ZFIN ID: ZDB-PUB-110110-41
Embedding, serial sectioning and staining of zebrafish embryos using JB-4 resin
Sullivan-Brown, J., Bisher, M.E., and Burdine, R.D.
Date: 2011
Source: Nature Protocols   6(1): 46-55 (Journal)
Registered Authors: Burdine, Rebecca, Sullivan-Brown, Jessica
Keywords: Cell biology, Imaging, Model organisms, Imaging, Developmental biology
MeSH Terms:
  • Animals
  • Embryo, Nonmammalian/anatomy & histology*
  • Embryo, Nonmammalian/chemistry
  • Embryo, Nonmammalian/ultrastructure
  • Green Fluorescent Proteins/analysis
  • Microscopy, Fluorescence
  • Microtomy/methods*
  • Plastic Embedding/methods*
  • Polymers
  • Staining and Labeling/methods
  • Zebrafish/embryology*
PubMed: 21212782 Full text @ Nat. Protoc.
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.