Ultrastructural aspects of the cell envelope of Legionella

1984 ◽  
Vol 42 ◽  
pp. 220-221
Author(s):  
C.N. Sun ◽  
P. Morgan ◽  
W.R. Bowen
Keyword(s):  
Osmium Tetroxide ◽  
Cell Envelope ◽  
Bacterial Species ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Ultrastructural Organization ◽  
Thin Sections ◽  
Gram Negative ◽  
Additional Information ◽  
Legionnaires Disease

The causal agents of Legionnaires’ Disease and related respiratory ailments in humans are gram-negative rod-shaped bacterial species belonging to the newly recognized genus, Legionella. During a comparative study of the ultrastructural organization of the cell boundary in several species, additional information regarding the presence and nature of various appendages of the cell envelope in these bacteria was observed.Cultures of L. pneumophils (including several serogroups) and L. micdadei were grown on charcoal yeast extract agar with increased humidity at 37°C. Samples were removed after 5 days of growth and immediately fixed in either glutaraldehyde/ruthenium red (GA/RR) followed by osmium tetroxide/ruthenium red (OSO4/RR) or glutaraldehyde/tannic acid (GA/TA) followed by osmium tetroxide. Thin sections were stained with uranyl acetate and lead citrate.

Ultrastructural Characteristics of Vaginal Epithelium of Persistent Estrous Rats

1985 ◽  
Vol 43 ◽  
pp. 658-659
Author(s):  
Khosho Francis K. ◽  
Kaufmann Robert C. ◽  
Amankwah Kofi S.
Keyword(s):  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Female Rats ◽  
Constant Light ◽  
Ultrastructural Changes ◽  
Vaginal Smears ◽  
Thin Sections ◽  
Cacodylate Buffer ◽  
Ultrastructural Characteristics

Adult female rats exposed to constant light will develop anovulatory acyclicity characterized by persistent vaginal cornification (PE) and formation of multiple large cystic follicles on the ovaries. The purpose of the present communication is to describe the ultrastructural changes in vaginal epithelia in PE rats as compared to that in normal estrous rats.Persistent vaginal estrous with PCO was induced in a group of Sprague-Dawely rats by exposure to constant light for 50-150 days. Rats in normal estrous, as determined by vaginal smears, were used as controls. Nembutal- anethesized rats were perfused through the aorta with 2.5% gluteraldehyde in 1M sodium cacodylate buffer (pH 7.3). The mucosa of the vaginal folds just inferior to the cervix were dissected by microsurgery, postfixed, stained with 0.5% ruthenium red in 1% osmium tetroxide, dehydrated, and embedded in polybed. Thick sections (1μ) were stained with toludine blue for light microscopy studies. Thin sections were stained with uranyl acetate and lead citrate.

The response of cell walls of Bacillus subtilis to metals and to electron-microscopic stains

1978 ◽  
Vol 24 (2) ◽  
pp. 89-104 ◽  
Author(s):  
T. J. Beveridge
Keyword(s):  
Bacillus Subtilis ◽  
Electron Scattering ◽  
Cell Walls ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Transition Elements ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Wall Ultrastructure

Purified cell walls of Bacillus subtilis were subjected to solutions of 40 independent metals and the metal uptake, the electron-scattering power of thin sections, and the type of staining response evaluated. This was repeated for six typical electron-microscopic stains (uranyl acetate, uranyl magnesium acetate, osmium tetroxide, Os-meth, osmium-dimethylethylenediamine, and ruthenium red) and one new staining reagent (a potassium platinum chloride – dimethylsulfoxide complex) whose specificity is for amine functions. The reaction of select metals can be specific in terms of both uptake and staining response. Of the metals studied most transition elements had a high affinity for the wall fabric and some (i.e., Sc III, most lanthanides, U IV, Zr IV, Hf IV, Fe III, Pd II, Ru III, and In III) may be suitable as contrasting agents for electron microscopy. Furthermore, when the thickness of metal-reacted walls was compared to freeze-each and ultracryotomy data, statistical-dimensional differences were commonly seen, which indicates that wall ultrastructure can be profoundly affected by the type of metal and (or) staining reagent.

