scholarly journals Ultrastructure of Lipid Deposits and Other Contents in Freeze-Etched Coleoptile Cells of Ungerminated Rice Grains

1973 ◽  
Vol 26 (2) ◽  
pp. 357 ◽  
Author(s):  
MS Buttrose ◽  
A Soeffky
Keyword(s):  
Aqueous Media ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Rice Grains ◽  
Freeze Etching

Freeze-etching has been used to avoid exposure to aqueous media in studying ultrastructure in coleoptile cells of ungerminated rice grains. Results differ in some respects from those obtained after chemical fixation and examination of thin sections.

Fine structure of the macroconidia of Fusarium solani

1975 ◽  
Vol 53 (19) ◽  
pp. 2134-2146 ◽  
Author(s):  
J. P. Tewari ◽  
W. P. Skoropad
Keyword(s):  
Cell Wall ◽  
Fusarium Solani ◽  
Superficial Layer ◽  
Dense Layer ◽  
Inoculum Potential ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Innermost Layer ◽  
Freeze Etching ◽  
Myelin Figures

Ultrastructure of the macroconidia of Fusarium solani as visualized by transmission (ultrathin sectioning and freeze-etching) and scanning electron microscopy is described. The cell wall has four layers. The innermost layer is electron-lucid followed by an electron-dense layer. The next outer layer is spongy in appearance followed by a superficial layer consisting of fine filamentous processes. Freeze-etch replicas of conidia directly removed from the sporodochia and still suspended in the mucilaginous material (in which they are produced) frequently show the conidia connected by the superficial filamentous processes in the cell wall. This agglutination of the conidia is likely to increase the inoculum potential of this pathogen at the sites of infection. Structure of various membrane systems in the cells is described. The endoplasmic reticulum is fairly extensive and fenestrated. Thin sections of routinely fixed conidia show myelin figures. However, such structures were not seen in replicas of conidia that were freeze-etched without use of chemical fixation or cryoprotection.

Mesosome-like Structures in Cryptococcus neoformans Studied by Chemical Fixation and by the Freeze-etching Technique

1968 ◽  
Vol 26 ◽  
pp. 80-81
Author(s):  
M. R. Edwards ◽  
S. C. Holt
Keyword(s):  
Electron Microscope ◽  
Cryptococcus Neoformans ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Chemical Fixation ◽  
Etching Technique ◽  
Pathogenic Yeast ◽  
Thin Sections ◽  
Lead Salts ◽  
Freeze Etching

The general features of Cryptococcus neoformans, a pathogenic yeast, have been studied with the electron microscope. In the course of such a study it was noted that the plasma membrane of C. neoformans, occasionally invaginated into the cytoplasm and formed membranous organelles which resembled bacterial mesosomes. The present investigation was undertaken in order to examine such structures in detail and to compare the results from chemical fixation with those of freeze-etching.Cells were grown in Sabouraud's agar at 25-27° C for 24-48 hr and fixed with 4% glutaraldehyde in 0.15 M phosphate (Sbrensen's) buffer, at room temperature, for 2 hr; after being thoroughly washed in the buffer and post-fixed in osmium tetroxide, in the same buffer, they were dehydrated in ethyl alcohol and embedded in Epon. Thin sections were cut in a LKB microtome, double stained with uranyl and lead salts and examined in the Siemens Elmiskop IA.

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.

Freeze substitution fixation of fungal reproductive structures and spores

1989 ◽  
Vol 47 ◽  
pp. 990-991
Author(s):  
C. W. Mims ◽  
E. A. Richardson
Keyword(s):  
Plant Pathogens ◽  
Freeze Substitution ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Dialysis Membrane ◽  
Razor Blade ◽  
Thin Sections ◽  
Reproductive Structures ◽  
Dry Down ◽  
Diamond Knife

The advantages of freeze substitution fixation over conventional chemical fixation for preservation of ultrastructural details in fungi have been discussed by various authors. As most ascomycetes, basidiomycetes and deuteromycetes do not fix well using conventional chemical fixation protocols, freeze substitution has attracted the attention of many individuals interested in fungal ultrastructure. Thus far most workers using this technique on fungi have concentrated on thin walled somatic hyphae. However, in our laboratory we have experimented with the use of freeze substitution on a variety of fungal reproductive structures and spores with promising results.Here we present data on freeze substituted samples of sporangia of the zygomycete Umbellopsis vinacea, basidia of Exobasidium camelliae var. gracilis, developing teliospores of the smut Sporisorium sorghi, germinating teliospores of the rust Gymnosporangium clavipes, germinating conidia of the deuteromycete Cercosporidium personatum, and developing ascospores of Ascodesmis nigricans.Spores of G. clavipes and C. personatum were deposited on moist pieces of sterile dialysis membrane where they hydrated and germinated. Asci of A. nigricans developed on pieces of dialysis membrane lying on nutrient agar plates. U. vinacea was cultured on small pieces of agar-coated wire. In the plant pathogens E. camelliae var. gracilis and S. sorghi, a razor blade was used to remove smal1 pieces of infected host issue. All samples were plunged directly into liquid propane and processed for study according to Hoch.l Samples on dialysis membrane were flat embedded. Serial thin sections were cut using a diamond knife, collected on slot grids, and allowed to dry down onto Formvar coated aluminum racks. Sections were post stained with uranyl acetate and lead citrate.

