Golgi twins in mitosis revealed by genetically encoded tags for live cell imaging and correlated electron microscopy

  • Ben N. G. Giepmans
  • Guido M. Gaietta
  • Thomas J. Deerinck
  • Stephen R. Adams
  • Roger Y. Tsien
  • Mark H. Ellisman
Conference paper

Abstract

The dynamic behaviour of cells is a consequence of the coordinated and elaborate interactions between complexes of macromolecules that constitute their formed structures or organelles. Our understanding of basic cell structure and function has been greatly aided by the identification of proteins at the ultrastructural level. Advances in molecular biology, organic chemistry, and materials science have recently led to the creation of several new classes of fluorescent probes for live cell imaging and correlated light and electron microscopy (EM; Fig. 1; [1,2]). Here we show the application of (i) small organic fluorescent dyes, (ii) nanocrystals (“quantum dots”), (iii) fluorescent proteins, including the optimized mCherry, and (iv) genetic encoded tetracysteine tags complexed with biarsenical dyes (FlAsH and ReAsH), to determine protein dynamics and ultrastructural localization of proteins and organelle structures.

Keywords

Golgi apparatus mitosis correlated microscopy 

References

  1. 1.
    Giepmans BNG, Adams SR, Ellisman MH & Tsien RY (2006). The fluorescent toolbox for assessing protein location and function. Science 312, 217–224.CrossRefADSGoogle Scholar
  2. 2.
    Sosinsky GE, Giepmans BNG, Deerinck TJ, Gaietta GM & Ellisman MH (2007). Markers for correlated light and electron microscopy. In: Cellular Electron Microscopy, ed. McIntosh JR, San Diego: Academic Press, Elsevier Inc. 573–589.Google Scholar
  3. 3.
    Gaietta, GM, Giepmans BN, Deerinck TJ, Smith WB, Ngan L, Llopes J, Adams SR, Tsien RY & Ellisman MH (2006). Golgi twins in late mitosis revealed by genetically encoded tags for live cell imaging and correlated electron microscopy. PNAS 103, 17777–82.CrossRefADSGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2008

Authors and Affiliations

  • Ben N. G. Giepmans
    • 1
    • 2
    • 4
  • Guido M. Gaietta
    • 1
  • Thomas J. Deerinck
    • 1
  • Stephen R. Adams
    • 2
  • Roger Y. Tsien
    • 2
    • 3
  • Mark H. Ellisman
    • 1
  1. 1.Center for Research in Biological Systems and Dept. of NeurosciencesNational Center for Microscopy and Imaging ResearchUSA
  2. 2.Dept. of PharmacologyUniversity of CaliforniaSan Diego; La JollaUSA
  3. 3.Howard Hughes Medical InstituteUniversity of CaliforniaSan Diego; La JollaUSA
  4. 4.Molecular Imaging & EM, Dept of Cell BiologyUniversity Medical Center GroningenGroningenThe Netherlands

Personalised recommendations