荧光团
荧光
鬼臼苷
污渍
染色
生物物理学
光漂白
DNA
绿灯
亚历山福禄
化学
共焦显微镜
分子生物学
生物
细胞生物学
生物化学
细胞
光学
细胞骨架
植物
物理
红灯
遗传学
作者
Daniel Prieto,Gonzalo Aparicio,Matías Machado,Flavio R. Zolessi
摘要
Methyl green has long been known as a histological stain with a specific affinity for DNA, although its fluorescent properties have remained unexplored until recently. In this article, we illustrate the method for preparing a methyl green aqueous stock solution, that when diluted can be used as a very convenient fluorescent nuclear label for fixed cells and tissues. Easy procedures to label whole zebrafish and chick embryos are detailed, and examples of images obtained shown. Methyl green is maximally excited by red light, at 633 nm, and emits with a relatively sharp spectrum that peaks at 677 nm. It is very inexpensive, non-toxic, highly stable in solution and very resistant to photobleaching when bound to DNA. Its red emission allows for unaltered high resolution scanning confocal imaging of nuclei in thick specimens. Finally, this methyl green staining protocol is compatible with other cell staining procedures, such as antibody labeling, or actin filaments labeling with fluorophore-conjugated phalloidin.
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