An Attempt to Measure Glomerular Plasma Flow by the Cationic Dyes Alcian Blue and Ruthenium Red

1985 ◽  
Vol 8 (3) ◽  
pp. 136-139
Author(s):  
O.J. Lavik ◽  
K. Aukland
Keyword(s):  
Alcian Blue ◽  
Plasma Flow ◽  
Ruthenium Red ◽  
Cationic Dyes

scholarly journals Proteoglycans in arterial smooth muscle cell cultures: an ultrastructural histochemical analysis.

1984 ◽  
Vol 32 (4) ◽  
pp. 347-357 ◽  
Author(s):  
K Chen ◽  
T N Wight
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Acridine Orange ◽  
Alcian Blue ◽  
Ruthenium Red ◽  
Cationic Dyes ◽  
Monolayer Films ◽  
Arterial Smooth Muscle ◽  
Chondroitinase Abc ◽  
Safranin O

The extracellular matrix in cultures of arterial smooth muscle cells has been examined by ultrastructural histochemistry using each of the following cationic dyes: ruthenium red, Alcian blue, acridine orange, and safranin O. All dyes exhibited an affinity for a structural component that was either preserved as a granule with ruthenium red or Alcian blue, or as an extended filament or bottlebrush structure with acridine orange or safranin O. Both granules and filaments were removed when the cultures were pretreated with chondroitinase ABC, an enzyme that degrades the glycosaminoglycan moiety of some proteoglycans. These structural components of the extracellular matrix were not observed when cultures were prepared in the absence of the cationic dyes. Labeling experiments (35S-sulfate) revealed that approximately 40% of the total labeled proteoglycans were lost during routine processing for electron microscopy (i.e., fixation through dehydration). Inclusion of any one of the cationic dyes during fixation reduced the losses to less than 1%. The extended filamentous structure preserved by safranin O and acridine orange resembled the structure of purified proteoglycans prepared from the same cultures and spread on cytochrome c monolayer films. These observations suggest that proteoglycans exist as extended bottlebrush structures within the extracellular matrix, and support the interpretation that the granular deposits observed in the ruthenium red and Alcian blue preparations most likely represent individual proteoglycan monomers that have undergone molecular collapse during processing. In addition, the dyes also exhibited an affinity for chords of fine fibrils that contained small granules and/or filaments. Both the fibrillar material and the associated granular and filamentous structures enmeshed in the fibrils resisted digestion with chondroitinase ABC.

Cell surface saccharides of Trypanosoma lewisi. I. Polycation-induced cell agglutination and fine-structure cytochemistry

1975 ◽  
Vol 19 (3) ◽  
pp. 621-644
Author(s):  
D.M. Dwyer
Keyword(s):  
Fine Structure ◽  
Cell Surface ◽  
Alcian Blue ◽  
Chondroitin Sulphate ◽  
Surface Membrane ◽  
Ruthenium Red ◽  
Surface Coat ◽  
Cell Agglutination ◽  
Trypanosoma Lewisi ◽  
Cationic Compounds

Trypanosoma lewisi bloodstream and culture forms were agglutinated differentially with low concentrations of the cationic compounds: ruthenium red, ruthenium violet, Alcian blue chloride, 1-hexadecylpyridinium chloride, lanthanum chloride, and cationized ferritin. The bloodstream form trypanosomes gave the highest agglutination levels with each of the compounds tested. Ruthenium red was the most effective inducer of cell agglutination among the several cations used. Trypsin-treated bloodstream forms were agglutinated less in the presence of ruthenium red than untreated controls. Ruthenium red-induced cell agglutination also was lowered with chondroitin sulphate and dextran sulphate, but not with alpha-D-glucose, alpha-D-mannose or with several methyl glycosides. Treatment of the bloodstream trypanosomes with alpha-amylase, dextranase, or neuraminidase had little effect on agglutination levels obtained with ruthenium red. Fine-structure cytochemical staining with ruthenium red, ruthenium violet, and Alcian blue-lanthanum nitrate was used to ascertain the presence and distribution of presumptive carbohydrates in the trypanosome cell surface. The extracellular surface coat of the bloodstream forms stained densely with each of the polycationic dyes. Trypsin treatment removed the surface coat from bloodstream trypanosomes; however, the surface membranes of the organisms were stained densely with the several dyes. Similar surface-membrane staining was obtained with the cationic compounds and the culture forms, which lack a cell surface coat. Cationized ferrin was used at the fine-structure level to visualize the negative surface charge present in the cell surface coat and external membrane of the several trypanosome stages. Results obrained from the agglutination and cytochemistry experiments indicate that complex polysaccharides are present in the surface membranes and cell surface coat of T. lewisi bloodstream forms. Similar conclusions also pertain to the surface membranes of the T. lewisi culture from trypanosomes. The carbohydrates probably represent glycopeptide and glycoprotein structural components of the surface membrane of this organism.