Ultrastructural comparisons of fungus hyphal cells using frozen-etched replicas and thin sections of the fungus Pyrenochaeta terrestris

1968 ◽  
Vol 14 (3) ◽  
pp. 205-210 ◽  
Author(s):  
W. M. Hess
Keyword(s):  
Electron Scattering ◽  
Cell Walls ◽  
Sole Carbon Source ◽  
Membrane Systems ◽  
Thin Sections ◽  
Fixed Membrane ◽  
Pyrenochaeta Terrestris ◽  
Freeze Etching ◽  
Fixed Cell ◽  
Etched Membrane

The ultrastructure of P. terrestris hyphal cells was investigated to compare frozen-etched replicas with chemically fixed thin sections. The fungus used in this study uses glycerol as a sole carbon source and survives the freezing procedures necessary for freeze-etching; thus frozen-etched replicas reflect the living state.Frozen-etched membrane systems have particles of various sizes and concentrations and have a smooth appearance as contrasted to chemically fixed membrane systems, which have particles difficult to distinguish and somewhat irregular membrane systems. Frozen-etched cell walls are seen to contain particles, and microfibrillar orientation is evident in older cell walls, whereas substructure is not evident in chemically fixed cell walls, although secretion products of the fungus accumulate on cell surfaces.Chemically fixed ground cytoplasm has ribosomes and areas of high- and low-electron scattering which are not seen with freeze-etching. Cells fixed in glutaraldehyde–acrolein–OsO4 more nearly resemble frozen-etched cells than cells fixed in potassium permanganate.

Fine structure of the thermophilic blue-green alga Synechococcus lividus Copeland. A study of frozen–fractured–etched cells

1972 ◽  
Vol 18 (2) ◽  
pp. 175-181 ◽  
Author(s):  
S. C. Holt ◽  
M. R. Edwards
Keyword(s):  
Green Alga ◽  
Lipid Bodies ◽  
Blue Green Alga ◽  
Etching Technique ◽  
Thin Sections ◽  
Blue Green Algae ◽  
Osmiophilic Granules ◽  
Freeze Etching ◽  
The Many ◽  
Synechococcus Lividus

The thermophilic unicellular blue-green alga, Synechococcus lividus, was studied by electron microscopy in thin sections and by the freeze-etching technique. Thin sections revealed subcellular structures like those observed by other authors in mesophilic blue-green algae. In the freeze-etched fractures similar results were obtained but, in addition, surface views of plasma and thylakoidal membranes were examined in detail. The many inclusions present in the freeze-etched preparations confirmed those displayed in thin sections and are interpreted as polyhedral, polyphosphate, and lipid bodies. Some unidentified osmiophilic granules and also phycobilisomes were seen.

scholarly journals ULTRASTRUCTURE AND CALCIUM TRANSPORT IN CRUSTACEAN MUSCLE MICROSOMES

1971 ◽  
Vol 48 (1) ◽  
pp. 49-60 ◽  
Author(s):  
R. J. Baskin
Keyword(s):  
Freeze Drying ◽  
Calcium Uptake ◽  
Membrane Thickness ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Crustacean Muscle ◽  
Thin Sectioning ◽  
Critical Point Drying ◽  
Freeze Etching ◽  
Microscopic Techniques

Fragmented sarcoplasmic reticulum (FSR) from crustacean muscle was examined following preparation by a variety of electron microscopic techniques. The 30–40 A particles which appeared on the outer surface of FSR vesicles following negative staining were not observed following preparation by freeze-drying, freeze-etching, thin sectioning, or critical-point drying. Crustacean FSR exhibited high values of calcium uptake and extensive nodular formation in the presence of oxalate. 80–90 A diameter membrane particles were seen in freeze-etch preparations of both intact lobster muscle and FSR vesicles. Thin sections of FSR vesicles revealed a membrane thickness of 60–70 A. The membrane appeared to be triple layered, each layer having a thickness of 20–25 A.

