Contour-length fluctuation of polymers: NMR experiments and statistical physics

1973 ◽

Vol 31 ◽

pp. 596-597

Author(s):

C. N. Gordon

Keyword(s):

Electron Microscopy ◽

Aqueous Salt Solution ◽

Contour Length ◽

Cleavage Surface ◽

Mica Substrate ◽

Electron Microscopic ◽

Replica Technique ◽

Circular Dna ◽

Transmission Electron ◽

Electron Microscopic Technique

Gordon and Kleinschmidt have described a new preparative technique for visualizing DNA by electron microscopy. This procedure, which is a modification of Hall's “mica substrate technique”, consists of the following steps: (a) K+ ions on the cleavage surface of native mica are exchanged for Al3+ ions by ion exchange. (b) The mica, with Al3+ in the exchange sites on the surface, is placed in a dilute aqueous salt solution of DNA for several minutes; during this period DNA becomes adsorbed on the surface. (c) The mica with adsorbed DNA is removed from the DNA solution, rinsed, dried and visualized for transmission electron microscopy by Hall's platinum pre-shadow replica technique.In previous studies of circular DNA by this technique, most of the molecules seen were either broken to linears or extensively tangled; in general, it was not possible to obtain suitably large samples of open extended molecules for contour length measurements.

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Existence of DNA in yeast microbodies

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100082029 ◽

1978 ◽

Vol 36 (2) ◽

pp. 232-233

Author(s):

Masako Osumi ◽

Misuzu Nagano ◽

Hiroko Kazama

Keyword(s):

Catalase Activity ◽

Buoyant Density ◽

Model System ◽

Contour Length ◽

Native State ◽

Normal Alkane ◽

Remarkable Increase ◽

Dna Molecule ◽

Mitochondrial Dnas

We have found that microbodies appeared profusely together with a remarkable increase in catalase activity in normal alkane-grown cells of hydrocarbon-utilizing Candida yeasts, and that the microbodies multiplied by division in these cells. These features of Candida yeasts seem to provide a useful model system for studies on the biogenesis of the microbody. Subsequently, we have succeeded in isolation of Candida microbodies in an apparently native state, as judged biochemically and morphologically. The presence of DNA in the purified microbody fraction thus obtained was proved by the diphenylamine method. DNA molecule of about 15 urn in contour length was released from an isolated microbody. The physicochemical analyses of the microbody DNA revealed that its buoyant density differed from nuclear and mitochondrial DNAs. All these results lead us to the possibility that there is a novel type of DNA in microbodies.

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Nucleic Acid Molecules Prepared by Monolayer Techniques

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094140 ◽

1972 ◽

Vol 30 ◽

pp. 178-179

Author(s):

Dimitrij Lang

Keyword(s):

Electron Microscopy ◽

Standard Deviation ◽

Nucleic Acid ◽

Cytochrome C ◽

Nucleic Acids ◽

Contour Length ◽

Sample Standard Deviation ◽

Molecular Weights ◽

Length Measurements ◽

Protein Monolayer

The success of the protein monolayer technique for electron microscopy of individual DNA molecules is based on the prevention of aggregation and orientation of the molecules during drying on specimen grids. DNA adsorbs first to a surface-denatured, insoluble cytochrome c monolayer which is then transferred to grids, without major distortion, by touching. Fig. 1 shows three basic procedures which, modified or not, permit the study of various important properties of nucleic acids, either in concert with other methods or exclusively:1) Molecular weights relative to DNA standards as well as number distributions of molecular weights can be obtained from contour length measurements with a sample standard deviation between 1 and 4%.

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Quantification of cell-surface expressed antigenic sites with 5-nm gold markers: Getting close to biologically relevant data

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100157231 ◽

1989 ◽

Vol 47 ◽

pp. 1050-1051

Author(s):

Etienne de Harven ◽

Hilary Christensen ◽

Richard Leung ◽

Cameron Ackerley

Keyword(s):

Cell Surface ◽

Human Peripheral Blood ◽

Contour Length ◽

Thin Sections ◽

Gold Particles ◽

Biologically Relevant ◽

Transmission Electron ◽

Entire Cell ◽

Electron Imaging ◽

Cell Contour

The T-derived subset of human peripheral blood normal lymphocytes has been selected as a model system to study the usefulness of 5 nm gold markers for quantification of single epitopes expressed on cell surfaces. The chosen epitopes are parts of the CD3 and CD5 molecules and can be specifically identified by hybridoma produced monoclonal antibodies (MoAbs; LEU-4 and LEU-1; Becton-Dick- inson, Mountain view, CA) . An indirect immunolabeling procedure, with goat anti-murine IgG adsorbed on the surface of 5 nm colloidal gold particles (GAM-G5, Janssen Pharmaceutica, Beerse, Belgium) has been used. Backscattered Electron Imaging (BEI) in a field emission scanning electronmicroscope (SEM) and transmission electron microscopy of thin sections of lymphocytes labeled before plastic embedding, were both used to identify and quantitate gold labeled cell surface sites, Estimating that the thickness of “silver” sections is approximately 60 nm and counting the number of gold particles on the entire cell perimeter, we calculated that, for LEU-4, the number of markers per um2 of cell surface is in the 140-160 range (Fig.l). Cell contour length measurements indicated that the surface of one lymphocyte is approximately 130-160 um2 that of a smooth sphere of identical diameter, reflecting the role of microvilli in expanding the surface area. The total number of gold labeled sites on the surface of one lymphocyte averages, therefore between 20,000 and 24,000 per cell.

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