scholarly journals FIBRILLAR DIFFERENTIATION IN MYXOMYCETE PLASMODIA

1965 ◽  
Vol 25 (2) ◽  
pp. 305-318 ◽  
Author(s):  
Sister M. A. McManus ◽  
L. E. Roth
Keyword(s):  
Electron Microscope ◽  
Molecular Species ◽  
Physarum Polycephalum ◽  
Mitotic Apparatus ◽  
Thin Sections ◽  
Fixation Methods ◽  
Different Types ◽  
Protoplasmic Movement

Myxomycete plasmodia of four different types (not including Physarum polycephalum) were studied in thin sections viewed in the electron microscope. In the cytoplasm of the protoplasmodia of Clastoderma debaryanum and the phaneroplasmodia of Fuligo septica fixed in situ, fibrillar differentiations of three rather distinct kinds were observed. One of these is filamentous and closely resembles the filaments (or "microtubules") of the mitotic apparatus of other species. The larger phaneroplasmodia of two species belonging to the Physarales and the plasmodium of Hemitrichia vesparium showed fewer and less well defined fibrils, and no fibrils were seen in the aphanoplasmodium of Stemonitis fusca. Good stabilization of such fibrils in larger plasmodia may require fixation methods more rigidly controlled than those which succeed with microscopic protoplasmodia. The function of the observed fibrils cannot yet be determined. Their presence in cytoplasm fixed in situ, however, lends support to those theories of protoplasmic movement which are dependent on integral cross-bonding of one or a few molecular species.

A TEM study of the microstructure of avian eggshells

1992 ◽  
Vol 50 (2) ◽  
pp. 1102-1103
Author(s):  
S. Q. Xiao ◽  
S. Baden ◽  
A. H. Heuer
Keyword(s):  
Electron Microscope ◽  
Calcium Carbonate ◽  
Nucleation And Growth ◽  
Growth Mechanisms ◽  
Shell Surface ◽  
Thin Sections ◽  
Polycrystalline Metals ◽  
Transmission Electron ◽  
Nucleation And Growth Mechanisms

The avian eggshell is one of the most rapidly mineralizing biological systems known. In situ, 5g of calcium carbonate are crystallized in less than 20 hrs to fabricate the shell. Although there have been much work about the formation of eggshells, controversy about the nucleation and growth mechanisms of the calcite crystals, and their texture in the eggshell, still remain unclear. In this report the microstructure and microchemistry of avian eggshells have been analyzed using transmission electron microscope (TEM) and energy dispersive spectroscopy (EDS).Fresh white and dry brown eggshells were broken and fixed in Karnosky's fixative (kaltitanden) for 2 hrs, then rinsed in distilled H2O. Small speckles of the eggshells were embedded in Spurr medium and thin sections were made ultramicrotome.The crystalline part of eggshells are composed of many small plate-like calcite grains, whose plate normals are approximately parallel to the shell surface. The sizes of the grains are about 0.3×0.3×1 μm3 (Fig.l). These grains are not as closely packed as man-made polycrystalline metals and ceramics, and small gaps between adjacent grains are visible indicating the absence of conventional grain boundaries.

scholarly journals Myosin subfragment binding for the localization of actin-like microfilaments in cultured cells. A light and electron microscope study.

1977 ◽  
Vol 74 (3) ◽  
pp. 794-815 ◽  
Author(s):  
J A Schloss ◽  
A Milsted ◽  
R D Goldman
Keyword(s):  
Electron Microscope ◽  
Cultured Cells ◽  
Light And Electron Microscopy ◽  
Embryo Cell ◽  
Exchange Chromatography ◽  
Rat Embryo ◽  
Mitotic Apparatus ◽  
Thin Sections ◽  
Before And After ◽  
Identical Manner

Fluorescein-labeled heavy meromyosin subfragment-1 (F-S-1) has been purified by ion exchange chromatography and characterized in terms of its ability to bind specifically to actin. F-S-1 activates the Mg++-adenosine triphosphatase activity of rabbit skeletal muscle actin and decorates actin as shown by negative stains and thin sections of rabbit actin and rat embryo cell microfilament bundles, respectively. Binding of F-S-1 to cellular structures is prevented by pyrophosphate and by competition with excess unlabeled S-1. The F-S-1 is used in light microscope studies to determine the distribution of actin-containing structures in wnterphase and mitotic rat embryo and rat kangaroo cells. Interphase cells display the familiar pattern of fluorescent stress fibers. Chromosome-to-pole fibers are fluorescent in mitotic cells. The glycerol extraction procedures employed provide an opportunity to examine cells prepared in an identical manner by light and electron microscopy. The latter technique reveals that actin-like microfilaments are identifiable in spindles of glycerinated cells before and after addition of S-1 or HMM. In some cases, microfilaments appear to be closely associated with spindle microtubles. Comparison of the light and electron microscope results aids in the evaluation of the fluorescent myosin fragment technique and provides further evidence for possible structural and functional roles of actin in the mitotic apparatus.

scholarly journals In situ hybridization at the electron microscope level: localization of transcripts on ultrathin sections of Lowicryl K4M-embedded tissue using biotinylated probes and protein A-gold complexes.

