scholarly journals DIAMINE METHODS FOR DIFFERENTIATING MUCOSUBSTANCES HISTOCHEMICALLY

1965 ◽  
Vol 13 (3) ◽  
pp. 211-234 ◽  
Author(s):  
S. S. SPICER
Keyword(s):  
Alcian Blue ◽  
Periodate Oxidation ◽  
Goblet Cells ◽  
Colloidal Iron ◽  
Mucous Layer ◽  
Para Isomer ◽  
High Iron ◽  
Aldehyde Fuchsin ◽  
Golgi Zone ◽  
Acid Mucosubstances

A mixture of N,N-dimethyl- m-phenylenediamine (HCI)2 with the para isomer at pH 3.5-4.0 stains acid mucosubstances selectively. Various sialomucins as well as sulfomucins differ in their reactivity with this mixed diamine reagent and in the manner in which the pH and ionic strength of the solution affect their staining. Prior periodate oxidation eliminates or markedly decreases the staining of some sulfo- and sialomucins but has no influence on the reactivity of others. Both sulfated mucosaccharides and sialomucins differ also according to whether or not their affinity for azure A, colloidal iron or alcian blue persists following periodate oxidation and exposure to the meta diamine. Azurophilia of nuclei at pH 4.0 is abolished by meta diamine used after Feulgen hydrolysis. Ferric chloride added to a solution of both diamines allows selective demonstration of most acid mucosubstances. This "low iron" diamine method followed by alcian blue stains sulfomucins and many sialomucins black, but other sialomucins blue. Some epithelia contain a mixture of blue and black-stained sialomucins. A variant technique uses a higher concentration of iron and diamines. This colors most sulfomucins black while leaving nonsulfated acid mucosubstances unstained. This high iron diamine method followed by alcian blue stains sulfomucins black and sialomucins blue. Results with the "high iron" diamine-alcian blue sequence indicate that sulfomucin occurs in some sites and sialomucin in others; in many epithelia a mixture of the two mucosubstances coexists either in the same on in different cells. Similar differentiation was obtainable also in many but not all sites with either the combination aldehyde fuchsin-alcian blue procedure on by assessment of differences between the results of alcian blue pH 2.5-PAS and alcian blue pH 1.0-PAS sequences. Applied to the surface mucous layer of certain intestinal and genitourinary epithelia, the methods differentiate areas of sulfomucin content alternating with others of sialomucin content. This is believed to indicate biogenesis of this secretion in the Golgi zone of nongoblet epithelial cells since staining of the surface mucous layer parallels that in the Golgi zone and often not that of neighboring goblet cells.

scholarly journals CYTOCHEMICAL LOCALIZATION OF ACID MUCOSUBSTANCES WITHIN RABBIT, CHICKEN, FROG AND FISH SKELETAL MUSCLE FIBERS

1969 ◽  
Vol 17 (1) ◽  
pp. 36-46 ◽  
Author(s):  
WILLIAM J. DOUGHERTY
Keyword(s):  
Alcian Blue ◽  
Skeletal Muscles ◽  
Phase Contrast ◽  
Blue Staining ◽  
Polarization Microscopy ◽  
Rabbit Muscle ◽  
Colloidal Iron ◽  
Cytochemical Localization ◽  
High Iron ◽  
Acid Mucosubstances

