Post embedding immunoelectron microscopy of human breast cancer: a comparison of three acrylic resins

Corcoran, David; Walker, R. A.

doi:10.1007/bf01387184

Cited by 4 publications

(3 citation statements)

References 20 publications

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“…LR gold, an aromatic cross-linked acrylic resin, has been successfully employed for immunogold cytochemical localization of various antigenic determinants (Causton 1984;Newman and Hobot 1987;Berryman and Rodewald 1990;Corcoran and Walker 1990;Shires et al 1990;Stirling 1990;Kasper and Migheli 1993;Bienz and Egger 1995), including lysozyme (McPhail et al 1987;Staneva-Dobrovski and Zanger 1995;Zanger 1995). The good handling and cutting characteristics of LR gold, as commended by these authors, were confirmed in the present study.…”

Section: Discussionsupporting

confidence: 80%

Lysozyme expression in the rat parotid gland: light and electron microscopic immunogold studies

Staneva-Dobrovski

1997

Histochemistry and Cell Biology

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Lysozyme (muramidase) is capable of direct bacteriolytic action by hydrolyzing glycosidic bonds in bacterial cell walls. Although it is broadly distributed in vertebrate tissues and secretions, the cellular and subcellular localizations of the enzyme are still not well known. The present study examines the distribution of lysozyme expression in the various cell types of LR gold-embedded rat parotid gland, applying a postembedding immunogold-silver staining technique for light microscopy. Simultaneously, a postembedding immunogold method for electron microscopy was used to determine the cellular compartments engaged in the biosynthesis and exocytosis of lysozyme. Silver-amplified immunogold staining for lysozyme demonstrated identical localization in both paraffin and semithin LR-gold sections: in the supranuclear parts of acinar and intercalated duct cells. Staining intensity varied even between adjacent cells. In the electron microscope, immunogold labeling was detected over the cell compartments associated with protein synthesis and exocytosis in acinar and intercalated duct cells. Lysozyme antigenic sites were visible over endoplasmic reticulum and throughout the Golgi apparatus, being intense over the trans-Golgi network, but even stronger in the condensing vacuoles and most prominent over secretory granules in both cell types. The findings provide the first immunocytochemical evidence of the synthesis and secretion of lysozyme in parotid acinar and intercalated duct cells.

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Section: Discussionsupporting

confidence: 80%

Lysozyme expression in the rat parotid gland: light and electron microscopic immunogold studies

Staneva-Dobrovski

1997

Histochemistry and Cell Biology

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“…Two cell-associated carbohydrate mole cules, SSEA-I and the HNK-I epitope were not immunecytochemically demonstrable in LR White-embedded lisue and it remains to be shown whether these 'LR Whimsensitive' antigens can be immunolabelled in brain tissn ■ embedded in other types of resins such as Lowicryls or LR Gold, which proved to preserve immunogenicity of som ■ antigens better than LR White [26,33], If LR White is use I as embedding carrier. UV light-induced polymerization should be preferred to thermal or catalyst curing.…”

Section: Discussionmentioning

confidence: 99%

“…overnight. Both, heat and catalyst curing may have deleterious effects on the structure of nervous but also of other tissues, such as human prostate [25] and breast cancer [26], rat thyroid gland [27], pancreas [28 , 29], and testes [30]. Poor tissue preservation in these instances may have occurred possibly because the resin polymerized too rapidly, which can cause considerable external contraction of the blocks and disrup tive tissue swelling inside [20,29].…”

Section: Discussionmentioning

confidence: 99%

Use of LR White Resin for Post-Embedding Immunolabelling of Brain Tissue

Gocht¹

1992

Cells Tissues Organs

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The object of this study was to investigate the applicability of the acrylic resin ‘LR White’ to immunolabelling of various antigenic determinants in aldehyde-fixed rat CNS tissue. Antibodies were used, which worked well in paraffin sections and therefore were suitable to detect antigens resistant to complete dehydration and heat. Different LR White embedding protocols were employed in order to select the preparation conditions that adequately preserved both the antigenicity and fine structure. Specimens were completely dehydrated with up to 100% ethanol, which was followed by various infiltration times with LR White monomer. Polymerization of the resin was induced by heat, a chemical catalytic procedure (accelerator), or ultraviolet (UV) light. Paraffin, as well as semithin and ultrathin LR White sections were incubated with antibodies reacting to antigens located on the cell surface (stage-specific embryonic antigen-1; SSEA-1), within the plasma membrane (myelin basic protein), in the cytosol (HNK-1, S100 protein), in the cytoskeleton (GFAP, vimentin, neurofilament protein, INT-FIL), and in the extracellular matrix (laminin). All of the examined antigens were immunocytochemically detectable in paraffin-embedded material, while the carbohydrate moieties, HNK-1 and SSEA-1, were not immunoreactive in LR White sections. However, in cryostat sections processed for pre-embedding immunoelectron microscopy, the HNK-1 epitope and SSEA-1 were immunolabelled. Polymerization carried out under UV light led to better structural preservation of brain tissue than resin cured with heat or catalyst. The length of prior infiltration with monomer apparently had no effect on tissue preservation. Consequently, UV light-induced polymerization of LR White gives acceptable morphology of brain tissue. However, the use of this acrylic resin is restricted to the detection of some CNS antigens only.

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In vitro modulation of cellular localization of milk fat globule membrane antigens in human breast carcinomas

Corcoran

Jones

Walker

1991

The Journal of Pathology

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Alterations in the cellular localization of cell surface components such as the milk fat globule membrane are a common feature of breast carcinomas and relate to the differentiation of a tumour. This study has examined the potential modulation of such components. A group of carcinomas were cultured with and without insulin and/or hydrocortisone and the site of staining for milk fat globule membrane, as detected by the antibodies HMFG 1, HMFG 2, and NCRC 11, was assessed using light microscopic and electron microscopic immunohistochemistry. Modulation of localization, with a shift from cytoplasmic vesicle labelling to submembraneous vesicles/cell surface labelling and intracytoplasmic luminal labelling, was observed in 5 of 14 moderately differentiated and 8 of 11 poorly differentiated carcinomas. Two well differentiated carcinomas continued to show peripheral labelling; three poorly differentiated carcinomas showed no change from cytoplasmic labelling only; and the other carcinomas exhibited heterogeneous localization, making any change difficult to assess. Insulin was required for any change to be observed and it is suggested that this has an effect on the mechanisms for intracellular transport of membrane and secretory proteins.

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Post embedding immunoelectron microscopy of human breast cancer: a comparison of three acrylic resins

Cited by 4 publications

References 20 publications

Lysozyme expression in the rat parotid gland: light and electron microscopic immunogold studies

Lysozyme expression in the rat parotid gland: light and electron microscopic immunogold studies

Use of LR White Resin for Post-Embedding Immunolabelling of Brain Tissue

In vitro modulation of cellular localization of milk fat globule membrane antigens in human breast carcinomas

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