Ultrastructural differences in the exosporium of the Sterne and Vollum strains of Bacillus anthracis

1968 ◽  
Vol 14 (12) ◽  
pp. 1297-1299 ◽  
Author(s):  
M. J. Kramer ◽  
Ivan L. Roth
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Bacillus Anthracis ◽  
Potassium Permanganate ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Single Application ◽  
Thin Sections ◽  
Acetate Lead ◽  
Different Strains

Spores from three different strains of Bacillus anthracis were examined by electron microscopy for the presence of a hair-like nap previously reported to be present on the exosporium of spores of the Sterne strain (avirulent). In addition to that strain, the Vollum strain (virulent) and a rough, avimlent variant of the Vollum were utilized in the current studies.Spores were fixed with Kellenberger's standard OsO4 fixative and embedded in Maraglas. Thin sections were poststained with various combinations of the following: potassium permanganate, uranyl acetate, lead citrate. The nap on the exosporium of spores of the Sterne strain was revealed most clearly when thin sections were poststained with all of the aforementioned stains. Post-staining by a single application of any of the three reagents resulted in a nap that was barely perceptible.The surface of the exosporium of spores from the Vollum strain and the rough, avirulent variant was found to be quite different from that of the Sterne strain. On the two former, the surface layer is approximately one-third as thick as the layer of hairs in the nap on the latter.

A double staining technique for improved contrast of thin sections from Spurr-embedded tissue

1978 ◽  
Vol 56 (1) ◽  
pp. 129-132 ◽  
Author(s):  
D. F. Bray ◽  
E. B. Wagenaar
Keyword(s):  
Potassium Permanganate ◽  
Staining Technique ◽  
Uranyl Acetate ◽  
Staining Procedure ◽  
Lead Citrate ◽  
Double Staining ◽  
Thin Sections ◽  
Citrate Method ◽  
Acetate Lead

A double staining procedure using potassium permanganate and lead citrate is described. The technique, involving treatment of sections with a 1% solution of potassium permanganate followed by lead citrate staining, is shown to be more effective in producing contrast in Spurr-embedded tissue than the conventional uranyl acetate – lead citrate method.

Ultrahistochemical Analysis of the Chick Embryo Cornea Using A Non-Dispersive X-Ray Detector

1972 ◽  
Vol 30 ◽  
pp. 404-405
Author(s):  
T. Ohkura ◽  
M. Takashio ◽  
T. Watanabe
Keyword(s):  
Electron Microscopy ◽  
Chick Embryo ◽  
Alcian Blue ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Thin Sections ◽  
X Ray ◽  
Transmission Electron ◽  
Glutaraldehyde Solution ◽  
Tissue Specimens

The experiments which we report here were performed to show qualitatively the presence or abscence of Cu in the mesostroma of the chick embryo cornea stained with alcian blue 8GS. Strips of the cornea were fixed in a buffered glutaraldehyde solution (2.5%, pH 7.4) and coloured with alcian blue 8GS at pH 2.5. Tissue specimens were postosmicated, dehydrated and embedded in Epon. Tissue blocks for the conventional transmission electron microscopy were prepared without alcian blue treatment, and the thin sections were stained with uranyl acetate and/or lead citrate. Fig. la shows the mesostroma of 3rd day chick embryo cornea; filaments run in various directions, and reveal no visible periodicity. Interfibrillar substances can not be demonstrated by the conventional method of the electron staining. The alcian blue treatment reveals the interfibrillar substances, which stand out in contrast as shown in Fig. lb.

The Surface Structure of Cultured Rabbit Kidney Cells as Revealed by Electron Microscopy

1970 ◽  
Vol 7 (3) ◽  
pp. 719-737
Author(s):  
ELIZABETH DIMMOCK
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Cell Surface ◽  
Blood Group ◽  
Potassium Permanganate ◽  
Ruthenium Red ◽  
Rabbit Kidney ◽  
Dense Layer ◽  
Kidney Cells ◽  
Thin Sections

An epithelioid line of rabbit kidney cells (RK 13), in which the distribution of blood group antigen A had previously been investigated by mixed agglutination, was chosen for studies of the structure of the cell surface. Cells were grown in monolayer culture, and thin sections were examined by electron microscopy after fixation in glutaraldehyde and osmium tetroxide and staining of the sections with lead, or uranyl and lead. Cells were also treated with ruthenium red incorporated in the osmium fixative. Other cells were fixed in lanthanum or potassium permanganate, and both stained and unstained sections were examined. The morphology of RK 13 cell surfaces is described. There is apparently no great degree of cellular specialization. The treatment with ruthenium red resulted in dense staining of a layer of the cell surface that is not visible in conventional preparations, and sometimes in staining of the surfaces of intracellular organelles. Lanthanum permanganate fixation also revealed a dense layer on the surface of the plasma membrane; a less dense surface layer was distinguished in many cells fixed in potassium permanganate. The reaction of ruthenium red with the cell surface is probably due to the presence of acidic glycoproteins, but the chemical specificity of the staining method is not yet clear. The nature of the material revealed by lanthanum and potassium permanaganates is also undefined. However, these staining methods reveal that the cell surface is more complex than is apparent in cells prepared by conventional techniques. The additional surface layer is probably the site of many blood group substances and other compounds involved in the physiological reactions of the cell surface.

