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Nucleic Acids Research, 1980, Vol. 8, No. 2 265-278
© 1980

Article

Chromatin freeze fracture electron microscopy: a comparative study of core particles, chromatin, metaphase chromosomes, and nuclei

Jean Lepault1,*, Stanley Bram1,+, Jacques Escaig2 and Wayne Wray3

1Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15 2Laboratoire de Microscopie Electronique Appliquéa la Biologie, Université de Paris IV, France 3Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030, USA

Received October 18, 1979.

Chromatin gels, metaphase chromosomes, and intact nuclei were studied by freeze fracturing followed by electron microscopy. The results complement and extend those obtained by classical electron microscopy techniques as they are obtained without fixation or dehydration. The freeze fracturing technique permits a determination of the hydrated diameters of nucleosanes in chromatin and in nuclei to be 13 nm by comparing to simultaneously studied test objects. Nucleosomes in chromatin fibers are closely spaced but are discrete particles in all conditions studied. In the presence of divalent ions, most chromatin in solution, chromosomes, and nuclei is organized into fibers whose thickness is larger than 40 nm. The images are not at all compatible with a super bead organization of the nucleofilament. Freeze fractures of intact nuclei provides information on the distribution of chromatin in a hydrated unfixed state. The images suggest that most of the chromatin is localized in large domains in contact with the inner nuclear membrane.

*E.M.B.O., Electron Microscropy Unit, Heidelburg, West Germany.

+Institut Ourie, 11 rue Pierre et Marie Ourie, Paris 5, France.


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