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Archives of oto-rhino-laryngology

, Volume 233, Issue 1, pp 55–66 | Cite as

Immunofluorescence localization of proteins in semithin 0.2–1 μm frozen sections of the ear

A report of improved techniques including gelatin encapsulation and cryoultramicrotomy
  • Åke Flock
  • Yvonne Hoppe
  • Xu Wei
Article

Summary

The present work describes a high resolution technique for locating proteins in frozen sections of the inner ear by immunofluorescence. Dissected organs are encapsulated in gelatin, and sections 0.1–1 μm thick are cut at −100°C in a cryoultramicrotome. These are labelled with antibodies against two cytoskeletal proteins, actin and tubulin. Actin, which had previously only been described in the sensory cells, is found in the supporting cells as well. Tubulin is identified in the supporting cells and in outer spiral nerve fibres.

Schlüsselwörter

Innenohr Corti-Organ Aktin Tubulin Gefrierschnitte 

Key words

Inner ear Organ of Corti Actin Tubulin Frozen sections 

Immunofluoreszenzmikroskopische Lokalisation von Proteinen des Innenohres in 0.2–1 μm dicken Gefrierschnitten

Zusammenfassung

In dieser Arbeit wird eine Methode beschrieben, mit der man mit höherer Auflösung Proteine in Gefrierschnitten vom Innenohr durch Immunofluoreszens lokalisieren kann. Die Gewebestückchen werden in Gelatine eingebettet und Schnitte von 0,1–1 μm Dicke an einem Kryoultramikrotom bei −100° C geschnitten. Die Schnitte werden mit Antikörpern gegen die Proteine des Cytoskelets Aktin und Tubulin markiert. Actin, das bisher nur in der Sinneszelle lokalisiert war, wird nun auch in den Stützzellen gefunden. Tubulin wird in den Stützzellen und in den äußeren Spiralnervenfasern gefunden.

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Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • Åke Flock
    • 1
  • Yvonne Hoppe
    • 1
  • Xu Wei
    • 1
  1. 1.Dept. of PhysiologyKarolinska InstitutetStockholm 60Sweden

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