Immunology microscopy of aldehyde-fixed yeast cells

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This chapter discusses the immunoelectron microscopy of aldehyde-fixed yeast cells. The immunolocalization of antigens in cells requires simultaneous preservation of structure and antigenicity. The difficulties of immunolocalization methods are centered on the methods used to preserve structure, which as a rule tend to degrade the ability of antigens (usually proteins) to be recognized by their cognate antibodies. The value of immunolocalization for cell biology was early recognized in the case of yeast at the level of the light microscope: immunolocalizations and, even more important, colocalizations using reagents labeled differentially with different fluorophores were developed and came into general use in the early 1980s. However, application of immunolocalization in electron microscopy of yeast lagged behind, largely because of the difficulty of adequately preserving both structure and antigenicity with the sample preparation methods then in general use. The chapter presents an overview and rationale for chemical fixation and processing of yeast cells for immuno-electron tomography (EM). The chapter presents the standard protocol for aldehyde fixation and LR white resin embedding and for obtaining and immunolabeling thin sections of yeast.

References (36)

  • M.N. Hall et al.

    Cell

    (1984)
  • P. Novick et al.

    Cell

    (1985)
  • R. Wright et al.
  • J. Pringle et al.
  • B. Byers et al.

    Methods Enzymol.

    (1991)
  • M. Clark

    Methods Enzymol.

    (1991)
  • B. Goud et al.

    Cell

    (1988)
  • M. Moeremans et al.

    J. Immunol. Methods

    (1984)
  • A.E. Adams et al.

    J. Cell Biol.

    (1984)
  • J. Kilmartin et al.

    J. Cell Biol.

    (1984)
  • E. van Tuinen et al.

    J. Histochem. Cytochem.

    (1987)
  • D. Preuss et al.

    Yeast

    (1991)
  • D. Preuss et al.

    Mol. Biol. Cell

    (1992)
  • J. Mulholland et al.

    J. Cell Biol.

    (1994)
  • J. Mulholland et al.

    Mol. Biol. Cell

    (1997)
  • J. Mulholland et al.

    Mol. Biol. Cell

    (1999)
  • K. McDonald et al.

    Methods Enzymol.

    (2002)
  • G. Griffiths

    Fine Structure Immunocytochemistry

    (1993)
  • View full text