intestine.</li>
	<li>Using sharp dissection scissors, make a semi-circular incision from the anus to the operculum and open incision using fine tweezers. Cut both ends of the intestine, take out all the internal organs, and place them on the filter paper. 
	CAUTION: Work quickly to reduce cell metabolism and death. 
	NOTE: Alternatively, remove the adhesive tissue in PBS and on ice.</li>
	<li>Separate the intestine from internal organs making sure to keep its orientation (anterior to posterior intestinal segment) and stretch it out. Usually, the anterior segment of intestine is wider than posterior segment. Take care to obtain all of the intestine when dissecting. 
	NOTE: The posterior end is quite fine and fragile in small fish and may break off, particularly in animals &#60;0.7 g.</li>
	<li>Roll the intestine on the filter paper with tweezers in order to detach the adhesive tissue from the intestine.</li>
	<li>Proceed to prepare the intestine for various downstream analyses</li>
</ol>

<table><tbody><tr><td>MS-222&amp;nbsp;&amp;nbsp;</td><td>&amp;nbsp;Sigma Aldrich</td><td>E10521</td><td>powder</td></tr><tr><td>10x PBS&amp;nbsp;&amp;nbsp;</td><td>Sigma Aldrich</td><td>P5493</td><td></td></tr><tr><td>Filter paper&amp;nbsp;</td><td>&amp;nbsp;Filter-Lab</td><td>RM14034252</td><td></td></tr></tbody></table>

adult zebrafish intestine dissection: a technique to analyze compound absorption

Adult Zebrafish Intestine Dissection: A Technique to Analyze Compound Absorption

Source: Jie J. et. al., <a href="https://www.jove.com/v/58366/" target="_blank">Oral Intubation of Adult Zebrafish: A Model for Evaluating Intestinal Uptake of Bioactive Compounds</a>. J. Vis. Exp. (2018).
This video describes the technique of isolating the intestine to evaluate the intestinal uptake of bioactive compounds. In this sample, we will isolate the intestine from an adult zebrafish to analyze the uptake of immunostimulants.

Source: Jie J. et. al., Oral Intubation of Adult Zebrafish: A Model for Evaluating Intestinal Uptake of Bioactive Compounds. J. Vis. Exp. (2018).
This ...

- Place a euthanized fish on filter paper and pat it dry. Keep the fish on a dissecting mat. Pin the eye socket at the anterior end and the tail at the posterior end. Cut the skin and underlying muscle along the belly from the anal fin to the operculum, the hard covering over the gills.
Remove the operculum and pectoral fin. Cut the skin and underlying muscle from the gills posteriorly along the side of the fish to the upper anal fin. Now, open the incision with the help of a fine tweezer. Remove the gastrointestinal system from the body and place it on the filter paper.
Separate the intestine from other internal organs. The intestine is folded into three sections, the rostral intestinal bulb, mid intestine, and caudal intestine. Hold the anterior and posterior part of the intestine with the tweezer and gently stretch it out. Roll the intestine on the filter paper to remove all adhesive tissues. In the following protocol, we will isolate the intestine from an adult zebrafish to analyze the uptake of immuno stimulants.
- After euthanizing the fish according to the text protocol, place the fish on a piece of filter paper. Using sharp dissection scissors, make a semicircular incision from the anus to the operculum and open the incision with fine tweezers. Then cut both ends of the intestine, take out all of the internal organs, and place them on the filter paper. Next, separate the intestine from the internal organs and stretch it out, taking care to obtain all of the intestine. Finally, use tweezers to roll the intestine on the filter paper to detach the adhesive tissue.

adult-zebrafish-intestine-dissection-technique-to-analyze-compound

Research

JoVE Encyclopedia of Experiments

Biology

Danio rerio (zebrafish)

Adult

2.6K Views. - Place a euthanized fish on filter paper and pat it dry. Keep the fish on a dissecting 

Abstract

Protocol

<table><tbody><tr><td>MS-222&nbsp;&nbsp;</td><td>&nbsp;Sigma Aldrich</td><td>E10521</td><td>powder</td></tr><tr><td>10x PBS&nbsp;&nbsp;</td><td>Sigma Aldrich</td><td>P5493</td><td></td></tr><tr><td>Filter paper&nbsp;</td><td>&nbsp;Filter-Lab</td><td>RM14034252</td><td></td></tr></tbody></table>

Adult Zebrafish Intestine Dissection: A Technique to Analyze Compound Absorption

In This Article

Abstract

Protocol

Materials

Tags

Name	Company	Catalog Number	Comments
MS-222	Sigma Aldrich	E10521	powder
10x PBS	Sigma Aldrich	P5493
Filter paper	Filter-Lab	RM14034252