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At least 19 recordsLinked to original sources
PubMed1977

The Alcian blue and combined Alcian blue--Safranin O staining of glycosaminoglycans studied in a model system and in mast cells.

Polyacrylamide films containing different glycosaminoglycans have been applied to the study of the Alcian Blue and combined Alcian Blue--Safranin O staining procedures. It was found that the polyacrylamide matrix can be interpreted as some kind of 'barrier' around the substrate molecules, a situation which can be compared to a certain extent with what occurs in situ, where complex protein molecules can likewise form a 'barrier'. The Alcian Blue staining of the model films was found to follow the Lambert-Beer law. The time to reach optimal dye binding depended on the concentration of the glycosaminoglycan enclosed in the model films and on the concentration of Alcian Blue in the dye solution. Lowering the pH of the dye solution appeared to increase the rate of staining. Optimal staining of model films in the presence of salt or urea was not possible, because under these conditions the pores of the polyacrylamide matrix became blocked. Alcian Blue was found to bind irreversibly to the glycosaminoglycan molecules enclosed in the polyacrylamide films. The results of the combined Alcian Blue-Safranin O staining applied to model films appeared to be highly dependent on the amount of Alcian Blue bound to the glycosaminoglycan in the first step of the double staining procedure. No specific differences were noticed between the behaviour of the different glycosaminoglycan-Alcian Blue complexes towards the Safranin O binding in the next step. As the theoretical basis for the application of the combined Alcian Blue-Safranin O staining was also found not to be completely valid, the conclusion was reached that this double staining cannot be used for the histochemical identification of glycosaminoglycans. The colour retained by a certain glycosaminoglycan-containing part of the specimen only delivers information about the accesibility of that part for Alcian Blue.

Alcian Blue↗
PubMed1987

Alcian blue-alcian yellow mapping of neurosecretory cells in the central nervous system of the salt marsh pulmonate snail Melampus bidentatus.

1. The neurosecretory system of the primitive ellobiid Melampus bidentatus (Pulmonata: Basommatophora) was investigated using Alcian blue-Alcian yellow histochemistry. 2. Putative neurosecretory cells within the central ganglia were distinguished by the criteria of cell size, position, and staining reaction. 3. The cerebral ganglia, with attached lateral lobes, contained the greatest diversity of neurosecretory cell types (at least seven), including single cells and cell clusters ranging from two to 40 cells. 4. Four neurosecretory cell types were identified in the parietal and visceral ganglia, two in the pedal ganglia, and one each in the buccal and pleural ganglia. 5. Neurosecretory system homology among pulmonate gastropods is suggested by the close similarity of the Melampus AB/AY cell map to those reported in the literature for two limnic basommatophorans and for four terrestrial stylommatophorans.

Alcian Blue↗
PubMed1980

A comparative study on neurosecretion demonstrated by the alcian blue-alcian yellow technique in three terrestrial pulmonates (Stylommatophora).

Neurosecretion was studied in the central ganglia of three terrestrial pulmonates (Deroceras reticulatum, Arion hortensis, and Helix aspersa) with the alcian blue-alcian yellow (AB/AY) staining method. A number of neurosecretory cell types were distinguished in all three species (D. reticulatum: 9, A. hortensis: 11, H. aspersa: 13) With AB/AY the cells stain different shades of green and yellow. The histochemical differences reflect ultrastructural differences: the cell types contain elementary granules of different size and appearance. Some cell types form distinct groups of up to 50 cells, while others occur in smaller groups or as single cells. Neurosecretory axons could only be traced in a few cases to the probable neurohaemal areas. Based on the following criteria: (1) staining properties of cells (2) size of cells, (3) position of cells in the ganglia, it appears that the neurosecretory systems of the three species show great similarity, although some differences are apparent.

Animals↗
PubMed1994

Mucopolysaccharidoses screening: dimethylmethylene blue versus alcian blue.