1986 ◽  
Vol 102 (5) ◽  
pp. 1646-1653 ◽  
Author(s):  
M Binder ◽  
S Tourmente ◽  
J Roth ◽  
M Renaud ◽  
W J Gehring
Keyword(s):  
Electron Microscope ◽  
Protein A ◽  
Microscope Level ◽  
Gold Complex ◽  
Electron Microscope Level ◽  
Thin Sections ◽  
Hybrid Detection ◽  
Ultrathin Sections ◽  
Lowicryl K4m

A technique has been developed for localizing hybrids formed in situ on semi-thin and ultrathin sections of Lowicryl K4M-embedded tissue. Biotinylated dUTP (Bio-11-dUTP and/or Bio-16-dUTP) was incorporated into mitochondrial rDNA and small nuclear U1 probes by nick-translation. The probes were hybridized to sections of Drosophila ovaries and subsequently detected with an anti-biotin antibody and protein A-gold complex. On semi-thin sections, probe detection was achieved by amplification steps with anti-protein A antibody and protein A-gold with subsequent silver enhancement. At the electron microscope level, specific labeling was obtained over structures known to be the site of expression of the appropriate genes (i.e., either over mitochondria or over nuclei). The labeling pattern at the light microscope level (semi-thin sections) was consistent with that obtained at the electron microscope level. The described nonradioactive procedures for hybrid detection on Lowicryl K4M-embedded tissue sections offer several advantages: rapid signal detection: superior morphological preservation and spatial resolution; and signal-to-noise ratios equivalent to radiolabeling.

scholarly journals In situ Generation of Ruthenium Tetroxide and Osmium Tetroxide for the Physical Sciences and Their Reaction Indicators

2002 ◽  
Vol 10 (5) ◽  
pp. 20-23
Author(s):  
Paul Beauregard
Keyword(s):  
Electron Microscope ◽  
Transmission Electron Microscope ◽  
Osmium Tetroxide ◽  
Physical Sciences ◽  
Thin Sections ◽  
Ruthenium Tetroxide ◽  
Transmission Electron ◽  
In Situ Generation

Recently, there was a suggestion on the MSA listserver about the use of osmium tetroxide (OsO4 and how to handle it. One suggestion was that ampoules be scored, placed in a glass jar, and the ampoule smashed to release the contents. This seemed like a very unsafe way to use osmium tetroxide or ruthenium tetroxide. The purpose of this article is to suggest a way to generate smaller amounts of these compounds in a safer manner than smashing ampoules and wondering about what to do with the unused portion after staining or storing. Another purpose is to discuss a new reaction indicator for mainly osmium tetroxide. The use of a reaction specific indicator was mandatory for judging the level or degree to which staining had proceeded in thin sections for the transmission electron microscope (TEM).

Microanalytical evidence for short range clay coatings in weathered granodiorite

Soil Research ◽  
1981 ◽  
Vol 19 (3) ◽  
pp. 251 ◽  
Author(s):  
AJ Koppi
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Short Range ◽  
Thin Sections ◽  
Weathered Rock ◽  
Scanning Electron

Thin sections of two soils formed in situ derived from granodiorite were examined by scanning electron microscope microprobe analyser to ascertain whether clay coatings in the weathered granodiorite are illuvial or whether they are a short range phenomenon. The clay in the clay coatings of the weathered rock was of markedly different composition from the clay in the soil above. It is concluded that clay was formed from the weathering minerals in the rock and deposited as short range clay coatings within the weathered rock.

Structural Studies on Mitotic Spindle Fibres

1978 ◽  
Vol 36 (3) ◽  
pp. 615-626
Author(s):  
J.R. Mcintosh
Keyword(s):  
Chemical Composition ◽  
Mitotic Spindle ◽  
Structural Studies ◽  
Mitotic Apparatus ◽  
Chemical Studies ◽  
Medical Interest ◽  
Spindle Fibres

The mitotic apparatus is a structure of obvious biological and medical interest, but it has proved to be a difficult cellular machine to understand. The chemical composition of the spindle is only slightly elucidated, largely because of the difficulties in preparing useful isolates of the structure. Chemical studies of the mitotic spindle have been reviewed elsewhere (Mcintosh, 1977), and will not be discussed further here. One would think that structural studies on the mitotic apparatus (MA) in situ would be straightforward, but even with this approach there is some disagreement in the results obtained with various methods and by different investigators. In this paper I will review briefly the approaches which have been used in structural studies of the MA, pointing out the strengths and problems of each approach. I will summarize the principal findings of the different methods, and identify what seem to be fruitful avenues for further work.

