An Embedding Medium Buttress Method to Precisely Position Complexly Contoured Mohs Tissue

2019 ◽  
Vol 45 (6) ◽  
pp. 852-854
Author(s):  
Bryan T. Carroll ◽  
Angel R. Maden
Keyword(s):  
Embedding Medium

Albumins as Embedding Media for Electron Microscopy

1969 ◽  
Vol 27 ◽  
pp. 422-423 ◽  
Author(s):  
J. L. Farrant ◽  
J. D. McLean
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
High Temperature ◽  
Biological Material ◽  
Globular Proteins ◽  
The Past ◽  
Antibody Staining ◽  
Thin Sectioning ◽  
Embedding Medium

For electron microscope techniques such as ferritin-labeled antibody staining it would be advantageous to have available a simple means of thin sectioning biological material without subjecting it to lipid solvents, impregnation with plastic monomers and their subsequent polymerization. With this aim in view we have re-examined the use of protein as an embedding medium. Gelatin which has been used in the past is not very satisfactory both because of its fibrous nature and the high temperature necessary to keep its solutions fluid. We have found that globular proteins such as the serum and egg albumins can be cross-linked so as to yield blocks which are suitable for ultrathin sectioning.

Effects of Vibration on the Preparation of Ultrathin Sections

1973 ◽  
Vol 31 ◽  
pp. 358-359
Author(s):  
Joseph M. Blum ◽  
Edward P. Gargiulo ◽  
J. R. Sawers
Keyword(s):  
Cutting Speed ◽  
Ground Level ◽  
Environmental Vibration ◽  
Block Face ◽  
Ultrathin Sections ◽  
Embedding Medium ◽  
Thickness Variations ◽  
Building Walls ◽  
Cutting Direction ◽  
Diamond Knife

It is now well-known that chatter (Figure 1) is caused by vibration between the microtome arm and the diamond knife. It is usually observed as a cyclical variation in “optical” density of an electron micrograph due to sample thickness variations perpendicular to the cutting direction. This vibration might be induced by using too large a block face, too large a clearance angle, excessive cutting speed, non-uniform embedding medium or microtome vibration. Another prominent cause is environmental vibration caused by inadequate building construction. Microtomes should be installed on firm, solid floors. The best floors are thick, ground-level concrete pads poured over a sand bed and isolated from the building walls. Even when these precautions are followed, we recommend an additional isolation pad placed on the top of a sturdy table.

Cell Fine Structure as Revealed in Dessicator-Dried Resinless Sections

1981 ◽  
Vol 39 ◽  
pp. 622-623
Author(s):  
R. P. Becker ◽  
J. J. Wolosewick ◽  
J. Ross-Stanton
Keyword(s):  
Fine Structure ◽  
Tannic Acid ◽  
Three Dimensional ◽  
Albino Rats ◽  
Cell Organelles ◽  
Vascular Perfusion ◽  
Thin Sections ◽  
Carbon Coated ◽  
Embedding Medium ◽  
Polyethylene Glycol 6000

Methodology has been introduced recently which allows transmission and scanning electron microscopy of cell fine structure in semi-thin sections unencumbered by an embedding medium. Images obtained from these “resinless” sections show a three-dimensional lattice of microtrabeculfee contiguous with cytoskeletal structures and membrane-bounded cell organelles. Visualization of these structures, especially of the matiiDra-nous components, can be facilitated by employing tannic acid in the fixation step and dessicator drying, as reported here.Albino rats were fixed by vascular perfusion with 2% glutaraldehyde or 1.5% depolymerized paraformaldehyde plus 2.5% glutaraldehyde in 0.1M sodium cacodylate (pH 7.4). Tissues were removed and minced in the fixative and stored overnight in fixative containing 4% tannic acid. The tissues were rinsed in buffer (0.2M cacodylate), exposed to 1% buffered osmium tetroxide, dehydrated in ethyl alcohol, and embedded in pure polyethylene glycol-6000 (PEG). Sections were cut on glass knives with a Sorvall MT-1 microtome and mounted onto poly-L-lysine, formvar-carbon coated grids while submerged in a solution of 95% ethanol containing 5% PEG.

A Study of the Ultrastructure of Visual Cell Outer Segment Membranes, Using a Water-Miscible Embedding Medium and Differential Staining

1972 ◽  
Vol 30 ◽  
pp. 50-51
Author(s):  
Anthony J. Godfrey
Keyword(s):  
Outer Segment ◽  
Protein Denaturation ◽  
Uranyl Acetate ◽  
Differential Staining ◽  
Visual Cell ◽  
Lead Citrate ◽  
Chick Retina ◽  
Dark Staining ◽  
Embedding Medium ◽  
The Individual

Aldehyde-fixed chick retina was embedded in a water-containing resin of glutaraldehyde and urea, without dehydration. The loss of lipids and other soluble tissue components, which is severe in routine methods involving dehydration, was thereby minimized. Osmium tetroxide post-fixation was not used, lessening the amount of protein denaturation which occurred. Ultrathin sections were stained with 1, uranyl acetate and lead citrate, 2, silicotungstic acid, or 3, osmium vapor, prior to electron microscope examination of visual cell outer segment ultrastructure, at magnifications up to 800,000.Sections stained with uranyl acetate and lead citrate (Fig. 1) showed that the individual disc membranes consisted of a central lipid core about 78Å thick in which dark-staining 40Å masses appeared to be embedded from either side.