Membranous alterations in the cytoplasm and nucleus in a primitive neuroectodermal tumor of the brain

1978 ◽  
Vol 36 (2) ◽  
pp. 628-629
Author(s):  
C. N. Sun ◽  
C. Araoz ◽  
H. J. White
Keyword(s):  
Nuclear Envelope ◽  
Tumor Cells ◽  
Osmium Tetroxide ◽  
Primitive Neuroectodermal Tumor ◽  
Uranyl Acetate ◽  
Neuroectodermal Tumor ◽  
Annulate Lamellae ◽  
Lead Citrate ◽  
Thin Sections ◽  
The Brain

The ultrastructure of a cerebral primitive neuroectodermal tumor has been reported previously. In the present case, we will present some unusual previously unreported membranous structures and alterations in the cytoplasm and nucleus of the tumor cells.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4% glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1% buffered osmium tetroxide for one hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate.In the cytoplasm of the tumor cells, we found paired cisternae (Fig. 1) and annulate lamellae (Fig. 2) noting that the annulate lamellae were sometimes associated with the outer nuclear envelope (Fig. 3). These membranous structures have been reported in other tumor cells. In our case, mitochondrial to nuclear envelope fusions were often noted (Fig. 4). Although this phenomenon was reported in an oncocytoma, their frequency in the present study is quite striking.

Ultrastructure of myoepithelial cell in parotid gland

1988 ◽  
Vol 46 ◽  
pp. 342-343
Author(s):  
C. N. Sun
Keyword(s):  
Parotid Gland ◽  
Osmium Tetroxide ◽  
Individual Cell ◽  
Branching Processes ◽  
Muscle Cells ◽  
Myoepithelial Cells ◽  
Uranyl Acetate ◽  
Thin Sections ◽  
Gland Carcinoma ◽  
Parotid Gland Carcinoma

Myoepithelial cells have been observed in the prostate, harderian, apocrine, exocrine sweat and mammary glands. Such cells and their numerous branching processes form basket-like structures around the glandular acini. Their shapes are quite different from structures seen either in spindleshaped smooth muscle cells or skeletal muscle cells. These myoepithelial cells lie on the epithelial side of the basement membrane in the glands. This presentation describes the ultrastructure of such myoepithelial cells which have been found also in the parotid gland carcinoma from a 45-year old patient.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4 percent glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1 percent buffered osmium tetroxide for 1 hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate. Ultrastructurally, the pattern of each individual cell showed wide variations.

Observations of thick and thin sections in a field-emission SEM

1994 ◽  
Vol 52 ◽  
pp. 1018-1019
Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley
Keyword(s):  
Electron Microscope ◽  
Field Emission ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Transmission Electron ◽  
Conventional Transmission Electron Microscope ◽  
Scanning Electron

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.

ELECTRONMICROSCOPIC STUDIES ON PLATELETS AND MEGAKARYOCYTES IN GIANT PLATELET SYNDROME

1987 ◽  
Author(s):  
Sozo Suzuki ◽  
Kazuo Mori ◽  
Koji Sugai ◽  
Yasuyuki Akutsu ◽  
Masaaki Ishikawa ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Membrane Systems ◽  
Platelet Factor ◽  
Giant Platelets ◽  
Thin Sections ◽  
Giant Platelet ◽  
Dense Bodies ◽  
Large Platelet

