THE SPECIAL VALUE OF METHODS THAT COLOR BOTH ACIDIC AND VICINAL HYDROXYL GROUPS IN THE HISTOCHEMICAL STUDY OF MUCINS. WITH REVISED DIRECTIONS FOR THE COLLOIDAL IRON STAIN, THE USE OF ALCIAN BLUE G8X AND THEIR COMBINATIONS WITH THE PERIODIC ACID-SCHIFF REA

2008 ◽  
Vol 106 (2) ◽  
pp. 402-423 ◽  
Author(s):  
Robert W. Mowry
1973 ◽  
Vol 21 (9) ◽  
pp. 794-803 ◽  
Author(s):  
KAZUYORI YAMADA

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.


1967 ◽  
Vol 15 (8) ◽  
pp. 475-481 ◽  
Author(s):  
BENITO MONIS ◽  
HOWARD D. DORFMAN

This report deals with a histochemical characterization of transitional epithelium of the urinary tract of man. The cytoplasm of cells of the innermost, luminal layer is distinctly stained by the periodic acid-Schiff procedure. Enhanced staining of the luminal border of these cells suggests the presence of a differentiation of the luminal surface ( surface mucous coat) which is selectively stained by Alcian Blue and colloidal iron. The alcianophilic surface mucous coat is seen in the innermost layer of transitional epithelial cells of the urinary tract extending from the area cribrosa down to the bladder. This indicates the presence of an acid mucinous component. Digestion with neuraminidase according to the procedure of Spicer and Warren demonstrates the presence of N-acetylneuraminic acid (sialic acid) in the free cell surface of the luminal layer of transitional epithelium of man. There is abundant glycogen in transitional epithelium of man. A very characteristic basement membrane underlies transitional epithelium. It reacts with periodic acid-Schiff and often with Alcian Blue. The neuraminidase digestion does not alter the staining of basement membrane with Alcian Blue. This suggests the presence of a sialidase-resistant form of sialomucin or a sulfomucin in basement membrane. The presence of periodic acid-Schiff-positive, nondigestible granules closely corresponds to acid phosphatase-reactive granules, presumably lysosomes. Within the transitional epithelium there were lacunae which contained abundant sialomucins. These structures have been described in old textbooks of histology but do not seem to be a normal component of transitional epithelium. The significance of these structures is unknown.


2016 ◽  
Vol 3 (2) ◽  
pp. 77-84 ◽  
Author(s):  
Sri Wahyuni ◽  
. Zuchri ◽  
. Hamny ◽  
Muhammad Jalaluddin ◽  
I Ketut Mudite Adnyane

