An epoxy resin and silicone impregnation technique for the preservation of oral pathology teaching specimens

This paper is dedicated to an impregnation technique for crack identification of uniaxial tensile behaviour of concrete samples. A method for crack identification in concrete after uniaxial tension tests was adopted for the observation of differences between various experiments which were conducted with and without the elimination of secondary flexure. During the procedure an epoxy resin containing fluorescent dye was infused into concrete samples by vacuum to expose cracks and defects. After impregnation the samples were sawed from the prism and pictures taken under ultraviolet light.

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Ultrastructural investigation of spontaneous pituitary gonadotroph cell adenomas in aging Sprague-Dawley rats

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100077086 ◽

1983 ◽

Vol 41 ◽

pp. 702-703

Author(s):

D. J. McComb ◽

J. Beri ◽

F. Zak ◽

K. Kovacs

Keyword(s):

Electron Microscopy ◽

Animal Model ◽

Epoxy Resin ◽

Immunoelectron Microscopy ◽

Tumor Type ◽

Gonadal Function ◽

Sprague Dawley ◽

Male And Female ◽

Reticulin Fibers ◽

Sprague Dawley Rats

Gonadotroph cell adenomas of the pituitary are infrequent in human patients and are not invariably associated with altered gonadal function. To date, no animal model of this tumor type exists. Herein, we describe spontaneous gonadotroph cell adenomas in old male and female Sprague-Dawley rats by histology, immunocytology and electron microscopy.The material consisted of the pituitaries of 27 male and 38 female Sprague Dawley rats, all 26 months of age or older, removed at routine autopsy. Sections of formal in-fixed, paraffin-embedded tissue were stained with hematoxylin-phloxine-saffron (HPS), the PAS method and the Gordon-Sweet technique for the demonstration of reticulin fibers. For immunostaining, sections were exposed to anti-rat β-LH, anti-ratβ-TSH, anti-rat PRL, anti-rat GH and anti-rat ACTH 1-39. For electron microscopy, tissue was fixed in 2.5% glutaraldehyde, postfixed in 1% OsO4 and embedded in epoxy-resin. Tissue fixed in 10% formalin, embedded in epoxy resin without osmification, was used for immunoelectron microscopy.

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Surface Structure in Microtomed Sections Caused by Glass and Diamond Knives

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066334 ◽

1971 ◽

Vol 29 ◽

pp. 456-457

Author(s):

J. Temple Black ◽

William G. Boldosser

Keyword(s):

Plastic Deformation ◽

Epoxy Resin ◽

Carbon Coating ◽

Surface Damage ◽

Body Angle ◽

Normal Manner ◽

Bottom Side ◽

Cutting Direction ◽

Made In ◽

Diamond Knife

Ultramicrotomy produces plastic deformation in the surfaces of microtomed TEM specimens which can not generally be observed unless special preparations are made. In this study, a typical biological composite of tissue (infundibular thoracic attachment) infiltrated in the normal manner with an embedding epoxy resin (Epon 812 in a 60/40 mixture) was microtomed with glass and diamond knives, both with 45 degree body angle. Sectioning was done in Portor Blum Mt-2 and Mt-1 microtomes. Sections were collected on formvar coated grids so that both the top side and the bottom side of the sections could be examined. Sections were then placed in a vacuum evaporator and self-shadowed with carbon. Some were chromium shadowed at a 30 degree angle. The sections were then examined in a Phillips 300 TEM at 60kv.Carbon coating (C) or carbon coating with chrom shadowing (C-Ch) makes in effect, single stage replicas of the surfaces of the sections and thus allows the damage in the surfaces to be observable in the TEM. Figure 1 (see key to figures) shows the bottom side of a diamond knife section, carbon self-shadowed and chrom shadowed perpendicular to the cutting direction. Very fine knife marks and surface damage can be observed.