Compositional and structural heterogenicity of the cardiac jelly of the chick embryo tubular heart: a TEM, SEM and histochemical study

Development ◽  
1980 ◽  
Vol 56 (1) ◽  
pp. 211-223
Author(s):  
J. M. Hurle ◽  
J. M. Icardo ◽  
J. L. Ojeda
Keyword(s):  
Alcian Blue ◽  
Regional Differences ◽  
Histochemical Study ◽  
Cetylpyridinium Chloride ◽  
Ruthenium Red ◽  
Heart Tube ◽  
Fibrillar Material ◽  
Low Concentrations ◽  
Dorsal Mesocardium ◽  
Myocardial Layer

The hearts of chick embryos of stages 9–13 were subjected to SEM, TEM and histochemical studies to ascertain possible regional differences in the structure and composition of the cardiac jelly. Two distinct regions, the cardiac jelly filling the space located between the myocardium and the endocardium (MECJ) and the cardiac jelly filling the dorsal mesocardium (EECJ), were distinguished by their structural and histochemical properties. MECJ is formed by amorphous and fibrillar material arranged between the endocardial and myocardial layer. The amount of its components increases when cetylpyridinium chlorideis introduced into the fixative, and it appears intensely stained by ruthenium red and alcian blue at low concentrations of MgCl2. The amount and arrangement of its componentsincrease during the beginning of the looping process of the heart tube. The EECJ is very rich in ruthenium-red-positive basal-lamina-like material and the addition of cetylpyridinium chloride to the fixative does not modify its appearance. It also appears poorly stained by alcian blue at low concentrations of MgCl2 and its arrangement undergoes modifications closely associated with the events of endocardial fusion. The possible significance of these results in the early morphogenesis of the heart is discussed.

Cell Surface Glycoconjugates of Euglena gracilis (Euglenozoa): Modifications under Potassium and Magnesium Deficiency

1997 ◽  
Vol 52 (1-2) ◽  
pp. 33-41
Author(s):  
Angelika Preisfeld ◽  
Gabriele Scholten-Beck ◽  
Hans Georg Ruppel
Keyword(s):  
Cell Surface ◽  
Protein Content ◽  
Alcian Blue ◽  
Euglena Gracilis ◽  
Magnesium Deficiency ◽  
Ruthenium Red ◽  
Complete Medium ◽  
Cell Surface Changes ◽  
Cell Surface Glycoconjugates ◽  
Surface Changes

Abstract Biochemical and ultrastructural examinations on the pellicle of autotrophically grown Eu­glena gracilis were carried out after three days under potassium and magnesium deficiency. Cell-surface changes were detected by lectin assay. Compared to cells grown in complete medium, deficient cells become larger in shape, accompanied by rising carbohydrate, chloro­phyll and protein content, bind more and other lectin molecules: an increase of mainly galactose and N-acetylgalactosamine receptors was observed. Investigations with the mucilage stains alcian blue and ruthenium red indicated that mucilaginous material is released under deficient conditions, whereas the control cells show a strong precipitate of these stains well inside the cells beneath the pellicle.

scholarly journals Abnormal mast cell granules in the beige (Chédiak-Higashi syndrome) mouse.

1975 ◽  
Vol 23 (2) ◽  
pp. 117-122 ◽  
Author(s):  
E Y Chi ◽  
D Lagunoff
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Alcian Blue ◽  
Toluidine Blue ◽  
Ruthenium Red ◽  
Chediak Higashi Syndrome

Mast cells of beige (C57BL/6J) (bg-j/bg-j) mice were examined histochemically and ultrastructurally. Mast cell granules in the beige mice were markedly enlarged and irregular in shape. Granule contents stained uniformly with acidified toluidine blue, but with ruthenium red and Alcian Blue-safranin, two components were evident. The rims of the abnormal granules stained with ruthenium red and with Alcian Blue; the centers of the granules were clear with ruthenium red and stained with safranin. Mast cell granules thus represent another abnormal organelle in the Chédiak-Higashi syndrome.

scholarly journals Electron histochemistry and ultrastructural localization of carbohydrate-containing substances in the sheath of Volvox.

