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  Complementary visualization of mitotic barley chromatin by field-emission scanning electron microscopy and scanning force microscopy

Schaper, A., Roessle, M., Formanek, H., Jovin, T. M., & Wanner, G. (2000). Complementary visualization of mitotic barley chromatin by field-emission scanning electron microscopy and scanning force microscopy. Journal of Structural Biology, 129, 17-29.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F951-6 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0029-0CB3-B
Genre: Journal Article

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 Creators:
Schaper, A.1, Author              
Roessle, M., Author
Formanek, H., Author
Jovin, T. M.1, Author              
Wanner, G., Author
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: *Barley/cy [Cytology] *Chromatin/ul [Ultrastructure] Chromosomes/de [Drug Effects] Comparative Study Endopeptidase K/pd [Pharmacology] Microscopy, Atomic Force Microscopy, Electron, Scanning Mitosis Specimen Handling Support, Non-U.S. Gov't AFM; cell cycle; plant chromosome; DNA; scanning electron microscopy; scanning force microscopy; SFM
 Abstract: The surface structure of mitotic barley chromatin was studied by field-emission scanning electron microscopy (FESEM) and scanning force microscopy (SFM). Different stages of the cell cycle were accessible after a cell suspension was dropped onto a glass surface, chemical fixed, and critically point dried. Imaging was carried out with metal-coated specimen or uncoated specimen (only for SFM). The spatial contour of the chromatin could be resolved by SFM correlating to FESEM data. The experimentally determined volume of the residue chromatin during mitosis was within the range of 65-85 microm(3). A comparison with the theoretically calculated volume indicated a contribution of about 40% of internal cavities. Decondensation of chromosomes by proteinase K led to a drastic decrease in the chromosome volume, and a 3-D netlike architecture of the residue nucleoprotein material, similar to that in the intact chromosome, was obvious. Incubation of metaphase chromosomes in citrate buffer permitted access to different levels of chromatin packing. We imaged intact chromosomes in liquid by SFM without any intermediate drying step. A granular surface was obvious but with an appreciably lower resolution. Under similar imaging conditions proteinase K-treated chromosomes exhibited low topographic contrast but were susceptible to plastic deformations.

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Language(s): eng - English
 Dates: 2005-08-052000
 Publication Status: Published in print
 Pages: -
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 Identifiers: eDoc: 233896
Other: 23129
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Title: Journal of Structural Biology
  Alternative Title : J. Struct. Biol.
Source Genre: Journal
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Pages: - Volume / Issue: 129 Sequence Number: - Start / End Page: 17 - 29 Identifier: -