Skeletal muscles of rabbits, chickens, frogs and fishes were studied by bright field, phase contrast and polarization microscopy after treatment with: (1) Alcian Blue, pH 2.5, (2) Alcian Blue, pH 1.0, (3) low iron diamine, (4) high iron diamine, (5) dialyzed iron, pH 2.0, and (6) colloidal iron, pH 1.5. A prominent aspect of each of the muscles studied was transverse muscle bands which stained with Alcian Blue at pH 2.5, low iron diamine and dialyzed and colloidal iron reagents. Staining of transverse bands was not very intense with these reagents, suggesting that the materials demonstrated occurred in relatively low concentrations. Staining of transverse bands in chicken, frog and fish muscles was observed also after treatment with Alcian Blue at pH 1.0 or with high iron diamine. Staining of rabbit muscle with Alcian Blue at pH 1.0 or with high iron diamine was not detectable even after the most effective fixatives employed in this study, viz., Carony's fixative or buffered HgCl2. Phase contrast and polarization microscopy indicated that the transverse bands stained by the reagents employed corresponded to the I bands in agreement with previous electron cytochemical studies of skeletal muscles treated with dialyzed iron at controlled pH. Methylation for 4 hr at 60°C prevented I band staining with Alcian Blue and dialyzed iron in each of the muscles studied. Treatment of muscle sections with Vibrio cholerae neuraminidase, testicular hyaluronidase or ribonuclease under optimal conditions did not prevent Alcian Blue staining of I bands. Trypsin treatment of muscle sections did not prevent I band staining except when digestion was prolonged to the point that muscle striations were no longer recognizable. Saponification prior to hyaluronidase treatment reduced I band staining in each of the muscles studied. Saponification alone and saponification prior to neuraminidase digestion left I band alcianophilia intact. It was concluded that myofibrillar I bands of rabbit, chicken, frog and fish skeletal muscles contain acid mucosubstances. It was suggested that in each of these organisms an esterified, hyaluronic acid-like molecule is present within the I bands. Sulfated mucosaccharides occur as additional components in the I bands of fish, chicken and frog skeletal muscles; sulfated mucosaccharides apparently are not present in rabbit muscle I bands. Available evidence indicates that sialomucins are not present in the myofibrils of the organisms studied. The possible functional significance of I band acid mucosubstances in vertebrate skeletal muscles was discussed.

scholarly journals THE EFFECT OF DIGESTION WITH STREPTOMYCES HYALURONIDASE UPON CERTAIN HISTOCHEMICAL REACTIONS OF HYALURONIC ACID-CONTAINING TISSUES

1973 ◽  
Vol 21 (9) ◽  
pp. 794-803 ◽  
Author(s):  
KAZUYORI YAMADA
Keyword(s):  
Hyaluronic Acid ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Colloidal Iron ◽  
Periodic Acid Schiff ◽  
High Iron ◽  
Azure A ◽  
Aldehyde Fuchsin ◽  
Testicular Hyaluronidase

The effect of digestion with Streptomyces hyaluronidase upon certain histochemical reactions of hyaluronic acid-containing tissues has been studied in a series of human, mammalian and avian specimens. These histochemical reactions are those for the demonstration of neutral and sulfated and nonsulfated acid mucosaccharides such as periodic acid-Schiff, Alcian Blue (pH 1.0), azure A (pH 1.5), high iron diamine, aldehyde fuchsin, Alcian Blue (pH 2.5), Alcian Blue (pH 2.5)-periodic acid Schiff, azure A (pH 4.5), low iron diamine and colloidal iron. In addition, the effect of digestion with testicular hyaluronidase upon the same reactions of the same tissues was observed for comparison. Digestion with Streptomyces hyaluronidase diminishes the staining reactions due to hyaluronic acid but fails to affect those for neutral and sulfated acid mucosaccharides. These results indicate that digestion with Streptomyces hyaluronidase is a method of choice for the identification of hyaluronic acid in mucosaccharide histochemistry.

scholarly journals CORRELATED HISTOCHEMICAL STAINING AND S35O4= LABELING OF SALIVARY GLAND MUCOSUBSTANCES

1967 ◽  
Vol 15 (12) ◽  
pp. 745-751 ◽  
Author(s):  
T. JOHN LEPPI ◽  
SAMUEL S. SPICER ◽  
JACQUELINE G. HENSON ◽  
JOSEPHINE FIORAVANTI
Keyword(s):  
Salivary Gland ◽  
Connective Tissue ◽  
Salivary Glands ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Histochemical Staining ◽  
Periodic Acid Schiff ◽  
High Iron ◽  
Aldehyde Fuchsin

Correlated histochemical and autoradiographic studies were conducted on rat, rabbit, cat and dog salivary glands. Mucous acini of rat minor sublingual (SL) rabbit SL and cat posterior SL and submandibular (SM) glands were stained by Alcian Blue at pH 1.0, aldehyde fuchsin or the high iron diamine technique. The distribution of cells with this type of basophilia corresponded with that of cells shown autoradiographically to incorporate S35O4= and accordingly to contain sulfomucin. The lack of periodic acid-Schiff and alcohol-resistant metachromasia at pH 0.5 of rabbit SL mucin resembles connective tissue mucopolysaccharide. Some mucous acini of the dog posterior SL gland incorporate S35-sulfate and the distribution of isotope-labeled acini correlates with that of acini showing histochemical staining indicative of sulfomucin. The remainder of mucous acini in this gland do not show uptake of radiosulfate, but have alcianophilia that is reduced by sialidase digestion. All of the mucous acini of dog SM gland show correspondence of staining for sulfomucin with S35O4–, incorporation. The seromucous demilunes of this gland manifest sialidase-labile alcianophilia. The staining of sulfomucin and S35 labeling in each of the salivary glands studied are eliminated by a 4-hr methylation at 60°C.

