A novel method for the visualization of the in situ mucus layer in rat and man

1998 ◽  
Vol 95 (1) ◽  
pp. 97-106 ◽  
Author(s):  
NICOLA JORDAN ◽  
JULIA NEWTON ◽  
JEFFREY PEARSON ◽  
ADRIAN ALLEN
Keyword(s):  
Gastric Mucosa ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Blue Staining ◽  
Mucus Layer ◽  
Gastric Mucus ◽  
Periodic Acid Schiff ◽  
Novel Method ◽  
The Mean ◽  
Thickness Measurements

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.

scholarly journals Polysaccharide Storage Myopathy in Canine Phosphofructokinase Deficiency (Type VII Glycogen Storage Disease)

1990 ◽  
Vol 27 (1) ◽  
pp. 1-8 ◽  
Author(s):  
J. W. Harvey ◽  
M. B. Calderwood Mays ◽  
K. E. Gropp ◽  
F. J. Denaro
Keyword(s):  
Alcian Blue ◽  
Amorphous Material ◽  
Periodic Acid ◽  
Glycogen Storage ◽  
Trunk Muscles ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Periodic Acid Schiff ◽  
Deficiency Type ◽  
Hematoxylin And Eosin Stain

A severe, progressive myopathy developed in an 11-year-old, phosphofructokinase (PFK)-deficient, male, English Springer Spaniel dog. Results from a routine neurological examination were normal. Examination of histologic sections of skeletal muscle revealed large accumulations of material in some myofibers. These deposits were pale, basophilic, somewhat flocculent, and slightly granular with hematoxylin and eosin stain. Most fascicles examined in sections of limb and trunk muscles were affected to some degree, with up to 10% of muscle fibers being involved. Deposits stained strongly with periodic acid-Schiff and were resistant to digestion by alpha amylase but were removed by incubation with gamma amylase. Deposits were faintly positive with Gomori's methenamine silver technique and alcian blue (pH 2.5) and were brown-gray with Lugol's iodine solution but were negative with other stains. Based on staining characteristics, the deposits seemed to consist primarily of an amylopectin-like polysaccharide(s). Alcian blue staining (pH 2.5) was removed by treatment with neuraminidase but not with hyaluronidase, indicating that some sialic acid residues were also present. Electron microscopically, the deposits were composed of short granular filaments, small granules and amorphous material. They were not membrane bound. The morphologic appearance and staining characteristics of the deposits were remarkably similar to deposits previously described in human PFK-deficient myopathy. As expected, total PFK activities were markedly reduced when assayed in skeletal muscles of this dog. In contrast with other PFK-deficient dogs, muscle glycogen in this animal was not increased above that of normal dogs.

scholarly journals Periodic acid-Schiff and alcian blue immunohistochemistry to detect mucin in mucinous breast carcinoma

2020 ◽  
Vol 29 (1) ◽  
pp. 53-7 ◽  
Author(s):  
Primariadewi Rustamadji ◽  
Jason Wibowo ◽  
Belinda Murtani ◽  
Christy Magdalena
Keyword(s):  
Breast Carcinoma ◽  
Alcian Blue ◽  
Staining Method ◽  
Periodic Acid ◽  
Mucinous Carcinoma ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Periodic Acid Schiff ◽  
Pas Staining ◽  
Mucinous Breast Carcinoma

BACKGROUND Detection of mucins has been shown to correlate with several clinicopathological characteristics in patients. Currently, periodic acid-Schiff (PAS) and alcian blue staining methods are the histochemistry staining techniques that are frequently used to detect mucin. This study was aimed to evaluate PAS and alcian blue staining in differentiating mucin characteristics between invasive carcinoma of no special type (ICNST) with mucinous degeneration and mucinous carcinoma. METHODS This cross-sectional study of 33 cases included biopsies of mucinous breast carcinoma and ICNST with mucin degeneration that were histologically verified using hematoxylin and eosin (H&E) staining. The PAS and alcian blue staining were conducted in the Laboratory of Histochemistry, Department of Anatomical Pathology, Cipto Mangunkusumo Hospital. Data were recorded using SPSS software, version 21 (IBM Corp, USA). RESULTS There were 17 cases of ICNST with mucinous degeneration and 16 cases of mucinous carcinoma with age varied from 27 to 75 years. PAS had sensitivity of 87.5% and specificity of 41.2%. Alcian blue had sensitivity of 43.8% and specificity of 82.4%. CONCLUSIONS PAS staining method is better than the alcian blue staining method in distinguishing between ICNST with mucinous degeneration and mucinous carcinoma. In the limited setting laboratory, PAS staining alone can be considered to detect mucin.

