title: Labelling cell structures and tracking cell lineage in zebrafish using SNAP-tag.
abstract: 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.
authors.affiliation: Departement de Biochimie, Université de Genève, Genève, Switzerland.
authors.forename: Cláudia
authors.initials: C
authors.lastname: Campos
authors.forename: Mako
authors.initials: M
authors.lastname: Kamiya
authors.forename: Sambashiva
authors.initials: S
authors.lastname: Banala
authors.forename: Kai
authors.initials: K
authors.lastname: Johnsson
authors.forename: Marcos
authors.initials: M
authors.lastname: González-Gaitán
mesh_terms: D000143:Acids / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D019070:Cell Lineage / Q000235:genetics; D058948:Cell Tracking / Q000379:methods*; D022082:Cellular Structures / Q000166:cytology* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000187:drug effects; D005404:Fixatives / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology* / Q000633:toxicity; D008954:Models, Biological; D013194:Staining and Labeling / Q000379:methods*; D059008:Time-Lapse Imaging / Q000379:methods; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000378:metabolism / Q000502:physiology
chemical_list: D000143:Acids; D005404:Fixatives; D005456:Fluorescent Dyes