Labelling cell structures and tracking cell lineage in zebrafish using SNAP-tag

Dev Dyn. 2011 Apr;240(4):820-7. doi: 10.1002/dvdy.22574. Epub 2011 Feb 28.

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acids / pharmacology
  • Animals
  • Animals, Genetically Modified
  • Cell Lineage* / genetics
  • Cell Tracking / methods*
  • Cellular Structures / cytology*
  • Cellular Structures / metabolism
  • Embryo, Nonmammalian / drug effects
  • Fixatives / pharmacology
  • Fluorescent Dyes / pharmacology*
  • Fluorescent Dyes / toxicity
  • Models, Biological
  • Staining and Labeling / methods*
  • Time-Lapse Imaging / methods
  • Zebrafish / embryology*
  • Zebrafish / genetics
  • Zebrafish / metabolism
  • Zebrafish / physiology

Substances

  • Acids
  • Fixatives
  • Fluorescent Dyes