Histologic and Histochemical Characterization of Seminal Vesicle Intraluminal Secretions

2001 ◽  
Vol 125 (1) ◽  
pp. 141-145
Author(s):  
Rajal B. Shah ◽  
Min W. Lee ◽  
Alvaro A. Giraldo ◽  
Mahul B. Amin
Keyword(s):  
Seminal Vesicle ◽  
Alcian Blue ◽  
Prostate Carcinoma ◽  
Periodic Acid ◽  
Prostate Gland ◽  
Seminal Vesicles ◽  
Periodic Acid Schiff ◽  
Associated Malignancy ◽  
Distinguishing Features

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.

Thyroid Follicular Hyperplasia Associated With Massive Extracellular Mucin Deposition

2017 ◽  
Vol 25 (6) ◽  
pp. 533-535 ◽  
Author(s):  
Francesco Nesa ◽  
Luca Poggi ◽  
Stefano Ferrero ◽  
Alessandro Del Gobbo
Keyword(s):  
Alcian Blue ◽  
Periodic Acid ◽  
The Other ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Thyroid Tumors ◽  
Stromal Compartment ◽  
Follicular Hyperplasia ◽  
Nodular Hyperplasia ◽  
Thyroid Parenchyma

Extensive extracellular mucin deposition is a rare pathological thyroid condition with 6 cases described in literature so far. We report another case of a 67-year-old woman, discussing histopathological features, and review the literature. Our findings showed a diffuse mucin deposition in the stromal compartment of thyroid parenchyma. Histochemical stainings showed positivity for Alcian blue staining, but not for periodic acid–Shiff staining. Our case is peculiar because this mucin deposition was associated with benign nodular hyperplasia, in contrast with the other 6 reports, which described the same stromal alterations associated with benign or malignant thyroid tumors.

scholarly journals Evaluation of Better Staining Method among Hematoxylin and Eosin, Giemsa and Periodic Acid Schiff-Alcian Blue for the Detection of Helicobacter pylori in Gastric Biopsies

2020 ◽  
Vol 27 (5) ◽  
pp. 53-61
Author(s):  
Abdullah Saleh Alkhamiss
Keyword(s):  
Helicobacter Pylori ◽  
Alcian Blue ◽  
National Guard ◽  
Periodic Acid ◽  
Giemsa Stain ◽  
Periodic Acid Schiff ◽  
Gastric Biopsies ◽  
Hematoxylin And Eosin ◽  
H Pylori ◽  
Sensitivity Specificity

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.

Effects of parathyroid extract. II: changes preceding renal calcification

1962 ◽  
Vol 203 (4) ◽  
pp. 676-680 ◽  
Author(s):  
Reagan H. Bradford ◽  
R. Palmer Howard ◽  
Walter Joel ◽  
Jerry Puls ◽  
M. R. Shetlar
Keyword(s):  
Serum Albumin ◽  
Periodic Acid ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Periodic Acid Schiff ◽  
Protein Ratio ◽  
Parathyroid Extract ◽  
Staining Techniques ◽  
Consistent Increase ◽  
Hematoxylin And Eosin

Parathyroid extract, a total of 860 units, has been administered to rats in small, progressively increasing doses over a period of 12 days. The effects on serum protein, total glycoprotein, glycoprotein/protein ratio, calcium, individual protein and glycoprotein fractions, and renal calcification have been presented. Kidney sections from each rat were studied by histochemical techniques for calcification, neutral polysaccharide, and acid mucopolysaccharide. The serum total glycoprotein, glycoprotein/protein ratio, and calcium were found to be elevated. The serum albumin was decreased, whereas the globulin fractions were essentially unchanged. The globulin glycoprotein hexose, mostly α1-globulin, was increased; albumin glycoprotein hexose showed a somewhat less consistent increase. A "precalcification" periodic acid-Schiff-staining intraluminal material was demonstrated in the kidney after parathyroid extract treatment for 4 days. This was followed approximately 2 days later by Alcian blue-staining material and calcification demonstrable by hematoxylin and eosin and by Kóssa staining techniques. This has been interpreted as suggesting a glycoprotein-containing lesion which precedes calcification.