High iron diamine?alcian blue mucin profiles in benign, premalignant and malignant colorectal disease

1988 ◽  
Vol 13 (4) ◽  
pp. 399-411 ◽  
Author(s):  
D. C. ALLEN ◽  
N. S. CONNOLLY ◽  
J. D. BIGGART
Keyword(s):  
Alcian Blue ◽  
Colorectal Disease ◽  
High Iron

scholarly journals Immunoreactivity of cytokeratins 7 and 20 in goblet cells and columnar blue cells in patients with endoscopic evidence of Barrett's esophagus

2009 ◽  
Vol 46 (2) ◽  
pp. 127-131 ◽  
Author(s):  
João Carlos Cantarelli Jr. ◽  
Renato Borges Fagundes ◽  
Luise Meurer ◽  
Marta Pires da Rocha ◽  
André Nicola ◽  
...  
Keyword(s):  
Intestinal Metaplasia ◽  
Barrett’S Esophagus ◽  
Alcian Blue ◽  
Barrett's Esophagus ◽  
Immunohistochemical Staining ◽  
Esophagogastric Junction ◽  
Cell Types ◽  
Goblet Cells ◽  
Cytokeratin 20 ◽  
Cytokeratin 7

CONTEXT: Barrett's esophagus is characterized by the presence of goblet cells. However, when alcian-blue is utilized, another type of cells, called columnar blue cells, is frequently present in the distal esophagus of patients with endoscopic evidence of Barrett's esophagus. Cytokeratin 7 and 20 immunoreactivity has been previously studied in areas of intestinal metaplasia at the esophagogastric junction. However, the expression of these cytokeratins in columnar blue cells has not been characterized. OBJECTIVE: To compare the expression of cytokeratin 7 and 20 in goblet cells and columnar blue cells in patients with endoscopic evidence of Barrett's esophagus. METHODS: Biopsies from 86 patients with endoscopic evidence of Barrett's esophagus were evaluated. The biopsies were stained for cytokeratin 7 and 20. RESULTS: Goblet cells were present in 75 cases and columnar blue cells in 50 cases. Overall, cytokeratin 7 expression was similar in goblet cells and columnar blue cells (P = 0.25), while cytokeratin 20 was more common in goblet cells (P <0.001). In individuals with both cell types, however, cytokeratin 7 staining was the same in goblet and columnar blue cells in 95% of the cases, and cytokeratin 20 staining was the same in 77%. CONCLUSION: Goblet cells and columnar blue cells have similar immunohistochemical staining patterns for cytokeratins 7 and 20 in patients with endoscopic evidence of Barrett's esophagus.

scholarly journals Mucus formation in goblet cells

1933 ◽  
Vol 113 (784) ◽  
pp. 459-463 ◽  
Keyword(s):  
Golgi Apparatus ◽  
Salivary Glands ◽  
Secretory Granules ◽  
Goblet Cells ◽  
Neutral Red ◽  
Basal Region ◽  
Mucous Cells ◽  
Cell Type ◽  
Golgi Network ◽  
Golgi Zone