Probing the ultrastructure of the mitotic and meiotic spindle in eggs: An expeditious approach based on semi-thin (0.25 μm) serial sections

1983 ◽  
Vol 41 ◽  
pp. 552-555
Author(s):  
Conly L. Rieder ◽  
S. Bowser ◽  
R. Nowogrodzki ◽  
K. Ross ◽  
G. Sluder
Keyword(s):  
Small Sample Size ◽  
Small Sample ◽  
Meiotic Spindle ◽  
Serial Sections ◽  
Mitotic Apparatus ◽  
Thin Sections ◽  
Cell Cycles ◽  
Ultrastructural Studies

Eggs have long been a favorite material for studying the mechanism of karyokinesis in-vivo and in-vitro. They can be obtained in great numbers and, when fertilized, divide synchronously over many cell cycles. However, they are not considered to be a practical system for ultrastructural studies on the mitotic apparatus (MA) for several reasons, the most obvious of which is that sectioning them is a formidable task: over 1000 ultra-thin sections need to be cut from a single 80-100 μm diameter egg and of these sections only a small percentage will contain the area or structure of interest. Thus it is difficult and time consuming to obtain reliable ultrastructural data concerning the MA of eggs; and when it is obtained it is necessarily based on a small sample size.We have recently developed a procedure which will facilitate many studies concerned with the ultrastructure of the MA in eggs. It is based on the availability of biological HVEM's and on the observation that 0.25 μm thick serial sections can be screened at high resolution for content (after mounting on slot grids and staining with uranyl and lead) by phase contrast light microscopy (LM; Figs 1-2).

Unusual Intranuclear Structures in Chick Sympathetic Neurons

1967 ◽  
Vol 25 ◽  
pp. 188-189
Author(s):  
E. B. Masurovsky ◽  
H. H. Benitez ◽  
M. R. Murray
Keyword(s):  
Electron Microscope ◽  
Sympathetic Neurons ◽  
Uranyl Acetate ◽  
Sympathetic Ganglia ◽  
Lead Citrate ◽  
Thin Sections ◽  
Cns Neurons ◽  
Mammalian Cns

Recent light- and electron microscope studies concerned with the effects of D2O on the development of chick sympathetic ganglia in long-term, organized culture revealed the presence of rod-like fibrillar formations, and associated granulofibrillar bodies, in the nuclei of control and deuterated neurons. Similar fibrillar formations have been reported in the nuclei of certain mammalian CNS neurons; however, related granulofibrillar bodies have not been previously described. Both kinds of intranuclear structures are observed in cultures fixed either in veronal acetate-buffered 2%OsO4 (pH 7. 4), or in 3.5% glutaraldehyde followed by post-osmication. Thin sections from such Epon-embedded cultures were stained with ethanolic uranyl acetate and basic lead citrate for viewing in the electron microscope.

Fine Structure of Interdental Cells in the Mouse Cochlea

1970 ◽  
Vol 28 ◽  
pp. 140-141
Author(s):  
Roberta M. Bruck
Keyword(s):  
Fine Structure ◽  
Electron Microscope ◽  
Young Adult ◽  
Uranyl Acetate ◽  
Ground Substance ◽  
Tectorial Membrane ◽  
Lead Citrate ◽  
Unusual Structure ◽  
Thin Sections ◽  
Collagenous Fibers

An unusual structure in the cochlea is the spiral limbus; this periosteal tissue consists of stellate fibroblasts and collagenous fibers embedded in a translucent ground substance. The collagenous fibers are arranged in vertical columns (the auditory teeth of Haschke). Between the auditory teeth are interdental furrows in which the interdental cells are situated. These epithelial cells supposedly secrete the tectorial membrane.The fine structure of interdental cells in the rat was reported by Iurato (1962). Since the mouse appears to be different, a description of the fine structure of mouse interdental cells' is presented. Young adult C57BL/6J mice were perfused intervascularly with 1% paraformaldehyde/ 1.25% glutaraldehyde in .1M phosphate buffer (pH7.2-7.4). Intact cochlea were decalcified in .1M EDTA by the method of Baird (1967), postosmicated, dehydrated, and embedded in Araldite. Thin sections stained with uranyl acetate and lead citrate were examined in a Phillips EM-200 electron microscope.

SEM Examination of a Used Diamond Knife

1971 ◽  
Vol 29 ◽  
pp. 452-453
Author(s):  
J. Temple Black
Keyword(s):  
Electron Microscope ◽  
High Voltage ◽  
High Magnification ◽  
Metallic Materials ◽  
Wide Spread ◽  
Thin Sections ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
Scanning Electron ◽  
Diamond Knife

Since its introduction by Fernandez-Moran, the diamond knife has gained wide spread usage as a common material for cutting of thin sections of biological and metallic materials into thin films for examination in the transmission electron microscope. With the development of high voltage E.M. and scanning transmission E.M., microtomy applications will become increasingly important in the preparation of specimens. For those who can afford it, the diamond knife will thus continue to be an important tool to accomplish this effort until a cheaper but equally strong and sharp tool is found to replace the diamond, glass not withstanding.In Figs. 1 thru 3, a first attempt was made to examine the edge of a used (β=45°) diamond knife by means of the scanning electron microscope. Because diamond is conductive, first examination was tried without any coating of the diamond. However, the contamination at the edge caused severe charging during imaging. Next, a thin layer of carbon was deposited but charging was still extensive at high magnification - high voltage settings. Finally, the knife was given a light coating of gold-palladium which eliminated the charging and allowed high magnification micrographs to be made with reasonable resolution.

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