Protocol

Animal Cell Electroporation and Electrofusion Protocols

Volume 48 of the series Methods in Molecular Biology pp 185-197

Electrotransfection with “Intracellular” Buffer

  • Maurice J. B. van den HoffAffiliated withDepartment of Anatomy and Embryology, Academic Medical Center, University of Amsterdam
  • , Vincent M. ChristoffelsAffiliated withDepartment of Anatomy and Embryology, Academic Medical Center, University of Amsterdam
  • , Wil T. LabruyèreAffiliated withDepartment of Anatomy and Embryology, Academic Medical Center, University of Amsterdam
  • , Antoon F. M. MoormanAffiliated withDepartment of Anatomy and Embryology, Academic Medical Center, University of Amsterdam
  • , Wouter H. LamersAffiliated withDepartment of Anatomy and Embryology, Academic Medical Center, University of Amsterdam

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Abstract

Introduction of foreign molecules, such as DNA, RNA, or proteins, into living cells is a powerful means to test the biological functions of these molecules. One of the techniques by which foreign molecules can be introduced into living cells is electroporation (for reviews, see refs. 1,2). Compared to other techniques for the introduction of molecules into living cells, electroporation does not seem to cause prolonged alterations in the biological structure and function of the target cells. Nevertheless, the pores that are induced by the electroporation technique (35) can be visualized by rapid-freezing electron microscopy techniques (6) and allow a direct contact between the cytoplasm of the cells and the electroporation medium. The components of the electroporation medium can, therefore, penetrate into the cell interior as long as the pores exist (7). In this perspective, it is obvious that cells are exquisitely sensitive to the composition of the electroporation medium (810) and that an electroporation medium that resembles the composition of the cytoplasm would probably enhance the viability of the cells after electroporation.