Dual staining of some sulfated mucopolysaccharides with alcian blue (pH 1.0) and ruthenium red (pH 2.5)

1970 ◽  
Vol 23 (1) ◽  
pp. 13-20 ◽  
Author(s):  
Kazuyori Yamada
Keyword(s):  
Alcian Blue ◽  
Ruthenium Red ◽  
Dual Staining

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 DUAL STAINING OF MAST CELL CYTOPLASMIC CONSTITUENTS BY ALCIAN BLUE AND SAFRANIN

1962 ◽  
Vol 10 (4) ◽  
pp. 503-503 ◽  
Author(s):  
W. CURTIS WORTHINGTON ◽  
NAN C. BAILEY
Keyword(s):  
Mast Cell ◽  
Alcian Blue ◽  
Dual Staining

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.

Concurrent validity of biofilm detection by wound blotting on hard-to-heal wounds

2021 ◽  
Vol 30 (Sup4) ◽  
pp. S4-S13
Author(s):  
Adam Astrada ◽  
Gojiro Nakagami ◽  
Takeo Minematsu ◽  
Taichi Goto ◽  
Aya Kitamura ◽  
...  
Keyword(s):  
Alcian Blue ◽  
Concurrent Validity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ruthenium Red ◽  
Extracellular Dna ◽  
Wound Management ◽  
Blue Staining ◽  
Clinical Samples

Objective: Wound biofilms delay healing of hard-to-heal wounds. Convenient biofilm identification tools for clinical settings are currently not available, hindering biofilm-based wound management. Wound blotting with biofilm staining is a potential tool for biofilm detection, owing to its convenience. Although predictive validity of wound blotting has been established, it is necessary to confirm its concurrent validity. Furthermore, current staining systems employing ruthenium red have some disadvantages for clinical use. This study aimed to evaluate the usability of alcian blue as a substitute for ruthenium red. Method: Both in vitro and in vivo clinical samples were used to investigate validity and usability. Results: The in vitro study showed that proteins and extracellular DNA in biofilms did not affect staining ability of ruthenium red and alcian blue in the detection of biofilms. In the in vivo study, using a wound biofilm model with Pseudomonas aeruginosa, the staining sensitivity of ruthenium red was 88.9% and 100% for alcian blue, with correlation coefficients of signal intensities with native polyacrylamide gel electrophoresis (PAGE) of r=0.67 (p=0.035) and r=0.67 (p=0.036) for ruthenium red and alcian blue, respectively. Results from clinical samples were r=0.75 (p=0.001) for ruthenium red and r=0.77 (p<0.001) for alcian blue. The sensitivities of wound blotting staining by ruthenium red and alcian blue were very high and had a good correlation with native PAGE analysis. Conclusion: Because the alcian blue procedure is more convenient than the ruthenium red procedure, wound blotting with alcian blue staining would be a promising tool to guide clinicians in delivering biofilm-based wound management.

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