Giant platelet syndrome are characterized morphologically by many giant platelets associated with several functional abnormalities in the peripheral blood. However, the mechanism of large platelet production has not yet been clarified. In 1981, we reported acase with Bernard-Soulier syndrome(BSS) in whom giant platelets were considered to be formed by fusion of two or three platelets in the circulating blood. We examined the ultrastructure of platelets and megakaryocytes in another case with BSS (29 year-old female) and a case with May-Hegglin anomaly (31 year-old male). Whole blood and bone marrow specimens were fixed with glutaraldehyde-osmium solution. Thin sections were prepared and stained with uranyl acetate and lead cytrate. Membrane systems of platelets and megakaryocytes in a case with BSS was investigated by staining of surface coating with ruthenium red.In a case with BSS, most platelets were very large and similar in morphology to those in formerly reported case. Giant platelets contained several-fold increased number of α-granules and mitochondria. Typical dense bodies were also observed. Contents of ATP/ADP, platelet factor-4(PF-4), B-thromboglobulin(B-TG) and platelet factor-3 availability(PF-3) were increased. Disorganization of microtubules was recognized. Some giant platelet contained membrane systems similar to demarcation membranes(DM) in megakaryocytes, characteristically. In mature megakaryocytes, areas divided by DM similar in size to those in normal megakaryocytes were observed. Several of these areas appeared to fuse together to form the giant platelets containing many granules and remnants of DM. In a case with May-Hegglin anomaly, typical Dohle’s bodies were shown in neutrophilic granulocytes. Giant platelets in this case also contained large number of α-granules and some of them contained membrane systems similar to DM. Areas similar in morphology to these giant platelets were clearly noted in the cytoplasm of mature megakaryocytes.In these cases, most giant platelets in the peripheral blood may be formed in the cytoplasm of megakaryocytes by fusion of several areas divided by DM, each of which may become normal sized platelets in normal megakaryocytes.

scholarly journals Immunocytochemical Localization of a Calmodulinlike Protein in Bacillus subtilis Cells

1999 ◽  
Vol 181 (15) ◽  
pp. 4605-4610 ◽  
Author(s):  
Delfina C. Dominguez ◽  
Hank Adams ◽  
James H. Hageman
Keyword(s):  
Bacillus Subtilis ◽  
Cellular Localization ◽  
Time Course ◽  
Cell Envelope ◽  
Membrane Vesicles ◽  
Cross Reactivity ◽  
Bovine Brain ◽  
Ruthenium Red ◽  
Thin Sections ◽  
Calcium Ion Transport

ABSTRACT To determine possible functions of the calmodulinlike protein ofBacillus subtilis, the time course of its expression during sporulation and its cellular localization were studied. The protein was expressed in a constitutive manner from the end of logarithmic growth through 8 h of sporulation as determined by antibody cross-reactivity immunoblots and enzyme-linked immunosorbent assays (ELISAs). In partially purified extracts, the immunopositive protein comigrated upon electrophoresis with a protein which selectively bound [45Ca]CaCl2, ruthenium red, and Stains-all. Previous studies showed increased extractability of the calmodulinlike protein from B. subtilis cells when urea and 2-mercaptoethanol were used in breakage buffers, implying that the protein might be partially associated with the membrane fraction. This was confirmed by demonstrating that isolated membrane vesicles ofB. subtilis also gave positive immunological tests with Western blotting and ELISAs. To more precisely locate the protein in cells, thin sections of late-log-phase cells, sporulating cells, and free spores were reacted first with bovine brain anticalmodulin specific antibodies and then with gold-conjugated secondary antibodies; the thin sections were examined by transmission electron microscopy. The calmodulinlike protein was found almost exclusively associated with the cell envelope of these fixed, sectioned cells. A possible function of the calmodulinlike protein in sensing calcium ions or regulating calcium ion transport is suggested.

