geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Use of colloidal gold and ruthenium red in stereo high-voltage electron microscopic study of Con A-binding sites in mouse macrophages


, : Use of colloidal gold and ruthenium red in stereo high-voltage electron microscopic study of Con A-binding sites in mouse macrophages. Histochemistry 81(5): 441-444

Concanavalin A (Con A)-binding sites were labeled with colloidal gold (CG), stained with ruthenium red, and observed under a high-voltage electron microscope. Mouse peritoneal macrophages were labeled by the indirect Con A/CG labeling method at 0 degree C. After washing, some of the cells were incubated in phosphate-buffered saline (PBS) at 37 degrees C. The specimens were then stained with ruthenium red, to enhance the contrast of the cell surface, and embedded in Epon. Sections (0.3 approximately 3 micron thick) were cut and examined by high-voltage electron microscopy at accelerating voltages of 200 approximately 1,000 kV. Staining with ruthenium red provided a strong contrast of the cell surface and the invaginating tubules beneath it against the cytoplasm; in thick sections, both of them were clearly seen by stereomicroscopy. CG particles which represented Con A-binding sites were also sufficiently electron dense to be recognized by high-voltage electron microscopy of thick sections. The two- and three-dimensional distribution of CG particles on the ruthenium-red-positive cell surface was clearly visualized. At 0 degree C, Con A-binding sites were randomly distributed on the cell surface. The redistribution and endocytosis of Con A-binding sites were seen at 37 degrees C. The three-dimensional organization of membrane invagination, which represented the process of endocytosis, was clearly seen by stereomicroscopy. The combination of CG labeling and ruthenium red staining is a useful method for high-voltage electron microscopic analysis of the two- and three-dimensional distribution of CG-labeled ligands on the cell surface in thick sections.

US$29.90

PMID: 6083996


Other references

Adams, C.A.; Xu, D.Z.; Lu, Q.; Deitch, E.A., 2001: Factors larger than 100 kd in post-hemorrhagic shock mesenteric lymph are toxic for endothelial cells. Background. Post-shock mesenteric lymph kills and injures endothelial cells (ECs), but neither the mechanism nor the mediators of lymph's toxic effect are known. Thus, in these studies we investigated and characterized potential factors that...

Okado-Matsumoto, A.; Matsumoto, A.; Fujii, J.; Taniguchi, N., 2001: Effect of cAMP on inducible nitric oxide synthase gene expression: its dual and cell-specific functions. The effects of some cAMP-elevating agents on the induction of nitric oxide synthase II (NOS II) were investigated for a macrophage-derived cell line, RAW264.7, stimulated with lipopolysaccharide (LPS) or interferon-gamma (IFN-gamma) and the result...

Stephen, J.M.; Aine, C.J.; Ranken, D.; Hudson, D.; Shih, J.J., 2003: Multidipole analysis of simulated epileptic spikes with real background activity. This simulated magnetoencephalographic study was designed to determine the variability in source parameters with real subject background activity when applying multidipole spatial-temporal dipole analyses, for which the correct model was compared...

Pratolongo, U., 1943: Biochemical researches on ensilages. According to their biochemical characteristics, one can classify ensilages in 2 classes: (1) autolytic ensilages (fermentations essentially produced by tissue enzymes, which are present in the green fodder before its packing in silos; with great l...

Reitter., 1870: Batrisus schwabii Reitter B. E. Z. xiv p. 212 t. i f. 7, Paskau and Spandau, Ktz.. Batrisus schwabii Reitter B. E. Z. xiv p. 212 t. i f. 7, Paskau (and Spandau, Ktz.)

More, P.; Pai, K., 2012: In vitro NADH-oxidase, NADPH-oxidase and myeloperoxidase activity of macrophages after Tinospora cordifolia (guduchi) treatment. It is believed that the enhanced microbicidal and tumoricidal capability of activated macrophages is related to the remarkable increase in the production of oxygen metabolites. Both the production of H2O2 and the oxidation of NAD(P)H are directly...

Silva J.; Monasterio M.; Sarmiento G., 1971: Ecological survey of the venezuelan western llanos part 2 the northern part of barinas state. Acta Cientifica Venezolana 22(2): 60-71

Aujezdský, Z.; Bríza, J.; Kalvach, J., 1984: Secreting mammary glands in breast carcinoma. Sbornik Lekarsky 86(2-3): 86-88

Lehr; Chatzidimitriou-Dreismann, 1995: General properties of the spectrum of complex scaled Hamiltonians: The transition to two dimensions. Physical Review. A, Atomic, Molecular, and Optical Physics 52(4): 2935-2942

Ebert, E.C.; Roberts, A.I., 2001: The lack of TGFb in the lamina propria contributes to the high levels of IFNg. Gastroenterology 120(5 Supplement 1): A 318, April