染色
斑马鱼
H&E染色
原位杂交
生物
病理
细胞生物学
信使核糖核酸
医学
生物化学
基因
作者
Jessica Sullivan-Brown,M. E. Bisher,Rebecca D. Burdine
标识
DOI:10.1038/nprot.2010.165
摘要
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.
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