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At least 19 recordsLinked to original sources
PubMed1977

Age changes in Alcian blue staining of glycosaminoglycans in sheep articular cartilage.

Alcian blue staining of glycosaminoglycans in sheep articular cartilage is described and discussed. Chodrocytes contain mainly low molecular weight chondroitin sulphate in the outer layers but keratan sulphate increases with depth. Protein-polysaccharide interaction increases with age but the cellular staining decreases in intensity with age. Lacunar capsules contain material of higher molecular weight, much of which is chondroitin sulphate, but no protein-polysaccharide interaction occurs. The matrix of the outer layers is deeply stained while that of the inner layer is pale and consists mainly of keratan sulphate. When stained in high salt concentrations (greater than 0-7 M) the matrix of the middle layer shows two zones; hyaluronidase treatment allows these zones to be differentiated at lower salt concentrations.

Aging↗
PubMed1977

Enzyme clearing of alcian blue stained whole small vertebrates for demonstration of cartilage.

Preparation of small vertebrates cleared after alcian blue staining of cartilage is facilitated by trypsin digestion. Specimens are fixed in formation, washed, skinned, and eviscerated. After staining in a solution of alcian blue in acetic acid-alcohol for 24-48 hours, they are transferred to water through graded alcohols. Excess alcian blue is removed over a period of up to three weeks by changes every 2-3 days of 1% trypsin in approximately one-third-saturated sodium borate. Bony tissues may be stained after this in a solution of alizarin red S in 0.5% KOH. Specimens are bleached if necessary and dehydrated through graded KOH-glycerine mixtures for storage in glycerine. Since alcohol treatment in addition to formalin fixation does not affect results with this method, it should be useful to researchers who want to study the cartilage or cartilaginous skeletons in museum specimens, which are routinely fixed in formalin and stored in alcohol.

Alcian Blue↗
PubMed1975

Alcian Blue staining of cartilage for electron microscopy. Application of the critical electrolyte concentation principle.

The critical electrolyte concentration principle was applied to the Alcian Blue staining of rat epiphyseal cartilage proteoglycans for electron microscopy. The distribution and structure of material in glutaraldehyde-fixed cartilage stained at pH 5.8 without MgCl2 and in the presence of 0.05, 0.4, 0.5, 0.9 and 1.0 M MgCl2 was compared with that produced by simultaneous staining and fixation at neutral pH. Both methods resulted in staining of intracellular material within vacuoles as well as staining of non-collagenous matrix material. The structure and distribution of Alcian Blue-positive matrix material consisted of rounded or polygonal granules which accumulated around cells in the proliferative and hypertrophied zones. A similar pattern of distribution was observed in samples stained in the presence of 0.4 or 0.5 M MgCl2. In these cases, however, the stained material exhibited a ribbon-like configuration and granules were few in number. Increasing the MgCl2 concentration to 1.0 M resulted in a marked reduction of Alcian Blue stained material. No ribbon-like structures were observed, and matrix granules were reduced in both number and size. The decreased staining associated with increased electrolyte concentration lends support to the concept that epiphyseal cartilage matrix granules are composed primarily of chondroitin sulphate, and suggest that this same material is present in vacuoles associated with the Golgi apparatus in chondrocytes of the proliferative and hypertrophying zones.

Alcian Blue↗
PubMed1978

Alcian blue staining intestinal goblet cell antigen (GOA): a marker for gastric signet ring cell and colonic colloidal carcinoma.

An Alcian blue staining, perchloric acid-soluble, antigenic acidic mucosubstance (GOA) was purified from human gastric signet ring cell carcinoma with DEAE-cellulose chromatography, Seqhadex G-200 and preparative polycrylamide gel electrophoresis. Specific antisera were raised which reacted in indirect immunoenzyme histology with normal goblet cells of the small and large intestine and with goblet cells of intestinalized gastric mucosa. In surgical resection specimens of the stomach (n = 100) and of the colon (n = 19) 3 gastric signet ring cell carcinomas and 3 colonic colloidal adenocarcinomas stained for GOA, demonstrating an immunochemical relationship with normal intestinal goblet cells.

Adenocarcinoma, Mucinous↗
PubMed1976

Experience with the sodium sulphate-Alcian Blue stain for amyloid in cardiac pathology.