Penelitian ini bertujuan mengetahui sebaran karbohidrat pada usus biawak air dengan metode histokimia. Organ usus dari satu ekor biawak jantan dewasa dikoleksi melalui prosedur perfusi dan selanjutnya difiksasi dengan larutan paraformaldehid 4%. Usus biawak dibagi menjadi enam bagian, yaitu bagian I sampai VI dan selanjutnya diproses menjadi preparat histologi. Deteksi sebaran karbohidrat pada lapisan mukosa usus dilakukan dengan pewarnaan alcian blue (AB) pH 2,5 untuk karbohidrat asam dan periodic acid schiff (PAS) untuk karbohidrat netral. Hasil penelitian menunjukan bahwa sebaran karbohidrat asam ditemukan pada sel goblet pada keenam bagian usus dengan intensitas reaksi sedang (++) sampai kuat (+++), dan tidak ditemukan pada struktur usus lainnya. Sebaran karbohidrat netral ditemukan di seluruh permukaan jaringan usus dengan intensitas reaksi lemah (+) sedangkan intensitas reaksi sedang (++) sampai kuat (+++) ditemukan pada sel goblet. Jumlah sel goblet penghasil karbohidrat asam dan netral pada usus bagian I-IV lebih sedikit (+~++) dibandingkan usus bagian V-VI (+++). Berdasarkan hasil penelitian dapat disimpulkan bahwa sebaran karbohidrat asam dan netral ditemukan diseluruh bagian usus dengan intensitas pewarnaan yang bervariasi. Jumlah sel goblet yang terdeteksi menghasilkan kedua jenis karbohidrat tersebut lebih banyak ditemukan pada usus bagian kaudal.Kata kunci: jaringan usus, sel goblet, karbohidrat asam dan netral, Varanus salvator. (Histochemical Study of Intestinal Carbohydrates Distribution in the Water Monitor (Varanus salvator))The objective of this study was to elaborate the distribution of carbohydrate in intestine tissue of water monitor using histochemical method. Intestinal organ from an adult male water monitor was collected after perfused and subsequently fixed in paraformaldehyde 4%. Intestinal organ was divided to six regions e.g. I to VI and then processed to histological slides. The carbohydrate distribution on the mucosal surface of intestinal tissue was stained with alcian blue (AB) pH 2.5 for detect the acid carbohydrates and periodic acid Schiff (PAS) for the neutral carbohydrates. The results showed that the distribution of acid carbohydrates found in the goblet cells at the I to VI regions with vary intensity of staining reaction with good staining (++) to intense staining (+++), whereas other intestinal structures did not contain the acid carbohydrates. Furthermore, the distribution of neutral carbohydrates was found in the whole intestinal tissue with weak reaction (+), while good staining (++) to intense staining (+++) was appeared in the goblet cells. Additionally, the number of goblet cells containing acid and neutral carbohydrates at the I-IV region was fewer (+~++) than at V-VI region (+++). Conclusion of this study is the distribution of acid and neutral carbohydrates appeared in all regions of the intestinal tissue. The large number of goblet cells that secreting both of carbohydrate type was found at the caudal of intestinal tissue.Keywords: intestinal tissue, goblet cells, acid and neutral carbohydrates, Varanus salvator.


1958 ◽  
Vol 195 (3) ◽  
pp. 535-538 ◽  
Author(s):  
M. R. Shetlar ◽  
R. Palmer Howard ◽  
Walter Joel ◽  
Dorothy C. Bowling ◽  
Clara L. Shetlar

Studies were made of the time relationships existing between administration of parathyroid extract to rats, the elevation of serum glycoproteins and the deposition of polysaccharide and calcium in the kidney. Statistically significant elevations of seromucoid occured 18 hours after the injection of turpentine and 24 hours after the first injection of parathyroid extract. These data may indicate that the effects of parathyroid extracts and turpentine on seromucoid differ. Statistically significant elevations of the serum glycoproteins other than seromucoid occurred at 36 hours in both groups. The first histochemical changes in the kidney were noted 36 hours after the first injection of parathyroid extract. Intratubular lesions were noted which contained both calcium and material which stained with periodic-acid-Schiff reagent (PAS), colloidal iron and Alcian blue. Later (48 hr. after injection) another type of lesion which stained only with PAS and did not contain calcium was noted. Consequently, no definite evidence was found for the occurrence of a polysaccharide-containing lesion prior to calcification. Only cloudy swelling of the tubules was noted in the kidneys of rats injected with turpentine. Since the injection of turpentine resulted in elevated seromucoid and other serum glycoprotein levels, it is concluded that such elevations do not result per se in kidney damage.


1995 ◽  
Vol 32 (6) ◽  
pp. 628-634 ◽  
Author(s):  
S. Doliger ◽  
M. Delverdier ◽  
J. Moré ◽  
L. Longeart ◽  
A. Régnier ◽  
...  

A dermal mucinoisis, visualized as dermal alcianophilic material, is occasionally present in canine hypothyroidism (myxedema). Various histochemical reactions (alcian blue/periodic acid–Schiff [PAS], alcian blue at pH 2.6, alcian blue at pH 1.0, critical electrolytical concentrations with and without dimethylsulfoxide, differential hydrolysis by hyaluronidases) were performed on skin biopsies from six dogs (four females and two males ranging from 8 to 13 years) affected by hypothyroidism, all of them presenting dermal mucinosis in hematoxylin and eosin–stained sections. In these dogs, the only polysaccharide compound involved in the dermal mucinosis was hyaluronic acid. In this study, hyaluronic acid dermal deposits of hypothyroid dogs were significantly different from those of controls in subepidermal connective tissue and loose reticular connective tissue but not in periadnexal zones. We recommend the combined alcian blue/PAS reaction as a routine technique to assess dermal mucinosis in hypothyroid dogs.


