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Lamellar body ultrastructure revisited: high-pressure freezing and cryo-electron microscopy of vitreous sections

Vanhecke, Dimitri ; Herrmann, Gudrun ; Graber, Werner ; Hillmann-Marti, Therese ; Mühlfeld, Christian ; Studer, Daniel ; Ochs, Matthias

In: Histochemistry and Cell Biology, 2010, vol. 134, no. 4, p. 319-326

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    Titre
    • Lamellar body ultrastructure revisited: high-pressure freezing and cryo-electron microscopy of vitreous sections
    Auteur
    • Vanhecke, Dimitri. Institute of Anatomy, University of Bern, Bern, Switzerland
    • Herrmann, Gudrun. Institute of Anatomy, University of Bern, Bern, Switzerland
    • Graber, Werner. Institute of Anatomy, University of Bern, Bern, Switzerland
    • Hillmann-Marti, Therese. Institute of Anatomy, University of Bern, Bern, Switzerland
    • Mühlfeld, Christian. Institute of Anatomy and Cell Biology, Justus-Liebig-University Giessen, Giessen, Germany
    • Studer, Daniel. Institute of Anatomy, University of Bern, Bern, Switzerland
    • Ochs, Matthias. Institute of Anatomy, University of Bern, Bern, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Histochemistry and Cell Biology, 2010, vol. 134, no. 4, p. 319-326. Springer-Verlag
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:314856
    Summary
    Lamellar bodies are the storage sites for lung surfactant within type II alveolar epithelial cells. The structure-function models of lamellar bodies are based on microscopic analyses of chemically fixed tissue. Despite available alternative fixation methods that are less prone to artifacts, such as cryofixation by high-pressure freezing, the nature of the lung, being mostly air filled, makes it difficult to take advantage of these improved methods. In this paper, we propose a new approach and show for the first time the ultrastructure of intracellular lamellar bodies based on cryo-electron microscopy of vitreous sections in the range of nanometer resolution. Thus, unspoiled by chemical fixation, dehydration and contrasting agents, a close to native structure is revealed. Our approach uses perfluorocarbon to substitute the air in the alveoli. Lung tissue was subsequently high-pressure frozen, cryosectioned and observed in a cryo-electron microscope. The lamellar bodies clearly show a tight lamellar morphology. The periodicity of these lamellae was 7.3nm. Lamellar bifurcations were observed in our cryosections. The technical approach described in this paper allows the examination of the native cellular ultrastructure of the surfactant system under near in vivo conditions, and therefore opens up prospectives for scrutinizing various theories of lamellar body biogenesis, exocytosis and recycling