Postembedding alpha-tubulin immunolabelling of isolated centrosomes

Histochem J. 1995 Mar;27(3):240-6.

Abstract

Accurate ultrastructural localization of the components of centrosomes is an important step toward the determination of their function. We have used an electron microscopy procedure to preserve centrosome-associated antigens which enables their high-resolution localization. The unique part of our procedure is the application of a post-sectioning fixation step which overcomes the poor section contrast and morphological appearance that limits the use of low-temperature processing and Lowicryl embedding. The efficacy of our approach is demonstrated by the efficient labelling of alpha-tubulin in the well-preserved and contrasted microtubule barrels of the centrides of isolated mammalian centrosomes.

MeSH terms

  • Centrosome / metabolism*
  • Centrosome / ultrastructure
  • Epoxy Resins
  • Humans
  • Immunohistochemistry
  • Lymphoid Tissue / metabolism
  • Microscopy, Electron
  • Microtubules / metabolism
  • Microtubules / ultrastructure
  • Plastic Embedding
  • Tubulin / metabolism*

Substances

  • Epoxy Resins
  • Tubulin