2001 ◽  
Vol 125 (1) ◽  
pp. 141-145
Author(s):  
Rajal B. Shah ◽  
Min W. Lee ◽  
Alvaro A. Giraldo ◽  
Mahul B. Amin

Abstract Context.—We have observed intraluminal crystalloid morphology in seminal vesicles that is superficially similar to that seen in prostate neoplasia, but found little information on such morphology in the literature. Design.—Two hundred fifty-three prostate specimens (163 needle biopsies, 75 radical prostatectomies with prostate carcinoma, 11 prostates from autopsy, and 4 cystoprostatectomies without prostate carcinoma) were examined for seminal vesicle secretions, which were categorized as (a) dense platelike inspissated, (b) fluidlike, (c) crystalloid morphology, and (d) absent. Histochemical stains (periodic acid–Schiff with and without diastase, Alcian blue at pH 2.5, and mucicarmine) were performed to characterize the nature of secretions. Results.—Proteinaceous secretions were identified in 82% of seminal vesicles examined. Of these, 61% had predominantly dense, platelike, inspissated secretions, 15% had predominantly fluidlike secretions, and 24% had predominantly crystalloid morphology. Although in some cases the crystalloid morphology resembled that of prostatic intraluminal crystalloids, the seminal vesicle crystalloids differed in that they were invariably multiple, had curved edges, and had varied forms (elliptical, cylindrical, rodlike, and rectangular). Seventy-one percent of seminal vesicle crystalloids were associated with dense, platelike, inspissated secretions and appeared to be created by fracturing within platelike secretions. There was no relationship between seminal vesicle crystalloid morphology and associated malignancy in the prostate gland, as it was seen in 24% of cases with prostate carcinoma and 25% of cases without prostate carcinoma (P = 1.0000). Fluidlike secretions were positive for Alcian blue (pH 2.5) and mucicarmine, whereas dense platelike secretions and crystalloid morphology were negative for Alcian blue (pH 2.5) and mucicarmine. Conclusions.—Seminal vesicle secretions are fairly common and, when fluidlike, are composed of acid mucopolysaccharides. Inspissation of secretions appears to be associated with loss of acidity, presumably resulting in dense platelike secretions and crystallization. Awareness of both the crystalloid morphology in seminal vesicle tissue and the distinguishing features from prostatic crystalloids may be important while interpreting prostate needle biopsies in which seminal vesicle epithelium may be confused for prostate carcinoma because of a small acinar morphology with accompanying cytologic atypia and crystalloid morphology.


1971 ◽  
Vol 8 (5-6) ◽  
pp. 485-489 ◽  
Author(s):  
D. F. Brobst ◽  
R. Cottrell ◽  
A. Delez

Mucinous degenerative change was observed in the epithelial cells lining the renal pelvis, ureter, and urinary bladder of pigs with exudative epidermitis, coliform enteritis, hog cholera, and suppurative arthritis. Mucins were observed within transitional cells either as granular or homogenous material within vacuoles. Lakes filled with mucins also were formed as a result of the coalescence of mucin from degenerating transitional cells. The cells and lakes of mucin were stained selectively by periodic acid-Schiff, alcian blue, and colloidal iron. On the basis of the reactivity patterns with these stains the transitional epithelial cells were considered capable of producing acidic and neutral mucins.