The formation of mucus in goblet cells and its relation to the Golgi apparatus has been studied by various workers. Nassanow (1923) showed clearly that the mucin granules in the goblet cells of Triton originated in the Golgi apparatus, and so brought secretion in these cells into line with his theory of the bound secretion. More recently Clara (1926) has shown in the goblet cells of birds that the mucin first appears in the region next to the nucleus, between it and the gland lumen. Florey (1932, a, b ) has considered this more extensively in two recent papers, and for a number of mammals has shown that the mucin granules of goblet cells first form in the meshes of the Golgi network. In epithelial cells of the mouse vagina, undergoing conversion into mucous cells, he has found that the same process occurs. In a recent investigation of secretory formation in the salivary glands and pancreas it was found by the present author that in every cell type examined the young secretory granules first appeared in the basal region of the cell in relation to the mitochondria. Subsequent emigration occurred into the Golgi zone, where they underwent conversion into mature secretory granules. In the mucous cells of the salivary glands it was shown that these newly formed granules might be stained intravitam by Janus green or neutral red, and that in fixed preparations they stained selectively with acid fuchsin as described by Noll (1902), In the light of this work it appeared probable that while mucin formation might occur in the Golgi zone of the goblet cells as described by these authors, the origin of the granules might lie in the basal region of the cell.

scholarly journals ULTRASTRUCTURAL LOCALIZATION OF ACID MUCOSUBSTANCES IN THE MOUSE COLON WITH IRON-CONTAINING STAINS

1966 ◽  
Vol 30 (2) ◽  
pp. 299-315 ◽  
Author(s):  
Mary G. Wetzel ◽  
Bruce K. Wetzel ◽  
Samuel S. Spicer
Keyword(s):  
Ferric Chloride ◽  
Ultrastructural Localization ◽  
Cell Types ◽  
Capillary Endothelium ◽  
Colloidal Iron ◽  
Specific Staining ◽  
Mouse Colon ◽  
Specific Localization ◽  
Cryostat Sections ◽  
Acid Mucosubstances

Selective ultrastructural staining of acid mucosubstances in sites containing histochemically identifiable sulfo- and sialomucins has been obtained in fixed cryostat sections with both ferric chloride and colloidal iron solutions. The rectosigmoid region of mouse colon was fixed in glutaraldehyde, formalin, or phosphate-buffered osmium tetroxide, and 40 µ cryostat sections of this material were treated with 0.1 to 0.4% ferric chloride or with a solution of dialyzed ferric chloride, ammonia, and glycerin. Specific staining depended upon the pH of the iron-containing solutions, and the optimal value was found to be approximately 2.0. Specific localization of acid mucosubstances has been noted in intracellular sites, including globules within colonic goblet cells and "deep crypt" mucous cells, small vesicles of the superficial nongoblet epithelial cells, and Golgi lamellae within each of these cell types. Extracellular material, presumed to be acid mucosubstance, was found on the surface of the epithelial microvilli and on the lumenal surface of capillary endothelium. Similar material formed a reticular network surrounding stromal cells, collagen bundles, and various colonic connective tissue elements.

Advanced Gastric Carcinoma With a Complete Intestinal Metaplasia Phenotype Associated With Early Intestinal-Type Carcinoma

2004 ◽  
Vol 128 (2) ◽  
pp. 218-221
Author(s):  
Francisco Rivera-Hueto ◽  
Encarnación Lag-Asturiano ◽  
JoséC. Utrilla-Alcolea ◽  
Juan M. Herrerías-Gutiérrez
Keyword(s):  
Intestinal Metaplasia ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Intestinal Type ◽  
Gastric Disease ◽  
Low Grade ◽  
Phenotypic Properties ◽  
Periodic Acid Schiff ◽  
High Iron ◽  
Advanced Gastric Carcinoma

Abstract An unusual case of synchronous gastric carcinomas occurred in a 28-year-old man with a family history of gastric disease. Two tumor foci were identified: a well-differentiated advanced carcinoma with the phenotypic properties of complete intestinal metaplasia and an early intestinal-type carcinoma. Histochemical and immunohistochemical stains to demonstrate complete intestinal metaplasia, ie, Alcian blue pH 2.5/periodic acid–Schiff, high iron diamine/Alcian blue pH 2.5, CD10, and MUC2, were all positive in the advanced adenocarcinoma. Of all markers used, only high iron diamine/Alcian blue pH 2.5 and Alcian blue pH 0.5 were positive in the early carcinoma. In these cases, mistakes frequently are made during examination of endoscopic biopsies. Fortunately, the advanced adenocarcinoma was low grade (the patient has shown no signs of disease at 6 years postsurgery). Histopathologic, histochemical, and immunohistochemical findings suggest that an extensive substrate of complete intestinal metaplasia (corpus) and of complete and incomplete intestinal metaplasia (antrum) can be associated with two independent tumors with different phenotypes.

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