A Robust Procedure for Distinctively Visualizing Zebrafish Retinal Cell Nuclei Under Bright Field Light Microscopy
A Robust Procedure for Distinctively Visualizing Zebrafish Retinal Cell Nuclei Under Bright Field Light Microscopy
复制标题
DOI：
10.1369/0022155412471535
复制
发表时间：
2013-03-01
影响因子：
3.2
通讯作者：
Wei, Xiangyun
中科院分区：
文献类型：
--
作者：
Fu, Jinling;Fang, Wei;Wei, Xiangyun
为了在明亮的视野显微镜下同时观察斑马鱼视网膜的单个细胞核和组织形态，有必要建立一种在薄组织切片中特异和灵敏地对细胞核进行染色的程序。这种必要性源于视网膜的高核密度和视锥感光细胞高度去凝集的染色质，这显著地减少了它们的核信号，使核很难与可能的高细胞质背景染色区分开来。在这里，我们优化了一种整合了JB4塑料包埋和Feulgen反应的方法，用于在强场光显微镜下显示斑马鱼视网膜细胞核。这种方法产生了高度特异的核染色，胞浆背景最少，使我们能够区分单个视网膜细胞核，尽管它们包装紧密。核染色也足够敏感，可以区分斑马鱼锥体细胞核中的常染色质和异染色质。此外，该方法可以与原位杂交相结合，同时显示细胞核和mRNA的表达模式。由于其极高的特异性和敏感性，该方法可以扩展到量化细胞密度和分析整个视网膜或其他组织的整体染色质组织。(组织化学细胞化学杂志61：248-256,2013)
To simultaneously visualize individual cell nuclei and tissue morphologies of the zebrafish retina under bright field light microscopy, it is necessary to establish a procedure that specifically and sensitively stains the cell nuclei in thin tissue sections. This necessity arises from the high nuclear density of the retina and the highly decondensed chromatin of the cone photoreceptors, which significantly reduces their nuclear signals and makes nuclei difficult to distinguish from possible high cytoplasmic background staining. Here we optimized a procedure that integrates JB4 plastic embedding and Feulgen reaction for visualizing zebrafish retinal cell nuclei under bright field light microscopy. This method produced highly specific nuclear staining with minimal cytoplasmic background, allowing us to distinguish individual retinal nuclei despite their tight packaging. The nuclear staining is also sensitive enough to distinguish the euchromatin from heterochromatin in the zebrafish cone nuclei. In addition, this method could be combined with in situ hybridization to simultaneously visualize the cell nuclei and mRNA expression patterns. With its superb specificity and sensitivity, this method may be extended to quantify cell density and analyze global chromatin organization throughout the retina or other tissues. (J Histochem Cytochem 61:248-256, 2013)