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Labelling cell structures and tracking cell lineage in zebrafish using SNAP‐tag
Author(s) -
Campos Cláudia,
Kamiya Mako,
Banala Sambashiva,
Johnsson Kai,
GonzálezGaitán Marcos
Publication year - 2011
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.22574
Subject(s) - biology , zebrafish , snap , lineage (genetic) , microbiology and biotechnology , cell , cell lineage , labelling , computational biology , genetics , gene , cellular differentiation , biochemistry , computer science , computer graphics (images)
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. © 2011 Wiley‐Liss, Inc.