1976 ◽  
Vol 24 (5) ◽  
pp. 668-673 ◽  
Author(s):  
M D McCracken ◽  
W J Barcellona
Keyword(s):  
Periodic Acid ◽  
Ultrastructural Localization ◽  
Ruthenium Red ◽  
Cationic Dyes ◽  
External Layer ◽  
Colloidal Iron ◽  
Electron Histochemistry ◽  
Reactive Groups ◽  
Sheath Material

The location and characteristics of carbohydrate-containing structures within the intact sheath of Volvox were studied by 3,3'-diaminobenzidine tetrahydrochloride-osmium, colloidal iron, colloidal thorium, ruthenium red and periodic acid-silver methenamine staining. The sheath consists of external and internal fibrillar layers separated by a tripartite structure. The external layer reacts positively with 3,3'-diaminobenzidine tetrahydrochloride, colloidal iron, colloidal thorium and ruthenium red, indicating that it contains acid mucosaccharides. Staining in the external layer is abolished by Ba(OH)2 treatment. The tripartite structure and internal fibrillar layer contain periodic acid reactive groups which do not occur in the external layer. Under certain conditions, reactions between the cationic dyes and the internal material were also observed. It is postulated that the internal matrix of the sheath contains glycoproteins or a mixture of acid mucosaccharides and glycoproteins. Possible functions of the sheath material are discussed.

New methods to simultaneously localize proteoglycan by LM and EM

1978 ◽  
Vol 36 (2) ◽  
pp. 672-673
Author(s):  
Nona Shepard
Keyword(s):  
Articular Cartilage ◽  
Electron Micrographs ◽  
Toluidine Blue ◽  
Chemical Interaction ◽  
Ruthenium Red ◽  
Cationic Dyes ◽  
Keratan Sulphate ◽  
New Methods ◽  
Pathological Conditions ◽  
Sectioned Tissue

Proteoglycan is a major component ofa cartilage and up to 70% can be lost during routine processing unless special measures are taken. With this in mind and the fact that proteoglycan loss and or alteration is primary in most pathological conditions of articular cartilage, full retention is, therefore, essential to validate the interpreting of electron micrographs.The cationic dyes, Safranin 0 and Toluidine Blue 0, seemed like likely candidates as their chemical interaction with proteoglycan has been well documented by Rosenberg. Both dyes bind stoichlometrically to polyanions; one molecule of dye to each negatively charged chondroitin 6 - sulfate or keratan sulphate molecule. Szirmal has reported that cationic dyes bind proteoglycan of fresh sectioned tissue. Therefore, following Insitu fixation with Safaranin 0 or Toluidinz Blue 0, thz resulting Image would be a quantitative example of proteoglycan localization and if present in sufficent mass would scatter electrons. Ruthenium red is widely used for E. M., but we found it of little value as an L. M. stain

scholarly journals Preservation of cartilage matrix proteoglycans using cationic dyes chemically related to ruthenium hexaammine trichloride.

1992 ◽  
Vol 40 (7) ◽  
pp. 909-917 ◽  
Author(s):  
E B Hunziker ◽  
A Ludi ◽  
W Herrmann
Keyword(s):  
Osmium Tetroxide ◽  
Ruthenium Red ◽  
Cartilage Matrix ◽  
Cationic Dyes ◽  
Cartilage Tissue ◽  
Tissue Sections ◽  
Aldehyde Fixation ◽  
Violet Color ◽  
Black Color

We tested various cationic dyes chemically related to ruthenium hexaammine trichloride (RHT) [i.e., the RHT-cyclohexanedione complex (RHT-CC), pentaamine ruthenium N-dimethylphenylenediimine trichloride (PRT), tris-(bipyridyl)ruthenium (II) chloride (TRC), tris (bipyridyl) iron (II) chloride (TIC), and cobalt hexaammine trichloride (CHT)] for their effectiveness in precipitating cartilage matrix proteoglycans in situ. Dyes were introduced into media at the onset of processing and were present throughout both aldehyde fixation and osmium tetroxide post-fixation. Contrary to expectation, most of the dye-proteoglycan complexes generated and stable under aldehyde fixation conditions were found to be unstable during post-fixation despite the continuing presence of the dye. A similar phenomenon was also found for the cationic dyes commonly used for precipitation of proteoglycans in cartilage tissue sections (such as Acridine Orange, Alcian Blue, Azure A, Methylene Blue, and Ruthenium Red). Only two dyes, i.e., RHT and the newly tested RHT-CC, formed proteoglycan precipitates sufficiently stable to resist disruption and extraction during osmium tetroxide post-fixation. The latter may be particularly useful in semiquantitative analyses of proteoglycan content in unstained tissue sections owing to its intense brown-black color. For applications in which the osmium tetroxide post-fixation step may be omitted, TRC and PRT may also be valuable for semiquantitative histochemistry by virtue of their stable fluorescence and intense violet color signals, respectively.

Sign in / Sign up

Export Citation Format

Share Document