The sodium sulphate-Alcian Blue (SAB) method, which stains amyloid green, was evaluated in 220 hearts from elderly patients. The technique proved superior to the Congo red, crystal violet, and thioflavine T methods used either singly or as a battery for the demonstration of cardiac amyloid. Amyloid was easily identified under the X3 objective, even in small amounts. A few non-amyloid components stained varying shades of green but were easily distinguished on morphological grounds. No false positive or equivocal reactions occurred, and in particular elastic laminae and paravascular connective tissue were not tinctorially confused with amyloid. The SAB stain is technically simple and consistently reproducible, and no special light source is required for examination. An additional advantage in cardiac pathology is the simultaneous demonstration of any fibrosis, basophilic myofibre degeneration, tissue mast cells and mucoid degeneration of valves present.

Aged↗
PubMed1975

A comparison of aldehyde fuchsin and alcian blue staining of neurosecretory material in Oncopeltus fasciatus.

The dynamics of the "A" cells of the parsintercerebralis of Oncopeltus fasciatus over the first eight days of adult life was studied by microspectrophotometry of sections stained either with aldehyde fuchsin or alcian blue 8 GX. The data show that the two stains differ in their selectivity as they record different events in the history of the cells. A hypothesis is proposed that the aldehyde fuchsin is more sensitive to the presence of a "carrier" protein in the cell, whereas alcian blue 8 GX is more sensitive to the presence of the "active principle" in the cell.

Alcian Blue↗
PubMed1977

Distribution of mucosubstances in adenoid cystic carcinoma.

The distribution of mucosubstances in adenoid cystic carcinoma was investigated, and an attempt was made to characterize histochemically the various mucosubstances present. For these purposes the high iron diamine technique (HID), as well as the Astra blue, aldehyde fuchsin and Alcian blue staining methods were employed. Alcian blue was further combined with the periodic acid-Schiff (PAS) technique, the Alcian blue being applied at pH levels between 0.5 and 2.5. In addition the effect of neuraminidase and hyaluronidase treatment as well as methylation and acid hydrolysis procedures on the staining qualities were studied. Acidic mucosubstances with varying histochemical properties were present in different structures of the neoplasm. The characteristic pseudocyst, a major structural component of the neoplasm, stained strongly with HID, Astra blue, aldehyde fuchsin and Alcian blue at low pH. These staining reactions were markedly suppressed by hyaluronidase treatment, and are apparently attributable to the presence of chondroitin 4- and/or 6-sulfate. Employing the Alcian blue-critical electrolyte concentration technique, the basophilia of the pseudocysts was suppressed at a concentration of 0.5-0.6 M MgCl2, which might indicate polysaccharides of relatively low degree of sulfation. An additional, non-sulfated acid mucin could also be demonstrated in these structures. In certain duct and gland like structures of the tumours, a change in staining pattern from blue or blue-red to red could be observed after exposure of the sections to neuraminidase and subsequent staining with the Alcian blue (pH 2.5)-PAS sequence. Similar observations were also made when the pH of the Alcian blue was lowered to 1.5-1.0, as well as after acid hydrolysis. These findings afford evidence for the presence of a neuraminidase susceptive sialomucin in certain epithelial secretions of the tumor. At the ultrastructural level the replicated basement lamina of the pseudocysts displayed a strong positive reaction with the PA-CrA-silver staining technique. Furthermore, amorphous material within the lumina of small duct like structures also displayed a positive reaction. The amorphous material of the cystic compartments was less reactive.

Carcinoma, Adenoid Cystic↗
PubMed1978

Regional differences in mesenchymal cell morphology and glycosaminoglycans in early neural-fold stage rat embryos.

Rat embryos with two to four pairs of somites (day 9 of gestation) were examined by scanning electron microscopy and Alcian blue staining. The neural folds, which represent only future brain region at this stage, form a pair of elongated hemispheres with a deep neural groove between them. In transverse section the neural ectoderm is biconvex; the cranial mesenchyme cells beneath them are widely separated by extracellular matrix (ECM) and are joined to each other and to the ectodermal basement membrane by fine cytoplasmic processes and strands of ECM material. In contrast, mesenchyme cells close to the primitive streak are closely packed, having broad areas of surface contact and only small amounts of ECM. The nature and distribution of ECM, cell surface, and basement membrane glycosaminoglycans (GAG) were investigated by staining with Alcian blue at specific pH values in combination with enzyme pretreatments, and at various critical electrolyte concentrations. The results indicate that the GAG of the ectodermal basement membrane, mesenchymal ECM and mesenchymal cell surfaces are in continuity with each other and consist largely of hyaluronate and chondroitin sulphates. Differences in morphology and histochemistry of neural fold and primitive streak regions are discussed in relation to their possible morphogenetic significance.

Animals↗
PubMed1975

The diagnosis of chronic simple conjunctivitis. Vital staining of tarsus with tetrazolium - alcian blue mixture.

