Simplified procedures for releasing and concentrating microorganisms from soil for transmission electron microscopy viewing as thin-sectioned and frozen-etched preparations

1975 ◽  
Vol 21 (3) ◽  
pp. 252-262 ◽  
Author(s):  
D. L. Balkwill ◽  
D. P. Labeda ◽  
L. E. Casida Jr.
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Types ◽  
Soil Slurry ◽  
Thin Sections ◽  
Choice Of Procedure ◽  
Transmission Electron ◽  
Cell Release ◽  
Simplified Procedures ◽  
Simplified Procedure

A simplified procedure is presented for releasing and concentrating indigenous microbial cells from soil for viewing by transmission electron microscopy as thin sections or replicas of frozen-etched preparations. This procedure is compared with two others reported earlier, and their relative merits are discussed as concerns the choice of procedure for the cellular information desired from the soil. Freeze-etching showed that the cell types and size distributions for cells which have been released and concentrated from soil are in general agreement with those for cells in a crude soil slurry in which no attempt to release and concentrate cells was made. Microcolonies were present both in the crude slurry and in the discard soil debris centrifugation pellets from the cell release and concentration procedures. In contrast to the historic assumptions, these microcolonies, as well as some individual cells embedded in soil debris could not be broken up and (or) dislodged so that they would be washed from the soil. The relative numbers of these cells remaining with the soil debris, however, could not be quantitated in the present study.

Two- or three-way observations of the identical cell elements within the same sections by LM, TEM and/or SEM

1978 ◽  
Vol 36 (2) ◽  
pp. 82-83 ◽  
Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Light Microscopy ◽  
Thin Sections ◽  
Transmission Electron ◽  
Identical Cell ◽  
Electron Microscopes ◽  
Scanning Electron

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).

Dislocations in alumina deformed by prismatic slip

1978 ◽  
Vol 36 (1) ◽  
pp. 606-607
Author(s):  
J. Cadoz ◽  
J. Castaing ◽  
J. Philibert
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Basal Slip ◽  
Prismatic Slip ◽  
Compression Tests ◽  
Thin Sections ◽  
Burgers Vector ◽  
Maximum Deformation ◽  
Transmission Electron ◽  
Mechanical Thinning

Plastic deformation of alumina has been much studied; basal slip occurs and dislocation structures have been investigated by transmission electron microscopy (T.E.M.) (1). Non basal slip has been observed (2); the prismatic glide system <1010> {1210} has been obtained by compression tests between 1400°C and 1800°C (3). Dislocations with <0110> burgers vector were identified using a 100 kV microscope(4).We describe the dislocation structures after prismatic slip, using high voltage T.E.M. which gives much information.Compression tests were performed at constant strainrate (∿10-4s-1); the maximum deformation reached was 0.03. Thin sections were cut from specimens deformed at 1450°C, either parallel to the glide plane or perpendicular to the glide direction. After mechanical thinning, foils were produced by ion bombardment. Details on experimental techniques can be obtained through reference (3).

Dislocations and stacking faults in stainless steel

1957 ◽  
Vol 240 (1223) ◽  
pp. 524-538 ◽  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Stainless Steel ◽  
Grain Boundaries ◽  
Stacking Faults ◽  
Stacking Fault ◽  
Stacking Fault Energy ◽  
Thin Sections ◽  
Partial Dislocations ◽  
Transmission Electron

Further experiments by transmission electron microscopy on thin sections of stainless steel deformed by small amounts have enabled extended dislocations to be observed directly. The arrangement and motion of whole and partial dislocations have been followed in detail. Many of the dislocations are found to have piled up against grain boundaries. Other observations include the formation of wide stacking faults, the interaction of dislocations with twin boundaries, and the formation of dislocations at thin edges of the foils. An estimate is made of the stacking-fault energy from a consideration of the stresses present, and the properties of the dislocations are found to be in agreement with those expected from a metal of low stacking-fault energy.

Neoformation of halloysite on volcanic glass in a marine environment

Clay Minerals ◽  
1987 ◽  
Vol 22 (2) ◽  
pp. 179-185 ◽  
Author(s):  
T. Imbert ◽  
A. Desprairies
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Marine Environment ◽  
Experimental Studies ◽  
Volcanic Glass ◽  
Marine Environments ◽  
Thin Sections ◽  
Transmission Electron ◽  
Glass Shards

AbstractTransmission electron microscopy of ultramicrotomed thin-sections of Pleistocene and Eocene glass shards revealed the neoformation of (i) illite and (ii) halloysite at the glass periphery. According to previous experimental studies, halloysite neoformation in marine environments can occur on glass shards deposited in Si-rich sediments; an excess of Ca tends to inhibit the reaction.

