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

Jessica Sullivan-Brown, Margaret E. Bisher, Rebecca D. Burdine

Research output: Contribution to journalArticlepeer-review

90 Scopus citations


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 resing-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 outlineg-from embryo preparation, embedding, sectioning and staining to visualizationg-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.

Original languageEnglish (US)
Pages (from-to)46-55
Number of pages10
JournalNature Protocols
Issue number1
StatePublished - Jan 2011

All Science Journal Classification (ASJC) codes

  • General Biochemistry, Genetics and Molecular Biology


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