2020 ◽  
Vol 27 (5) ◽  
pp. 53-61
Author(s):  
Abdullah Saleh Alkhamiss

Background: This study was undertaken to evaluate the preferred method (Giemsa or periodic acid Schiff-Alcian blue [PAS-AB] stains) of detecting Helicobacter pylori (H. pylori) in gastric mucosal biopsies in terms of sensitivity, specificity and applicability. To the best of my knowledge, this is the first report comparing Giemsa and PAS-AB staining for the detection of H. pylori in such biopsies. Methods: The formalin-fixed paraffin-embedded blocks of 49 gastric biopsies from different patients were collected from the archive of anatomical pathology at King Abdulaziz Medical City, National Guard, Riyadh, Saudi Arabia. From each block, three slides were prepared and analysed using the hematoxylin and eosin (H&E), Giemsa and PAS-AB stains to detect the presence/absence of H. pylori, and the results were compared in terms of sensitivity, specificity and applicability. Results: The majority of the biopsies in this study showed antrum-type gastric mucosa. Only 15 biopsies showed active gastritis, whereas the rest showed chronic gastritis. Three biopsies showed intestinal metaplasia. All were detected by PAS-AB stain, but only two-thirds were detected by H&E stain. Fifteen gastric biopsies showed H. pylori infection in general and in 13 of them, active gastritis cases were discovered. Fourteen out of these 15 H. pylori infection cases were detected by Giemsa stain, whereas only 13 cases were detected by H&E stain. PAS-AB stain showed the worst results since it demonstrated only 40% sensitivity and 67.65% specificity in H. pylori detection. Conclusion: Giemsa stain has better sensitivity and specificity in gastric H. pylori infection detection than PAS-AB. Therefore, using PAS-AB stain to detect H. pylori infection is not recommended.


1998 ◽  
Vol 95 (1) ◽  
pp. 97-106 ◽  
Author(s):  
NICOLA JORDAN ◽  
JULIA NEWTON ◽  
JEFFREY PEARSON ◽  
ADRIAN ALLEN

1.The observed thickness of the gastric mucus barrier varies widely, even appearing discontinuous, depending on the methods used. Here we describe the development and application of a modified periodic acid Schiff/Alcian Blue staining technique for use on cryostat sections of gastric mucosa. This technique for the first time enables the preservation and visualization of the full thickness of the adherent gastric mucus layer and the underlying mucosa. 2.In designing this novel method we have selected those procedures which would result in the least alteration to the mucus layer. The methods used were snap freezing, cryostat sectioning of the whole stomach followed by brief ethanol pretreatment (10 min in 100% ethanol), a prolonged staining with periodic acid Schiff/Alcian Blue (15 min and 2.5 h respectively), a gentle post-fixation (45 min paraformaldehyde vapour at 37 °C) and the use of a water-soluble mountant. 3.A continuous, adherent mucus layer was observed over the surface of the rat gastric mucosa (periodic acid Schiff/Alcian Blue stained) and human gastric antral biopsies (periodic acid Schiff stained). In the rat the mean (S.D.) mucus thickness measurements along the antrum to oesophageal axis (which was divided histologically into four regions, A to D) were: A, 166 (47) μm; B, 179 (48) μm; C, 184 (50) μm; D (the non-glandular stratified epithelium at the top of the stomach), Absent. In human gastric antral biopsies the mean (S.D.) mucus thickness was 144 (52) μm. 4.This new technique has enabled the visualization and precise measurement of thickness of the gastric mucus layer in rat and man. The adherent gastric mucus layer was observed to be continuous in the rat glandular stomach and human antrum. In validation experiments in rat the mean mucus thickness measurements were found to be twice those measured by conventional histological techniques, in which the mucus layer appeared discontinuous and patchy. However, they were within the range of thickness values seen in unfixed tissues and in the rat in vivo preparation.


1983 ◽  
Vol 31 (1) ◽  
pp. 53-61 ◽  
Author(s):  
E Ippolito ◽  
V A Pedrini ◽  
A Pedrini-Mille

Proteoglycan interaction with alcian blue at different concentrations of magnesium chloride was studied both in vitro and in histological sections of paraffin-embedded tissues. Our experiments indicate that a) proteoglycans with different contents of chondroitin sulfate and keratan sulfate, prepared under nondegradative conditions, are not distinguishable on the basis of the critical electrolyte concentrations at which staining is abolished; b) the state of aggregation of proteoglycans only very slightly affects the alcian blue affinity of the macromolecules at different concentrations of magnesium chloride; c) the interaction of proteoglycans with other components of the connective tissue matrix is an important factor in determining the strength of binding of alcian blue to the polyanionic macromolecules in histological sections. These factors should be considered in interpreting histochemical data obtained by staining tissue sections with alcian blue at different concentrations of magnesium chloride. Proteoglycans, like glycosaminoglycans, are only weakly periodic acid-Schiff-positive.


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