Demountable polished extra-thin sections and their use in transmission electron microscopy

1981 ◽  
Vol 44 (335) ◽  
pp. 357-359 ◽  
Author(s):  
D. J. Barber
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Epoxy Resin ◽  
Essential Feature ◽  
Heterogeneous Materials ◽  
Glass Slide ◽  
Thin Sections ◽  
Fine Grained ◽  
Transmission Electron ◽  
Polished Side

The advantages of polished ultra-thin sections (PUTS) in the study of very fine-grained materials, such as occur in some meteorites, have been illustrated by Fredriksson et al. (1978) whose technique is based on the earlier work of Beauchamp and WiUiford (1974). An essential feature of such methods for friable and heterogeneous materials is the use of a medium, usually an epoxy resin, to consolidate and partially impregnate them. Normally one polished side of the specimen is bonded to a glass slide during preparation, and the finished PUTS are integral with the slide on completion. PUTS are typically 2-5 microns in thickness.

scholarly journals Mitochondria of Human Adrenal Cortex Have Tubular Cristae with Bulbous Tips

2003 ◽  
Vol 88 (4) ◽  
pp. 1903-1906 ◽  
Author(s):  
Alessandro Riva ◽  
Felice Loffredo ◽  
Alessandro Uccheddu ◽  
Francesca Testa Riva ◽  
Bernard Tandler
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
High Resolution ◽  
Adrenal Cortex ◽  
Thin Sections ◽  
Transmission Electron ◽  
Soluble Material ◽  
Bulbous Tip ◽  
Scanning Electron

By taking advantage of a modified osmium maceration technique, we have been able to examine by high resolution scanning electron microscopy (HRSEM) the interior of human adrenocortical mitochondria from which all soluble material has been extracted. The so-called vesicles apparent in thin sections examined by transmission electron microscopy actually are finger-like cristae as determined by HRSEM. These digitiform cristae have a segmented appearance and a bulbous tip. The segmented form of the cristae may have important metabolic implications.

scholarly journals The exit of Trypanosoma cruzi from the phagosome is inhibited by raising the pH of acidic compartments.

1990 ◽  
Vol 171 (2) ◽  
pp. 401-413 ◽  
Author(s):  
V Ley ◽  
E S Robbins ◽  
V Nussenzweig ◽  
N W Andrews
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Trypanosoma Cruzi ◽  
Ammonium Chloride ◽  
Protozoan Parasite ◽  
Cell Types ◽  
Immunocytochemical Localization ◽  
Transmission Electron ◽  
Acidic Compartments ◽  
Vacuolar Ph

The protozoan parasite Trypanosoma cruzi can infect many distinct mammalian cell types. The parasites enter cells through the formation of phagocytic vacuoles, but later are found free in the cytosol, where they multiply as amastigotes. Using transmission electron microscopy we found that within 2 h after infection 70% of the parasites, including examples of both mammalian forms (trypomastigotes and amastigotes), were inside partially disrupted vacuoles or free in the cytosol. We demonstrated that the pH of vacuoles containing recently interiorized parasites is acidic, through immunocytochemical localization of the acidotropic compound DAMP (18) in their interior. Increasing the vacuolar pH with chloroquine, ammonium chloride, methylamine, or monensin significantly inhibited the escape of the parasites into the cytosol. These results are compatible with the hypothesis that an acid-active hemolysin of T. cruzi (15) might be involved in the escape mechanism.

scholarly journals Improved Sectioning of Polymers Using an Oscillating Diamond Knife for Transmission Electron Microscopy

2006 ◽  
Vol 14 (5) ◽  
pp. 20-21 ◽  
Author(s):  
J.D. Harris ◽  
J.S. Vastenhout
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Wedge Angle ◽  
Cutting Speed ◽  
Sample Surface ◽  
Section Thickness ◽  
Thin Sections ◽  
Transmission Electron ◽  
Surface Section ◽  
Diamond Knife

Polymers are viscoelastic materials that can often deform during microtome sectioning. Similar to plastic embedded biological materials, many methods have been developed over the years to not only improve the image contrast of these materials but also to harden the material for improved sectioning during microtomy. Even with these improvements, a common artifact, compression, during the sectioning of this class of materials remains problematic.Compression is caused by several factors: hardness of the sample, embedding media, wedge angle of the knife, interaction between the diamond and sample surface, section thickness and cutting speed. It has been found that reducing the knife angle from 45º to 35° leads to a reduction in compression. Recent efforts to further reduce the compression of ultra-thin sections have led to the invention of an oscillating diamond knife.

scholarly journals Three-Dimensional Scanning Transmission Electron Microscopy of Biological Specimens

2010 ◽  
Vol 16 (1) ◽  
pp. 54-63 ◽  
Author(s):  
Niels de Jonge ◽  
Rachid Sougrat ◽  
Brian M. Northan ◽  
Stephen J. Pennycook
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Mammalian Cells ◽  
Three Dimensional ◽  
Axial Resolution ◽  
Beam Radiation ◽  
Thin Sections ◽  
Scanning Transmission Electron ◽  
Transmission Electron ◽  
Scanning Transmission

AbstractA three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2–3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original dataset. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy. However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved dataset.

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