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

Author(s): Sullivan-Brown, Jessica; Bisher, Margaret E; Burdine, Rebecca D

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Abstract: Histological techniques are critical for observing tissue and cellular morphology. Here, 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 (H&E), and show how to couple these stains with whole-mount RNA in situ hybridization. We also describe how to maintain and visualize immunofluorescence and GFP signals in JB-4™ resin. The protocol we outline – from embryo preparation, embedding, sectioning and staining to visualization - can be accomplished in three days. 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.
Publication Date: Jan-2011
Electronic Publication Date: 16-Dec-2010
Citation: Sullivan-Brown, Jessica, Bisher, Margaret E, Burdine, Rebecca D. (2011). Embedding, serial sectioning and staining of zebrafish embryos using JB-4 resin. Nature Protocols, 6 (1), 46 - 55. doi:10.1038/nprot.2010.165
DOI: doi:10.1038/nprot.2010.165
ISSN: 1754-2189
EISSN: 1750-2799
Pages: 46 - 55
Type of Material: Journal Article
Journal/Proceeding Title: Nature Protocols
Version: Final published version. Article is made available in OAR by the publisher's permission or policy.

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