Electron microscopic evidence for a double hair-like nap appearing at low frequency on Bacillus anthracis Sterne spores

1969 ◽  
Vol 15 (10) ◽  
pp. 1247-1248 ◽  
Author(s):  
M. J. Kramer ◽  
Ivan L. Roth
Keyword(s):  
Bacillus Anthracis ◽  
Microscopic Examination ◽  
Low Frequency ◽  
Electron Microscopic Examination ◽  
Uranyl Acetate ◽  
Spore Coat ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Very Low Frequency

Electron microscopic examination of thin sections of Bacillus anthracis Sterne spores triply poststained with KMnO4, uranyl acetate, and lead citrate has indicated an unusual morphological variant. These spores are seen at very low frequency and have, in addition to the hair-like nap normally associated with the exosporium a second hairy layer which appears to originate in the spore coat complex.

EM Study of Isopod Hemocytes

1998 ◽  
Vol 4 (S2) ◽  
pp. 1138-1139
Author(s):  
G. M. Vernon ◽  
E. J. Rappa ◽  
W. C. Murray ◽  
R. Witkus
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Morphological Analysis ◽  
Standard Technique ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Small Granule ◽  
Thin Sections ◽  
Cytoplasmic Granules ◽  
Transmission Electron

Crustacean hemocytes have been characterized on the basis of cell size and nature of cytoplasmic granules. Based on light microscopic morphological analysis and cytochemistry, investigators variously named the hemocyte types (agranular, small-granule, large granule, undifferentiated, hyaline cells, non-explosive, explosive granulocytes, etc.). In his study of the isopod Armadillidium vulgare Faso adopted the terminology of Benjamin and James and referred to the hemocytes as hyaline cells, semi-granulocytes and granulocytes.In the present investigation we have studied the hemocytes of two isopods, Oniscus asellus and Armadillidium nasatum, using transmission electron microscopy. Hemolymph was collected by penetrating the posterior dorsal exoskeleton of 20 animals of each genus with a microcapillary pipette and drawing 3-5μL per isopod. The samples were processed following a standard technique. Thin sections were collected on 300 mesh copper grids, counterstained with 2% aqueous uranyl acetate and lead citrate, and viewed with a JEOL 1010 electron microscope.

Morphometric comparison of uranyl acetate/lead citrate-stained and peroxidase-stained neutrophil granules

1990 ◽  
Vol 48 (3) ◽  
pp. 756-757
Author(s):  
Charles S. Gilbert ◽  
Richard T. Parmley ◽  
Joseph M. Kinkade
Keyword(s):  
Human Peripheral Blood ◽  
Uranyl Acetate ◽  
Buffy Coat ◽  
Significant Heterogeneity ◽  
Lead Citrate ◽  
Thin Sections ◽  
Low Viscosity ◽  
Physical Density ◽  
Acetate Lead ◽  
Blood Neutrophils

Although neutrophil granules can be broadly divided into peroxidase-positive and peroxidasenegative granules, recent studies have demonstrated significant heterogeneity in neutrophil granule morphology and physical density. The present study was undertaken to evaluate this heterogeneity morphometrically. Human peripheral blood neutrophils were collected from normal volunteers in heparin or EDTA and fixed in 3% glutaraldehyde as a cell suspension or minced buffy coat. An aliquot was stained for myeloperoxidase using 3,3’-diaminobenzidine (DAB) as substrate. All samples were post fixed in 1% OSO4 and embedded in Spurr low viscosity medium. Additionally neutrophil granules isolated according to the method of Rice, et al. were similarly embedded. Thin sections of morphologic preparations were counterstained with methanolic uranyl acetate and aqueous lead citrate (UALC) (Fig. 1), whereas DAB stained specimens were counterstained with LC only (Fig. 2). Attempts to combine UALC and DAB staining in the same section obscured the density imparted by DAB. LC failed to impart significant density to DAB-negative granules in DAB-LC stained specimens resulting in an underestimation of this granule population.