Vital staining with a tetrazolium - alcian blue mixture gave a characteristic punctate red staining of the superior or the inferior tarsus in 69% of all examined patients with chronic simple conjuctivitis (69 patients subjected to 168 examinations). The superior tarsus was more frequently stained than the inferior (61% against 33%). The staining was most often concentrated centrally or in the middle anterior part of the superior tarsus. If located on the inferior tarsus, it was most often found medially, anteriorly. No corresponding staining was seen in normal eyes. The differential diagnosis involving keratoconjunctivitis sicca and pemphigoid is mentioned. The staining is due to enzymatic reduction of tetrazolium in the cytoplasm of the tarsal epithelial cells. The positive vital staining finding is consistent with a diagnosis of chronic simple conjunctivitis.

Alcian Blue↗
PubMed2025

Impact of stromal maturity and proportion on prognosis and immune landscape in colorectal cancer.

BACKGROUND: Tumour microenvironment and cancer cells have constant interaction affecting cancer progression. Tumour-stroma ratio (TSR) in the tumour centre and desmoplastic reaction (DR) classification at the invasive margin are prognostic factors based on stroma evaluation on H&E slides. However, their combined value and immunological associations remain poorly defined. This study examines the prognostic and immunological value of TSR, DR, and their combination in two large colorectal cancer cohorts. METHODS: Two colorectal cancer cohorts (N&#x2009;=&#x2009;1,876) were analyzed. We introduced a three-tiered Stromal Maturity and Proportion Score (SMAPS) based on the presence of high (>50%) TSR and myxoid stroma (immature DR classification). Alcian blue staining was used to further quantify myxoid stroma. Multiplex immunohistochemistry combined with digital image analyses, was utilized to study immune cell densities associated with SMAPS, TSR, DR, and Alcian blue intensity. RESULTS: In the study cohort (N&#x2009;=&#x2009;1,100), SMAPS was a stronger predictor of cancer-specific mortality [HR for high (vs. low) SMAPS 2.01 (95% CI 1.47-2.75), p&#x2009;<&#x2009;0.0001] compared to TSR [HR for stroma-high (vs. stroma-low) 1.49 (95% CI 1.15-1.93), p&#x2009;=&#x2009;0.003] and DR classification [HR for immature (vs. mature) 1.84 (95% CI 1.39-2.45), p&#x2009;<&#x2009;0.0001]. High SMAPS, stroma-high TSR, and immature DR correlated with lower densities of CD3+ T cells, B cells, M1-like macrophages, CD66B+ granulocytes, and mast cells. Alcian blue staining was associated with immature DR and corresponding immune cells. The validation cohort (N&#x2009;=&#x2009;776) confirmed the association of SMAPS with survival and T cell densities. CONCLUSIONS: TSR and DR are independent prognostic factors for cancer-specific survival. SMAPS is a promising prognostic tool that integrates stromal maturity at the invasive margin and stromal proportion in the tumour centre. SMAPS has stronger prognostic value compared to TSR and DR classifications alone. A high stromal proportion and myxoid content are associated with an immunosuppressive microenvironment characterized by lower densities of antitumourigenic immune cells.

Humans↗
PubMed1975

Basophil counting with a new staining method using alcian blue.

Difficulties in obtaining reproducible and accurate enumeration of circulating basophils with existing techniques have hampered investigation of this infrequent cell population. A new basophil staining method is described that employs alcian blue dye for staining of heparin within basophils at low pH and in the presence of lanthanum ions. Basophil recognition is facilitated by reducing nonspecific nuclear staining. This objective is achieved because of the differences in stability of alcian blue-heparin, alcian blue-nucleic acid, lanthanum-heparin, and lanthanum-mucliec acid complexes. Reduction of pH after staining also favors solubilization of leukocyte cytoplasmic proteins, providing greater contrast between stained and unstained cells by reducing light scattering of the unstained leukocytes . The alcian blue staining method is suitable both for chamber basophil counting and automated basophil counting using continuous-flow sampling and electro-optical detection. The new staining method was evaluated by comparing it with the chamber counting method using toluidine blue in a triple-blind study in which the results of basophil counting by the alcian blue chamber method, alcian blue automated instrument method, and the toluidine blue chamber method were analyzed for reproducibility and compared with an indirect basophil count obtained from a 1000-cell leukocyte differential and a total leukocyte count. Both alcian blue staining methods gave greater reporducibility that toluidine blue and were more accurate, as evidinced by a significantly higher correlation with the indirect basophil count. The improved reproducibility, accuracy, and convenience of this method over existing methods should facilitate the collection of more meaningful information about circulating basophil levels in health and disease.

Alcian Blue↗
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↗
PubMed1978

Sexual differences among duodenal glands of hamster.