The dimethylmethylene blue (DMB)-based screening procedure for mucopolysaccharidosis and the alcian blue (AB)-based procedure both measure glycosaminoglycan (GAG) content directly in urine. We compared the two procedures. Absorbance per microgram of GAG for DMB was 25 times that obtained with AB, resulting in a requirement for 10 times smaller sample volumes for the former. Recoveries of added heparan sulphate for the AB assay in the absence and presence of added protein (2.5 g/L) were comparable (78-95% and 75-111%), as was the case with the DMB assay. Here, recoveries were generally better (94-103% and 96-100%). The coefficient of correlation (CC) between the two methods measured on 86 reference urine samples was 0.861. For mucopolysaccharidosis urines the CC was higher (0.928). Age-dependent reference values were comparable. Performance of the assays was compared by measurement of GAG content in 24 mucopolysaccharidosis urines. Standard deviation scores (measured value minus mean of reference values divided by standard deviation in reference values) and probabilities for classification in the patient group were calculated and compared. Especially for urines from patients with less distinctly increased GAG content (Sanfilippo, Scheie), scores and probabilities were higher with the DMB assay.

Adolescent↗
PubMed1978

A permanent cell viability assay using alcian blue.

The alcian blue dye exclusion method for glutaraldehyde-fixed cells has been utilized with "centrifugal cytology" to prepare permanent records of the viability of individual cells present in suspensions. The viability of spleen cell suspensions separated by linear bovine serum albumin density gradient centrifugation has been measured with this method. Combined light and scanning electron microscopy of nonviable and viable cells demonstrated membrane alterations in alcian blue-stained nonviable cells, while viable cells were spherical and displayed uniform surface features.

Alcian Blue↗
PubMed1993

Simultaneous preparation and quantitation of proteoglycans by precipitation with alcian blue.

Conditions for specific interaction between Alcian blue and proteoglycans were optimized by comparing the differential spectra of Alcian blue obtained with purified chondroitin sulfate dissolved in water with the spectra obtained with nasal cartilage proteoglycans dissolved in synovial fluid. A method was then designed that provides specific precipitation of proteoglycans or glycosaminoglycans in 4 M guanidine - HCl in the presence of protein, hyaluronic acid, or nucleic acids. The specificity is achieved by using a low pH in combination with detergent and high salt concentration. Stepwise addition of reagents is necessary to avoid binding of Alcian blue to proteins and nucleic acids. All polyanions, except polysulfates, are first neutralized by lowering the pH to 1.5. By including detergent in this step, the hydrophobic protein regions are blocked and not accessible for binding with the dye. These regions could otherwise bind Alcian blue by hydrophobic interaction. When the Alcian blue reagent is added after, only the polysulfated molecules will remain charged and free to interact with Alcian blue. At least 0.4 M guanidine-HCl is required to abolish the negative interference by proteins. All sulfated glycosaminoglycans are precipitated at 0.4 M guanidine-HCl. With increasing guanidine-HCl concentrations, the different glycosaminoglycans are precipitated in accordance with the critical electrolyte concentration of the respective glycosaminoglycan. The Alcian blue precipitation can be performed at different concentrations of guanidine-HCl in order to separate different classes of proteoglycans. Excess dye and contaminating proteins are removed by a wash in a DMSO-MgCl2 solution and the precipitate is dissolved in a mixture of guanidine-HCl and propanol. For quantitation, the absorbance is recorded in a microplate reader with the 600-nm filter, the assay being linear between 0.5 and 20 micrograms proteoglycan. Since no digestion of samples with protease is needed, the proteoglycans are recovered in native form. The proteoglycan-Alcian blue complexes dissociate in the guanidine-HCl/propanol mixture and the proteoglycans can be selectively precipitated with propanol. The dye is used for quantitation and the proteoglycans can be utilized for further analysis.

Alcian Blue↗
PubMed1975

Staining of sulphatides in metachromatic leukodystrophy with Alcian blue at high salt concentrations.

Alcian blue combines with purified sulphatide in 1.OM magnesium chloride. In tissue sections from patients with metachromatic leukodystrophy, sulphatide is stained by Alcian blue in 0.8 M magnesium chloride, and the staining can be abolished by prior treatment with chloroform and methanol. The simplicity of the technique, its specificity and ease of interpretation recommend Alcian blue staining at high salt concentrations as a routine method in the diagnosis of metachromatic leukodystrophy.

Alcian Blue↗
PubMed1982

Monastral fast blue. Cytochemical properties of a reaction product from Alcian blue stained chromatin.

The staining of chicken erythrocyte nuclei with Alcian blue followed by a treatment with ethanol-NaOH, showed to be highly resistant to acid extraction (5 N HCl) due to the alkaline conversion of Alcian blue into the insoluble pigment Monastral fast blue. It is assumed that the intercalative binding of the planar Monastral fast blue molecule to DNA accounts for its competition effect toward the intercalating fluorochromes acridine organe, ethidium bromine and daunomycin.

Alcian Blue↗