Teliospore wall structure in Entorrhiza (Tilletiaceae) and its relationship to taxonomy of the genus

1992 ◽  
Vol 70 (10) ◽  
pp. 1964-1983 ◽  
Author(s):  
Brian A. Fineran ◽  
Judith M. Fineran
Keyword(s):  
Electron Microscopy ◽  
Outer Zone ◽  
Spore Wall ◽  
Wall Structure ◽  
Dense Matrix ◽  
Thin Sections ◽  
Transmission Electron ◽  
Layer 2 ◽  
Layer 4 ◽  
Freeze Etching

Spore wall organization in the five species of Entorrhiza (Ustilaginales) has been investigated using thin sections for transmission electron microscopy, supported by light and scanning electron microscopy and some freeze-etching. Material was examined from herbaria, specimens preserved in fixative, and fresh host tissue. The wall has four main layers, numbered 1–4 from the outside to inside of the wall; some layers are further differentiated into zones. Layer 1 in E. aschersoniana, E. caspaiyana, and E. caricicola has two zones: a broad outer zone 2 of dense matrix and a narrow inner zone 1 of less compacted material. Zone 1 is absent in E. cypericola. In E. scirpicola, layer 1 is represented by discontinuous longitudinal ridges. In all spores, layer 2 is composed of a homogeneous electron-dense matrix. Layer 1 in E. aschersoniana, E. casparyana, and E. caricicola is uniformly thick, but in E. cypericola it is broad with an irregular outer margin. In E. scirpicola, layer 2 is differentiated into a distinctive pattern of longitudinal ribs. In all spores of Entorrhiza, layer 3 is resolvable into fine lamellae, corresponding to the mosaic of striations seen after freeze-etching. Layer 3 in Entorrhiza is equivalent to the partition layer described in other Tilletiaceae. Layer 4 has the same organization in all the species, consisting of a very narrow zone 2 abutting layer 3 and a broad zone 1 that forms the rest of the layer. Based on wall structure, E. aschersoniana and E. casparyana represent the most closely related species, followed by E. caricicola, with E. cypericola more distant again. Entorrhiza scirpicola is considered the least related of the species; only its layers 3 and 4 resemble the other species. Key words: Entorrhiza, Tilletiaceae, spore wall ultrastructure, species relationships.

Ultrastructure of Staphylococcus epidermidis after freeze-etching and thin sectioning

1973 ◽  
Vol 19 (2) ◽  
pp. 294-295
Author(s):  
James E. Gilchrist ◽  
Irving W. DeVoe
Keyword(s):  
Staphylococcus Aureus ◽  
Outer Layer ◽  
Staphylococcus Epidermidis ◽  
Cell Walls ◽  
Cytoplasmic Membrane ◽  
Middle Layer ◽  
Dense Layer ◽  
Thin Sections ◽  
Thin Sectioning ◽  
Freeze Etching

A considerable quantity of information is now available on the ultrastructure of Staphylococcus (1, 2, 4, 7, 8, 10, 11, 12). Cell walls of these organisms in thin sections have been shown to consist of three layers: a dense outer layer, a rather electron translucent middle layer, and a very dense layer next to the cytoplasmic membrane (2, 7, 11, 12). Bulger and Bulger (2) have pointed out the presence of circumferential substructure in the middle layer of the wall in a strain of Staphylococcus aureus isolated as the causative agent in fatal pneumonia.Numerous mesosomes of both the vesicular and laminar types are evident in thin sections of staphylococci from several studies (1, 4, 7, 11). Moreover, single vesicular structures that appear to be invaginations of the trilaminar cytoplasmic membrane have been pointed out by Suganuma (11) and Beaton (1).

Fine structure of selected species of the genus Thiobacillus as revealed by chemical fixation and freeze-etching

1974 ◽  
Vol 20 (10) ◽  
pp. 1347-1351 ◽  
Author(s):  
Stanley C. Holt ◽  
J. M. Shively ◽  
J. W. Greenawalt
Keyword(s):  
Inclusion Bodies ◽  
Convex Surface ◽  
Cell Envelope ◽  
Electron Microscopic Examination ◽  
Gram Negative Bacteria ◽  
Chemical Fixation ◽  
Electron Microscopic ◽  
Gram Negative ◽  
Freeze Etching ◽  
Outer Plasma Membrane

An electron-microscopic examination of selected species of the genus Thiobacillus was undertaken using the techniques of chemical fixation and freeze-etching. The architecture of the cells was typical of gram-negative bacteria. The multilayered cell envelope was revealed as a complex of smooth, rough, and particle-studded membranes. The particles which covered the outer plasma membrane (convex surface) appeared to contain a differentiated region which might permit a channeling between the exterior and interior of the cell. Inclusion bodies, including paracrystalline arrays, carboxysomes, and granules were present.

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