Labelling Cell Structures and Tracking Cell Lineage in Zebrafish Using SNAP-Tag
Labelling Cell Structures and Tracking Cell Lineage in Zebrafish Using SNAP-Tag
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DOI：
10.1002/dvdy.22574
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发表时间：
2011-04-01
影响因子：
2.5
通讯作者：
Gonzalez-Gaitan, Marcos
中科院分区：
文献类型：
--
作者：
Campos, Claudia;Kamiya, Mako;Gonzalez-Gaitan, Marcos
我们提出了一种用合成荧光团特异性标记斑马鱼融合蛋白的方法。该方法使用SNAP-标签技术和各种合成荧光团的O-6-苄基鸟嘌呤衍生物。我们演示了如何使用该方法来标记斑马鱼的亚细胞结构，如细胞核，细胞膜和内体膜。合成荧光团的稳定性使它们成为长期成像的有吸引力的选择，并且与大多数自体荧光蛋白不同，允许使用酸性固定剂，如三氯乙酸。此外，使用O-6-苄基鸟嘌呤衍生物携带笼状荧光素允许细胞谱系跟踪通过光脱保护的荧光团和其检测通过荧光显微镜或通过免疫组织化学固定后，使用抗荧光素抗体。Developmental Dynamics 240：820-827，2011. (C)2011 Wiley-Liss，Inc.
We present a method for the specific labelling of fusion proteins with synthetic fluorophores in Zebrafish. The method uses the SNAP-tag technology and O-6-benzylguanine derivatives of various synthetic fluorophores. We demonstrate how the method can be used to label subcellular structures in Zebrafish such as the nucleus, cell membranes, and endosomal membranes. The stability of the synthetic fluorophores makes them attractive choices for long-term imaging and allows, unlike most of the autofluorescent proteins, the use of acid fixatives such as trichloroacetic acid. Furthermore, the use of O-6-benzylguanine derivatives bearing caged fluorescein allows cell lineage tracing through photo-deprotection of the fluorophore and its detection either through fluorescence microscopy or through immunohistochemistry after fixation using anti-fluorescein antibodies. Developmental Dynamics 240:820-827, 2011. (C) 2011 Wiley-Liss, Inc.