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Epoxy Resin Embedment

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010010024x ◽

1968 ◽

Vol 26 ◽

pp. 100-101

Author(s):

J. G. Adams ◽

M. M. Campbell ◽

H. Thomas ◽

J. J. Ghldonl

Keyword(s):

Electron Microscopy ◽

Electron Beam ◽

Epoxy Resin ◽

Light Microscope ◽

Epoxy Resins ◽

Image Contrast ◽

Tissue Preservation ◽

Resin System ◽

Excellent Quality

Since the introduction of epoxy resins as embedding material for electron microscopy, the list of new formulations and variations of widely accepted mixtures has grown rapidly. Described here is a resin system utilizing Maraglas 655, Dow D.E.R. 732, DDSA, and BDMA, which is a variation of the mixtures of Lockwood and Erlandson. In the development of the mixture, the Maraglas and the Dow resins were tested in 3 different volumetric proportions, 6:4, 7:3, and 8:2. Cutting qualities and characteristics of stability in the electron beam and image contrast were evaluated for these epoxy mixtures with anhydride (DDSA) to epoxy ratios of 0.4, 0.55, and 0.7. Each mixture was polymerized overnight at 60°C with 2% and 3% BDMA.Although the differences among the test resins were slight in terms of cutting ease, general tissue preservation, and stability in the beam, the 7:3 Maraglas to D.E.R. 732 ratio at an anhydride to epoxy ratio of 0.55 polymerized with 3% BDMA proved to be most consistent. The resulting plastic is relatively hard and somewhat brittle which necessitates trimming and facing the block slowly and cautiously to avoid chipping. Sections up to about 2 microns in thickness can be cut and stained with any of several light microscope stains and excellent quality light photomicrographs can be taken of such sections (Fig. 1).

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100101402 ◽

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.

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Ultrastructure of the basal body apparatus in epidermal cells of a sponge larva (Aplysilla SP: Demospongiae)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012504x ◽

1992 ◽

Vol 50 (1) ◽

pp. 928-929

Author(s):

R.L. Pinto ◽

R.M. Woollacott

Keyword(s):

Sodium Bicarbonate ◽

Epoxy Resin ◽

Basal Body ◽

Phosphate Buffer ◽

Similar Data ◽

Basal Apparatus ◽

Striated Rootlet ◽

The Common ◽

Basal Foot ◽

Sexually Mature

The basal body and its associated rootlet are the organelles responsible for anchoring the flagellum or cilium in the cytoplasm. Structurally, the common denominators of the basal apparatus are the basal body, a basal foot from which microtubules or microfilaments emanate, and a striated rootlet. A study of the basal apparatus from cells of the epidermis of a sponge larva was initiated to provide a comparison with similar data on adult sponges.Sexually mature colonies of Aplysillasp were collected from Keehi Lagoon Marina, Honolulu, Hawaii. Larvae were fixed in 2.5% glutaraldehyde and 0.14 M NaCl in 0.2 M Millonig’s phosphate buffer (pH 7.4). Specimens were postfixed in 1% OsO4 in 1.25% sodium bicarbonate (pH 7.2) and embedded in epoxy resin. The larva ofAplysilla sp was previously described (as Dendrilla cactus) based on live observations and SEM by Woollacott and Hadfield.

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A rapid silver-impregnation technique on ultra-thin sections for Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100147053 ◽

1993 ◽

Vol 51 ◽

pp. 242-243

Author(s):

K. Chien ◽

R.C. Heusser ◽

M.L. Jones ◽

R.L. Van de Velde

Keyword(s):

Electron Microscopy ◽

Silver Nitrate ◽

Silver Impregnation ◽

Sodium Borate ◽

Renal Diseases ◽

Distilled Water ◽

Thin Sections ◽

Impregnation Technique ◽

Centrifuge Tube ◽

Simplified Procedure

Silver impregnation techniques have been used for the demonstration of the complex carbohydrates in electron microscopy. However, the silver stains were believed to be technically sensitive and time consumming to perform. Currently, due to the need to more specifically evaluate immune complex for localization in certain renal diseases, a simplified procedure in conjunction with the use of the microwave has been developed and applied to renal and other biopsies. The procedure is as follows:Preparation of silver methenamine solution:1. 15ml graduated, clear polystyrene centrifuge tube (Falcon, No. 2099) was rinsed once with distilled water.2. 3% hexamethylene tetramine (methenamine) was added into the centrifuge tube to the 6ml mark.3. 3% silver nitrate was added slowly to the methenamine to the 7ml mark while agitating. (Solution will instantly turn milky in color and then clear rapidly by mixing. No precipitate should be formed).4. 2% sodium borate was added to the solution to the 8ml mark, mixed and centrifuged before use.

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