An Electron Microscopic Investigation of the Pathogenesis of Hemorrhage Induced by Rattlesnake Venom

1974 ◽  
Vol 32 ◽  
pp. 56-57
Author(s):  
Charlotte L. Ownby ◽  
Robert A. Kainer ◽  
Anthony T. Tu
Keyword(s):  
Phosphate Buffer ◽  
Osmium Tetroxide ◽  
Microscopic Investigation ◽  
Uranyl Acetate ◽  
Electron Microscopic Investigation ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Rattlesnake Venom ◽  
Diamondback Rattlesnake

One of the significant changes induced by the injection of rattlesnake (Crotalidae) venom is hemorrhage. Since crotaline antivenin does not prevent such local tissue damage, a more effective treatment of snakebite is needed. To aid in the development of such a treatment the pathogenesis of venom-induced hemorrhae was investigated.Swiss-Webster white mice were injected intramuscularly with Western diamondback rattlesnake (Crotalus atrox) venom. Two minutes after the injection, muscle tissue was obtained by bioosy from the thigh and fixed in 6% glutaraldehyde in Milloniq's phosphate buffer (DH 7.4, 2 hrs., 4°C). After post-fixation in 2% osmium tetroxide in Milloniq's phosphate buffer (pH 7.4, 1hr., 4°C) the tissue was dehydrated routinely in ethanol and embedded in Epon 812. The thin sections were stained with uranyl acetate in methanol and lead citrate then observed with either a Zeiss EM 9A or an Hitachi HS-8 electron microscope.

Ultrastructure of Granules Passing Through Frog Oocyte Nuclear Pores

1969 ◽  
Vol 27 ◽  
pp. 288-289
Author(s):  
Madison B. Cole
Keyword(s):  
Osmium Tetroxide ◽  
Rana Pipiens ◽  
Uranyl Acetate ◽  
Nuclear Pores ◽  
Thin Sections ◽  
Close Proximity ◽  
Perinuclear Cytoplasm ◽  
Frog Oocyte ◽  
Pass Through ◽  
Number Of Authors

Ovaries of Rana pipiens larvae were fixed in glutaraldehyde, post-fixed in osmium tetroxide, and embedded in epon. Thin sections of oocytes were routinely stained using methanolic uranyl acetate followed by lead citrate.The granule, designated M1, found in the primary oocytes is considered to pass through the nuclear pores by virtue of 1) its location in the nucleoplasm in close proximity to the nuclear envelope, 2) its contact with the nuclear pores, 3) its presence in aggregates in the perinuclear cytoplasm and A) its similarity to granules referred to by a number of authors (1-4) as material passing through nuclear pores.

Comparison of Three Common Fixatives on Bacteria

1977 ◽  
Vol 35 ◽  
pp. 348-349
Author(s):  
J.M. Hanson ◽  
R.M. Pfister ◽  
R.A. Smucker
Keyword(s):  
Bacillus Megaterium ◽  
Cell Walls ◽  
Uranyl Acetate ◽  
Ruthenium Red ◽  
Thin Sections ◽  
Internal Morphology ◽  
Cytoplasmic Membranes ◽  
Glutaraldehyde Solution ◽  
Background Material ◽  
Whole Mounts

The ultrastructure of the cytoplasmic membranes, cell walls, and mesosomes of Escherichia coli and Bacillus megaterium were found to be significantly affected by different fixation procedures.Logarithmic cells were divided into equal portions and different fixation procedures employed. A 6% glutaraldehyde solution followed by 1% 0s04 fixa-tion (1). The Ryter-Kellenberger (R-K) fixation (2) and Luft's ruthenium red (R-R) (3). The glutaraldehyde-OsO4 and R-K were post fixed with 2% and 0.5% uranyl acetate respectively. Thin sections were post stained with uranylacetate and lead citrate. One sub-portion of Bacillus megaterium was criti-cally-point dried. Glutaraldehyde-OsO4 critically-point dried whole mounts (Fig. 1) showed little background material and smooth walls. The R-K cells (Fig. 2) had irregular outer walls. The R-R cells (Fig. 3) were irregular and revealed external polysaccharide. There is a distinct difference in the internal morphology of the wall layers using the E. coli (Figs. 4,5,6). A heavier staining of the CM was observed in B. megaterium (Fig. 7).

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