The Application of Ultra-Thin Sectioning Techniques to Non-Biological Samples

1968 ◽  
Vol 26 ◽  
pp. 332-333
Author(s):  
C. F. Oster
Keyword(s):  
Biological Sciences ◽  
Epoxy Resin ◽  
Cross Sections ◽  
Biological Samples ◽  
Industrial Applications ◽  
Photographic Film ◽  
Silver Particles ◽  
Thin Sectioning ◽  
Embedding Medium

Although ultra-thin sectioning techniques are widely used in the biological sciences, their applications are somewhat less popular but very useful in industrial applications. This presentation will review several specific applications where ultra-thin sectioning techniques have proven invaluable.The preparation of samples for sectioning usually involves embedding in an epoxy resin. Araldite 6005 Resin and Hardener are mixed so that the hardness of the embedding medium matches that of the sample to reduce any distortion of the sample during the sectioning process. No dehydration series are needed to prepare our usual samples for embedding, but some types require hardening and staining steps. The embedded samples are sectioned with either a prototype of a Porter-Blum Microtome or an LKB Ultrotome III. Both instruments are equipped with diamond knives.In the study of photographic film, the distribution of the developed silver particles through the layer is important to the image tone and/or scattering power. Also, the morphology of the developed silver is an important factor, and cross sections will show this structure.

Image quality vs data obtainment in biological electron microscopy

1986 ◽  
Vol 44 ◽  
pp. 42-43
Author(s):  
J. E. Johnson
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Electron Beam ◽  
Image Quality ◽  
High Speed ◽  
Early Period ◽  
Early Years ◽  
Fluorescent Screen ◽  
Embedding Medium

In the early years of biological electron microscopy, scientists had their hands full attempting to describe the cellular microcosm that was suddenly before them on the fluorescent screen. Mitochondria, Golgi, endoplasmic reticulum, and other myriad organelles were being examined, micrographed, and documented in the literature. A major problem of that early period was the development of methods to cut sections thin enough to study under the electron beam. A microtome designed in 1943 moved the specimen toward a rotary “Cyclone” knife revolving at 12,500 RPM, or 1000 times as fast as an ordinary microtome. It was claimed that no embedding medium was necessary or that soft embedding media could be used. Collecting the sections thus cut sounded a little precarious: “The 0.1 micron sections cut with the high speed knife fly out at a tangent and are dispersed in the air. They may be collected... on... screens held near the knife“.

The promises and perfidy of cryomicroscopy and analysis

1994 ◽  
Vol 52 ◽  
pp. 382-383
Author(s):  
Patrick Echlin
Keyword(s):  
Low Temperature ◽  
High Energy ◽  
Short Paper ◽  
List Type ◽  
Time Resolved ◽  
Energy Beam ◽  
Cellular Analysis ◽  
Dissolved Ions ◽  
Embedding Medium ◽  
Low Temperature Microscopy

The unusual title of this short paper and its accompanying tutorial is deliberate, because the intent is to investigate the effectiveness of low temperature microscopy and analysis as one of the more significant elements of the less interventionist procedures we can use to prepare, examine and analyse hydrated and organic materials in high energy beam instruments. The promises offered by all these procedures are well rehearsed and the litany of petitions and responses may be enunciated in the following mantra.Vitrified water can form the perfect embedding medium for bio-organic samples.Frozen samples provide an important, but not exclusive, milieu for the in situ sub-cellular analysis of the dissolved ions and electrolytes whose activities are central to living processes.The rapid conversion of liquids to solids provides a means of arresting dynamic processes and permits resolution of the time resolved interactions between water and suspended and dissolved materials.The low temperature environment necessary for cryomicroscopy and analysis, diminish, but alas do not prevent, the deleterious side effects of ionizing radiation.Sample contamination is virtually eliminated.

scholarly journals Improvements of glycol methacrylate. I. Its use as an embedding medium for electron microscopic studies.

1977 ◽  
Vol 25 (3) ◽  
pp. 163-174 ◽  
Author(s):  
R C Spaur ◽  
G C Moriarty
Keyword(s):  
Electron Beam ◽  
Water Soluble ◽  
Cross Linking ◽  
Electron Microscopic ◽  
Glycol Methacrylate ◽  
Ultrastructural Studies ◽  
Tissue Morphology ◽  
Methacrylate Monomer ◽  
Microscopic Studies ◽  
Embedding Medium

The technique for using the water-soluble embedding medium glycol methacrylate has been improved for ultrastructural studies by the simplification of the method of formation of prepolymers used in embedding the tissue, by the addition of a cross-linking agent so that sections are stable in the electron beam, and by improving the softness of the blocks by the addition of a plasticizing agent. The preservation of tissue morphology has been improved by complete dehydration in glycol methacrylate monomer prior to infiltration with the prepolymer. Preservations of tissue morphology is further enhanced by complete dehydration in ethanols and embedding in the improved glycol methacrylate medium.

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