DOI: 10.1101/493536Dec 13, 2018Paper

Dissection of intestines from larval zebrafish for molecular analysis

BioRxiv : the Preprint Server for Biology
Bilge SanLeonie Kamminga

Abstract

Epigenetic data obtained from whole zebrafish embryos or larvae may mask or dilute organ-specific information. Fluorescence activated cell sorting can diverge cells from their native state, and cryosections often yield insufficient material for molecular analysis. Here, we present a reproducible method for larval intestinal isolation at 5, 7, and 9 days post-fertilization, using the intestine-specific transgene tgBAC(cldn15la:GFP). With tweezers, the intestine can be pulled out of the abdomen in one smooth motion. Upon removal of adhering tissues, intestines can be directly used for analyses. Each dissection takes 3-6 minutes per fish. We demonstrate that 10 and 25 dissected intestines yield enough material for RNA-sequencing and ChIP-sequencing, respectively. This method results in high quality, live material, suitable for many downstream applications.

Related Concepts

Abdomen
Tissue Dissection
Embryo
Fertility
Intestines
Larva
Zebrafish
Cryoultramicrotomy
Organ
Isolation Aspects

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