This histochemical study was undertaken to determine whether sexual differences existed in the duodenal glands of hamster. Alcian blue staining and atomic absorption spectrophotometry (for copper content of the bound Alcian blue) showed the duodenal glands of males to contain about double the acid mucosubstance of the female glands. Periodic acid Schiff staining was stronger in the females over a larger portion of the gland than in males. It was concluded that sexual differences were evident in duodenal glands of hamster.

Animals↗
PubMed1975

The effect of short-term immobilization on the rabbit knee joint cartilage. A histochemical study.

Rabbit knee joints were immobilized in the extended position for 1 to 6 days, and examined by means of histochemical techniques. In the articular cartilage matrix, the earliest change was the loss of affinity for metachromatic stains, confirming earlier observations. The affinity of cartilage for high iron diamine and alcian blue stains was also diminished, although less dramatically; these stains demonstrated the concentration of glycosaminoglycans more accurately than metachromatic stains. With the colloidal iron method, changes were seen in the chondrocytes before there was any loss of affinity for metachromatic stains. Following only 1 or 2 days of immobilization, the chondrocytes in the gliding surface of the joint stained intensely with colloidal iron whereas those in the controls stained very minimally.

Alcian Blue↗
PubMed1978

Responses of lysosomes in the digestive cells of the common mussel, Mytilus edulis, to sex steroids and cortisol.

Estradiol-17beta and progesterone at physiological concentrations in vivo induced a reduction in lysosomal stability in the digestive cells of Mytilus edulis. Estradiol-17beta (10(-8) M) also reduced lysosomal stability within 15 min in vitro. Lysosomal stability was determined cytochemically as the labilisation period for latent N-acetyl-beta-hexosaminidase and this was shown to be inversely related to microdensitometric measurements of staining intensity for this enzyme. Estradiol-17beta did not appear to induce complete labilisation or cytochemical activation of lysosomal hexosaminidase and a second, much longer labilisation period could be determined for this hormone. The effects of estradiol-17beta were partially counteracted by cortisol (10(-2) M). There was an increase in PAS staining of secondary lysosomes and an increase in alcian blue staining of residual bodies in digestive cells of animals exposed to estradiol-17beta, while no changes could be observed in basophil cells. The significance of these results is discussed in terms of the physiological role of digestive cells and their possible function as target cells for estradiol-17beta and progesterone.

Animals↗
PubMed1978

Topographical distribution of sulfated glycosaminoglycans in the surface layers of the human temporomandibular joint. A histochemical study of an autopsy material.

The right temporomandibular joint (TMJ) was removed at autopsy from 18 individuals and the articular surfaces were scored for macroscopic lesions. Soft tissue specimens were cut out of the medial, lateral and posterior parts of the temporal and condylar components and frozen. The sections were stained with toluidine blue (1-N HCl solution) and with alcian blue (CEC method) for sulfated glycosaminoglycans. The sections were examined for inter- and pericellular metachromasia and alcian blue staining. Macroscopic surface lesions were found in 14 of the joints. The highest score of interand pericellular metachromasia was found in the anterior parts of the joints and a negative correlation was found between the score of surface lesions and that of intercellular metachromasia. Pericellular metachromasia was found mainly in the mineralized cartilage and cartilage layers of the articular surface. Although the CEC values of pericellular staining commonly reached a level indicating the presence of keratan sulfate, the majority of the CEC values found corresponded to those of chondroitin-dermatan sulfate. It was concluded that sulfated GAG are localized mainly to the anterior part of the TMJ and that macroscopic TMJ surface lesions are associated with a reduction of sulfated glycosaminoglycans.

Adolescent↗
PubMedDate not supplied

Morphological changes of tracheobranchial mucosa in dogs caused by breathing through a surgically produced tracheal fistula.

The author studied the morphological changes developing in the tracheobronchial mucosa following a surgical tracheal fistula. The experiments were carried out on 19 dogs which were divided into 2 groups. Group I included 10 dogs, in which specimens were obtained by bronchoscopy; in group II, 9 dogs, specimens were obtained directly. Histological examinations were carried out by Gomori's reaction and hematoxylin-eosin staining method. Histo-chemical tests included PAS reaction and alcian blue stain. The results showed that after 24 hours morphological changes mainly involved the tracheal mucosa and their characteristic feature was the variety of forms of lesions of the epithelium covering the mucosa. The longer the duration of the fistula the more these changes increased in intensity and involved further parts of the tracheobronchial tree. The author also took into account the living conditions in the kennels where the dogs were kept following the tracheostomy. The changes observed were compared with those reported by other authors and the correlation between the occurrence of these changes and the environmental conditions and length of breathing through the surgically produced fistula was analysed.

Animals↗