PUBLICATION

Embedding, serial sectioning and staining of zebrafish embryos using JB-4 resin

Authors
Sullivan-Brown, J., Bisher, M.E., and Burdine, R.D.
ID
ZDB-PUB-110110-41
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.
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.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping