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          Institute: MPI für biophysikalische Chemie     Collection: Molekulare Biologie (Dr. Thomas M. Jovin)     Display Documents

ID: 233896.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
Complementary visualization of mitotic barley chromatin by field-emission scanning electron microscopy and scanning force microscopy
Authors:Schaper, A.; Roessle, M.; Formanek, H.; Jovin, T. M.; Wanner, G.
Date of Publication (YYYY-MM-DD):2000
Title of Journal:Journal of Structural Biology
Journal Abbrev.:J. Struct. Biol.
Start Page:17
End Page:29
Copyright:Jahrbuch 2001, Copyright MPG 2001
Review Status:not specified
Audience:Not Specified
Intended Educational Use:No
Abstract / Description: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.
Free Keywords:*Barley/cy [Cytology]
*Chromatin/ul [Ultrastructure]
Chromosomes/de [Drug Effects]
Comparative Study
Endopeptidase K/pd [Pharmacology]
Microscopy, Atomic Force
Microscopy, Electron, Scanning
Specimen Handling
Support, Non-U.S. Gov't
AFM; cell cycle; plant chromosome; DNA; scanning electron microscopy; scanning force microscopy; SFM
Last Change of the Resource (YYYY-MM-DD):2005-08-05
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:Botanical Institute of LMU–Munich, Menzingerstrasse 68, 80638 Munich, Germany
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