On the Location of Stain in Mitochondria

1968 ◽  
Vol 26 ◽  
pp. 128-129
Author(s):  
G. B. Haydon ◽  
S. O. Smith
Keyword(s):  
Skeletal Muscle ◽  
Heavy Metal ◽  
Oxidative Phosphorylation ◽  
Focal Plane ◽  
Uranyl Acetate ◽  
Thick Section ◽  
Lead Citrate ◽  
Thin Sections ◽  
Thin Sectioning ◽  
Acetate Lead

The degree of matrical staining in isolated and tissue mitochondria has been shown to be a function of their state of oxidative-phosphorylation at the time of fixation (J. Histochem. Cytochem. 15:752, 1967). High resolution EM in conjunction with these studies revealed amplitude contrast images 10 AU and larger in diameter which are independent of the focal plane. These amplitude densities have been interpreted as individual aggregates of heavy metal stain (J. Cell Biol. 35:55A, 1967). Two approaches have been used in an effort to locate such stain aggregates.(1) Four micron epoxy sections of skeletal muscle, cardiac muscle, and liver were stained with uranyl acetate, lead citrate, or both, following the methods used to stain 500 AU sections for EM. After staining, the thick epon sections were re-embedded and thin sections cut perpendicular to the stained surface. Either or both of these metal stains penetrate 500 to 1000 AU into the thick section and give the tissue increased contrast similar to that due to staining after thin sectioning. The mitochondrial matrices are lightly stained indicating that they are not in a state of active oxidative-phosphorylation. Figure 1 shows such mitochondria at low magnification before (Figure 1a) and after (Figure lb) restaining with uranyl acetate and lead citrate. After restaining it is no longer possible to identify the original stained surface.

scholarly journals Electron Microscopy of In-Plaque Phage T3 Assembly: Proposed Analogs of Neurodegenerative Disease Triggers

2020 ◽  
Vol 13 (1) ◽  
pp. 18 ◽  
Author(s):  
Philip Serwer ◽  
Barbara Hunter ◽  
Elena T. Wright
Keyword(s):  
Electron Microscopy ◽  
Innate Immunity ◽  
Neurodegenerative Disease ◽  
Virus Assembly ◽  
Uranyl Acetate ◽  
Host Responses ◽  
Thin Sections ◽  
Viral Capsids ◽  
Acetate Lead ◽  
Forming Temperature

Increased knowledge of virus assembly-generated particles is needed for understanding both virus assembly and host responses to virus infection. Here, we use a phage T3 model and perform electron microscopy (EM) of thin sections (EM-TS) of gel-supported T3 plaques formed at 30 °C. After uranyl acetate/lead staining, we observe intracellular black particles, some with a difficult-to-see capsid. Some black particles (called LBPs) are larger than phage particles. The LBP frequency is increased by including proflavine, a DNA packaging inhibitor, in the growth medium and increasing plaque-forming temperature to 37 °C. Acidic phosphotungstate-precipitate (A-PTA) staining causes LBP substitution by black rings (BRs) that have the size and shape expected of hyper-expanded capsid containers for LBP DNA. BRs are less frequent in liquid cultures, suggesting that hyper-expanded capsids evolved primarily for in-gel (e.g., in-biofilm) propagation. BR-specific A-PTA staining and other observations are explained by α-sheet intense structure of the major subunit of hyper-expanded capsids. We hypothesize that herpes virus triggering of neurodegenerative disease occurs via in-gel propagation-promoted (1) generation of α-sheet intense viral capsids and, in response, (2) host production of α-sheet intense, capsid-interactive, innate immunity amyloid protein that becomes toxic. We propose developing viruses that are therapeutic via detoxifying interaction with this innate immunity protein.

Lead aspartate: a new en bloc stain for electron microscopy

1978 ◽  
Vol 36 (2) ◽  
pp. 34-35
Author(s):  
J.R. Walton
Keyword(s):  
Electron Microscopy ◽  
Calcium Ion ◽  
Enzyme Reaction ◽  
Lead Citrate ◽  
Reaction Products ◽  
En Bloc ◽  
Thin Sections ◽  
Lead Salts ◽  
Thin Sectioning ◽  
High Alkalinity

In electron microscopy, lead is the metal most widely used for enhancing specimen contrast. Lead citrate requires a pH of 12 to stain thin sections of epoxy-embedded material rapidly and intensively. However, this high alkalinity tends to leach out enzyme reaction products, making lead citrate unsuitable for many cytochemical studies. Substitution of the chelator aspartate for citrate allows staining to be carried out at pH 6 or 7 without apparent effect on cytochemical products. Moreover, due to the low, controlled level of free lead ions, contamination-free staining can be carried out en bloc, prior to dehydration and embedding. En bloc use of lead aspartate permits the grid-staining step to be bypassed, allowing samples to be examined immediately after thin-sectioning.Procedures. To prevent precipitation of lead salts, double- or glass-distilled H20 used in the stain and rinses should be boiled to drive off carbon dioxide and glassware should be carefully rinsed to remove any persisting traces of calcium ion.

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