Further evidence of incomplete neural control of muscle properties in cat tibialis anterior motor units

1995 ◽  
Vol 268 (2) ◽  
pp. C527-C534 ◽  
Author(s):  
G. A. Unguez ◽  
R. R. Roy ◽  
D. J. Pierotti ◽  
S. Bodine-Fowler ◽  
V. R. Edgerton
Keyword(s):  
Motor Unit ◽  
Tibialis Anterior ◽  
Neural Control ◽  
Periodic Acid ◽  
Muscle Fibers ◽  
Motor Units ◽  
Fiber Types ◽  
Muscle Properties ◽  
Periodic Acid Schiff ◽  
The Mean

To examine the influence of a motoneuron in maintaining the phenotype of the muscle fibers it innervates, myosin heavy chain (MHC) expression, succinate dehydrogenase (SDH) activity, and cross-sectional area (CSA) of a sample of fibers belonging to a motor unit were studied in the cat tibialis anterior 6 mo after the nerve branches innervating the anterior compartment were cut and sutured near the point of entry into the muscle. The mean, range, and coefficient of variation for the SDH activity and the CSA for both motor unit and non-motor unit fibers for each MHC profile and from each control and each self-reinnervated muscle studied was obtained. Eight motor units were isolated from self-reinnervated muscles using standard ventral root filament testing techniques, tested physiologically, and compared with four motor units from control muscles. Motor units from self-reinnervated muscles could be classified into the same physiological types as those found in control tibialis anterior muscles. The muscle fibers belonging to a unit were depleted of glycogen via repetitive stimulation and identified in periodic acid-Schiff-stained frozen sections. Whereas muscle fibers in control units expressed similar MHCs, each motor unit from self-reinnervated muscles contained a mixture of fiber types. In each motor unit, however, there was a predominance of fibers with the same MHC profile. The relative differences in the mean SDH activities found among fibers of different MHC profiles within a unit after self-reinnervation and those found among fibers in control muscles were similar, i.e., fast-2 < fast-1 < or = slow MHC fibers.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

Effect of pepsin on partially purified pig gastric mucus and purified mucin

1988 ◽  
Vol 66 (5) ◽  
pp. 367-373 ◽  
Author(s):  
Sum P. Lee ◽  
Jane F. Nicholls ◽  
Anthony M. Roberton ◽  
Han Z. Park
Keyword(s):  
Periodic Acid ◽  
Gastric Mucin ◽  
Low Density ◽  
Gastric Mucus ◽  
Experimental Conditions ◽  
Periodic Acid Schiff ◽  
Pepsin Digestion ◽  
Cscl Density Gradient ◽  
Peptic Digestion

Partially purified native-pig gastric mucus and purified pig gastric mucin, prepared by column chromatography and caesium chloride (CsCl) density-gradient ultracentrifugation, were subjected to pepsin digestion. The products of peptic digestion were chromatographed on Sepharose CL-2B, and fractions were assayed for carbohydrate by the periodic acid – Schiff reaction. The polymeric gastric mucin in the purified mucin samples was readily degraded by pepsin. In sharp contrast, the polymeric mucin in the partially purified mucus was relatively resistant to pepsin digestion. In 45 min, pepsin degraded 40% of the polymeric mucin in the purified samples, whereas it produced no significant degradation (<10%) in the partially purified mucus samples. In partially purified gastric mucus, treated with CsCl but not fractionated by ultracentrifugation, digestion with pepsin was also slow and incomplete. This showed that differences in susceptibility between partially purified and purified preparations are not due to the chaotropic effects of CsCl. In addition, the recombination of low-density nonmucin fractions in CsCl ultracentrifugation with the mucin also resisted pepsin digestion. Finally, we have shown that the low-density fractions in mucus exhibited a strong inhibitory effect of peptic activity in vitro. We conclude that under our experimental conditions, pepsin has little effect on partially purified mucus, and our findings indicate an inhibitor of peptic digestion is present in native gastric mucus. It is likely, but unproven, that this inhibitor is a noncovalently bound lipid present in the low-density fraction.

scholarly journals Histochemical properties of cartilage proteoglycans.

1983 ◽  
Vol 31 (1) ◽  
pp. 53-61 ◽  
Author(s):  
E Ippolito ◽  
V A Pedrini ◽  
A Pedrini-Mille
Keyword(s):  
Alcian Blue ◽  
Magnesium Chloride ◽  
Periodic Acid ◽  
Keratan Sulfate ◽  
Periodic Acid Schiff ◽  
Histological Sections ◽  
Connective Tissue Matrix ◽  
Cartilage Proteoglycans ◽  
Tissue Matrix

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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