Selective Staining of Acid Mucopolysaccharides By Ruthenium Red

1968 ◽  
Vol 26 ◽  
pp. 38-39
Author(s):  
J. H. Luft
Keyword(s):  
Electron Microscope ◽  
Light Microscopy ◽  
Light Microscope ◽  
Osmium Tetroxide ◽  
Single Cells ◽  
Ruthenium Red ◽  
Collagen Fibers ◽  
Selective Staining ◽  
Pectic Substances ◽  
Solid Tissues

Ruthenium red is one of the few completely inorganic dyes used to stain tissues for light microscopy. This novelty is enhanced by ignorance regarding its staining mechanism. However, its continued usefulness in botany for demonstrating pectic substances attests to selectivity of some sort. Whether understood or not, histochemists continue to be grateful for small favors.Ruthenium red can also be used with the electron microscope. If single cells are exposed to ruthenium red solution, sufficient mass can be bound to produce observable density in the electron microscope. Generally, this effect is not useful with solid tissues because the contrast is wasted on the damaged cells at the block surface, with little dye diffusing more than 25-50 μ into the interior. Although these traces of ruthenium red which penetrate between and around cells are visible in the light microscope, they produce negligible contrast in the electron microscope. However, its presence can be amplified by a reaction with osmium tetroxide, probably catalytically, to be easily visible by EM. Now the density is clearly seen to be extracellular and closely associated with collagen fibers (Fig. 1).

scholarly journals THE FINE STRUCTURE OF THE RAT CEREBELLUM

1964 ◽  
Vol 23 (2) ◽  
pp. 277-293 ◽  
Author(s):  
Robert M. Herndon
Keyword(s):  
Fine Structure ◽  
Electron Microscope ◽  
Light Microscope ◽  
Osmium Tetroxide ◽  
Granule Cells ◽  
Cell Types ◽  
Bergmann Glia ◽  
Rat Cerebellum

This paper describes the fine structure of the granule cells, stellate neurons, astrocytes, Bergmann glia, oligodendrocytes, and microglia of the rat cerebellum after fixation by perfusion with buffered 1 per cent osmium tetroxide. Criteria are given for differentiating the various cell types, and the findings are correlated with previous light microscope and electron microscope studies of the cerebellum.

Comparison of Larval Stored Product Beetle Mandibles by Light Microscope and Scanning Electron Microscope

1981 ◽  
Vol 64 (1) ◽  
pp. 199-224
Author(s):  
John E Kvenberg
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Light Microscopy ◽  
Light Microscope ◽  
Morphological Characteristics ◽  
Scanning Electron ◽  
Conventional Light Microscopy

Abstract Larval stored product beetle mandibles were studied by comparing images made by scanning electron microscopy with those made by conventional light microscopy. Discussion of morphological characteristics is based on illustrations of 25 species

Ruthenium Red as a Ligand of Osmium for Examining Uncoated Biological Material in the Scanning Electron Microscope

1977 ◽  
Vol 35 ◽  
pp. 554-555
Author(s):  
Alfredo Feria-Velasco ◽  
Jorge Arauz-Contreras
Keyword(s):  
Electron Microscope ◽  
High Resolution ◽  
Scanning Electron Microscope ◽  
Biological Material ◽  
Carbon Coating ◽  
Osmium Tetroxide ◽  
Metal Layer ◽  
Ruthenium Red ◽  
Gold Coating ◽  
Scanning Electron

The scanning electron microscope (SEM) has great advantages over the light microscope for examining the surface of cells and tissues, and recently it has been employed for studying the distribution of enzymes and antigens on their surfaces (1). In the basic methodology for preparing specimens to be examined in the SEM, the samples are coated with a 20-40 nm metal layer, usually gold, platinum and palladium. For high resolution SEM purposes, the thinnest possible conducting coat should be used and some authors have employed a 2-5 nm gold coating, omiting the preliminary carbon coating (2). However, some workers using the evaporative metal techniques have encountered difficulties and possible artifacts even with the thinnest evaporated metal layer (3). Various authors have described procedures for examining uncoated specimens, being one of the most useful methods the modified thiocarbohydrazide procedure for SEM (4). In the present work a method for examining the uncoated specimens in the SEM was used by means of osmium tetroxide (OSO4) and employing ruthenium red (RR) as a ligand.

Ultrastructure of Angiosarcoma of Mouse Tail

1980 ◽  
Vol 38 ◽  
pp. 740-741
Author(s):  
White Yvonne ◽  
Winslow Sheldon ◽  
James W. Townsend ◽  
Neil A. Littlefield
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Light Microscopy ◽  
Toluidine Blue ◽  
Female Mouse ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Ultrathin Sections ◽  
Resin Mixture

Spontaneous neoplasms rarely occur on the tails of BALB/cStCrlfC3H/Nctr mice, but the neoplasm most frequently observed is a locally invasive non-metastatic angiosarcoma. In this case a female mouse weighing 33.2 g, 706 days of age, presented a soft, red, irregularly-shaped mass, measuring 18 mm in its greatest dimension, in the subcutis of the base of the tail. A portion of the tail tumor was taken for electron microscopy and the remainder was processed for light microscopy. The tissue processed for electron microscopy was fixed in 4% cacodyl ate-buffered glutaraldehyde, post-fixed in 1% osmium tetroxide, dehydrated in a graded series of ethanol solutions, and embedded in Epon-Araldite resin mixture. Sections of 1 μm were stained with toluidine blue for light microscopy and ultrathin sections of 100 nm were stained with uranyl acetate and lead citrate, then examined with a Philips EM201 electron microscope .

