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Cell Staining: Fluorescent Labelling of the Golgi Apparatus

  1. Selma Y Dejgaard,
  2. Kurt Dejgaard,
  3. John F Presley

Published Online: 15 DEC 2010

DOI: 10.1002/9780470015902.a0002633.pub2

eLS

eLS

How to Cite

Dejgaard, S. Y., Dejgaard, K. and Presley, J. F. 2010. Cell Staining: Fluorescent Labelling of the Golgi Apparatus. eLS. .

Author Information

  1. McGill University, Montreal, Quebec, Canada

Publication History

  1. Published Online: 15 DEC 2010

This is not the most recent version of the article. View current version (14 MAY 2015)

Abstract

Immunofluorescence has been the primary method for labelling the Golgi apparatus for light microscopic observation in fixed cells. The Golgi apparatus was initially labelled in living cells by using that the fluorescent ceramide analogue N-6[7-nitro-2,1,3-benzoxadiazol-4-yl] aminocaproyl sphingosine or by microinjection of fluorescently conjugated antibodies to exposed Golgi epitopes. With the common availability of the green fluorescent protein (GFP) and advanced fluorescence microscopes including confocal microscopes, imaging of the Golgi apparatus in living cells is now a major experimental approach. Time-lapse imaging of living cells has also been combined with photobleach techniques in order to study the dynamics of transient protein association with the Golgi apparatus. Similar approaches have been used to study the dynamics of the cargo transport through the exocytic pathway.

Key Concepts:

  • Immunofluorescence can be used to visualise proteins in the Golgi apparatus or its subdomains.

  • Live-cell visualisation of the Golgi apparatus is possible with GFP-tagged proteins.

  • Photobleaching and photoactivation techniques permit visualisation of protein dynamics and flux through the Golgi apparatus in living cells.

Keywords:

  • Golgi apparatus;
  • exocytosis;
  • immunofluorescence;
  • vital staining;
  • GFP;
  • photobleach;
  • fluorescence microscopy