Embedding, serial sectioning and staining of zebrafish embryos using JB-4 resin.
Publication Year
2011
Type
Journal Article
Abstract
Histological techniques are critical for observing tissue and cellular morphology. In this paper, we outline our protocol for embedding, serial sectioning, staining and visualizing zebrafish embryos embedded in JB-4 plastic resin-a glycol methacrylate-based medium that results in excellent preservation of tissue morphology. In addition, we describe our procedures for staining plastic sections with toluidine blue or hematoxylin and eosin, and show how to couple these stains with whole-mount RNA in situ hybridization. We also describe how to maintain and visualize immunofluorescence and EGFP signals in JB-4 resin. The protocol we outline-from embryo preparation, embedding, sectioning and staining to visualization-can be accomplished in 3 d. Overall, we reinforce that plastic embedding can provide higher resolution of cellular details and is a valuable tool for cellular and morphological studies in zebrafish.
Keywords
Journal
Nat Protoc
Volume
6
Issue
1
Pages
46-55
Date Published
01/2011
ISSN Number
1750-2799
Alternate Journal
Nat Protoc
PMID
21212782