Multicolor fluorescent in situ mRNA hybridization (FISH) on whole mounts and sections.
Multicolor fluorescent in situ mRNA hybridization (FISH) on whole mounts and sections.
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DOI：
10.1007/978-1-61779-992-1_24
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Papalopulu, Nancy
中科院分区：
文献类型：
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作者：
Lea, Robert;Bonev, Boyan;Papalopulu, Nancy
原位杂交涉及反义 RNA 探针与 mRNA 转录物的杂交，它是表征组织、器官或整个生物体中基因表达的强大方法。作为整体安装 (WISH)，它可以在三个维度上检测 mRNA 转录本，同时与杂交之前或之后的切片相结合，它可以提供具有细胞分辨率的基因表达信息。 FISH 依赖于探针的荧光检测，是同时检测具有相似或重叠表达模式的转录本的首选方法，因为每个转录本都可以通过荧光团的选择来清楚地区分。在这里，我们描述了一种在爪蟾胚胎中进行多色 FISH 的方案，可进一步与抗体染色相结合。
In situ hybridization involves the hybridization of an antisense RNA probe to an mRNA transcript and it is a powerful method for the characterization of gene expression in tissues, organs, or whole organisms. Performed as a whole mount (WISH), it allows the detection of mRNA transcripts in three dimensions, while combined with sectioning, either before or after hybridization, it provides gene expression information with cellular resolution. FISH relies on the fluorescence detection of probes and is the method of choice for the simultaneous detection of transcripts with similar or overlapping expression patterns, as each can be clearly distinguished by the selection of fluorophore. Here, we describe a protocol for performing multicolor FISH in Xenopus embryos in whole mounts and sections that can be further combined with antibody staining.