A Simple Light-Microscope Technique for the Examination of Cell Surfaces

1973 ◽  
Vol 13 (1) ◽  
pp. 317-319
Author(s):  
A. ROSS ◽  
I. P. GORMLEY
Keyword(s):  
Heavy Metal ◽  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Light Microscopy ◽  
Light Microscope ◽  
Simple Technique ◽  
Cell Surfaces ◽  
Microscope Technique ◽  
Scanning Electron

A simple technique for the examination of cell surfaces by shadowing them with a heavy metal and examining by light microscopy has been described. Stained preparations can be examined and photographed initially prior to investigation by this technique. It is suggested that this technique could be used instead of the scanning electron microscope or replica techniques where the limited resolution of the light microscope is sufficient.

Lipid staining for the electron microscope: a new method

1975 ◽  
Vol 19 (3) ◽  
pp. 425-437
Author(s):  
V.B. Wigglesworth
Keyword(s):  
Electron Microscope ◽  
Light Microscope ◽  
Osmium Tetroxide ◽  
New Method ◽  
Monoterpene Hydrocarbon ◽  
Ethyl Gallate ◽  
Unsaturated Lipids ◽  
Specific Increase

Tissues fixed in osmium tetroxide or in combined osmium and glutaraldehyde (Hinde), embedded in Spurr's medium, cut at 0-5-I mum and mounted in Farrants' gum medium containing ethyl gallate, show good staining of lipid-contaning structures (droplets of triglyceride, membranes, mitochondria, etc.) in the light microscope. Such preparations show moderate contrast in the electron microscope without further staining. But a specific increase in contrast in lipid-rich structures is obtained by partition of the tissues, before embedding, in 70% ethanol saturated with the monoterpene hydrocarbon myrcene, with or without the addition of 0-I % ethyl gallate, followed by osmium tetroxide. This method will visualize both saturated and unsaturated lipids, including waxes.

scholarly journals A CORRELATIVE STUDY BY ELECTRON AND LIGHT MICROSCOPY OF THE DEVELOPMENT OF TYPE 5 ADENOVIRUS

1960 ◽  
Vol 112 (2) ◽  
pp. 383-402 ◽  
Author(s):  
Gabriel C. Godman ◽  
Councilman Morgan ◽  
Peter M. Breitenfeld ◽  
Harry M. Rose
Keyword(s):  
Electron Microscope ◽  
Nucleic Acid ◽  
Light Microscopy ◽  
Hela Cells ◽  
Light Microscope ◽  
Protein Bodies ◽  
Correlative Study ◽  
Electron And Light Microscopy

The evolution of the intranuclear lesion produced by type 5 adenovirus in HEp-2 and HeLa cells is described as seen in the light microscope and the bodies formed in the course of the infection characterized histochemically. Some 12 hours after infection acidophilic protein bodies, without appreciable nucleic acid, first appear in the nucleus and coalesce into a network. Within or in association with this material, DNA-containing masses (viral aggregates) are formed which rapidly increase in amount and then coalesce. At the same time, a protein is produced, histochemically different from that of the acidophilic or basophilic structures mentioned, within the infected nucleus, which constitutes a matrix within which regular cytstals of a protein, (presumably non-viral) materialize. These structural and histochemical features are correlated with details which have been observed in parallel studies with the electron microscope.

scholarly journals "NEW MEMBRANE" FORMATION IN AMOEBA PROTEUS UPON INJURY OF INDIVIDUAL CELLS

1971 ◽  
Vol 49 (3) ◽  
pp. 747-772 ◽  
Author(s):  
Barbara Szubinska
Keyword(s):  
Plasma Membrane ◽  
Electron Microscope ◽  
Single Cells ◽  
Amoeba Proteus ◽  
Ruthenium Red ◽  
Membrane Formation ◽  
Frequent Contact ◽  
Membrane Complex

Changes in the plasma membrane complex following the injury of single cells of Amoeba proteus were examined with the electron microscope. Two types of injury were employed in this study; cells were either pinched ("cut") in half or speared with a glass microneedle, and quickly fixed. Speared cells, when fixed in the presence of the ruthenium violet (a derivative of ruthenium red), revealed the presence of an extra trilaminar structure outside of each cell. This structure, called the "new membrane," was separated from the plasma membrane complex by a distance of less than a micron. The trilaminar structure of the new membrane strikingly resembled the image of the plasma membrane in all cells examined, except for its increased width (30%). This new membrane appeared nearly to surround the injured amebae. Attempts were made to demonstrate the possible origin of the new membrane, its reality, and its sensitivity to calcium. Also, some evidence is shown concerning the role of the small dense droplets (100–1200 A in diameter) normally present in the cytoplasm of amebae. Their frequent contact with the plasma membrane of the cell as the result of injury is interpreted as indicating their involvement in the formation and expansion of the plasma membrane.

A Study of the Kultschitzky (Argentaffin) Cell with the Electron-microscope, after Fixation by Osmium Tetroxide

1955 ◽  
Vol s3-96 (35) ◽  
pp. 295-299
Author(s):  
A. C. CHRISTIE
Keyword(s):  
Electron Microscope ◽  
Guinea Pig ◽  
Light Microscope ◽  
Alimentary Canal ◽  
Osmium Tetroxide ◽  
Maximum Diameter ◽  
Osmium Tetroxide Solution ◽  
Argentaffin Cell

1. The Kultschitzky cells of the alimentary canal of man and the guinea-pig were studied with the light-microscope, buffered osmium tetroxide being used as fixative. 2. The Kultschitzky cells of the guinea-pig were shown by the electron-microscope to contain spheroidal granules having a maximum diameter of 0.3µ. These granules are considerably darkened by fixation for only 4 hours in 1 % osmium tetroxide solution. 3. The opinion that the granules of the Kultschitzky cell are only an artifact of formaldehyde fixation is denied.

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