The Use of One Normal Hydrochloric Acid as a Background Cleaner for Periodic Acid–Schiff Staining Technique

2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S118-S119
Author(s):  
Sina Iyiola ◽  
Aremu Adegoke ◽  
Samuel Adeleke
Keyword(s):  
Tap Water ◽  
Periodic Acid ◽  
Staining Technique ◽  
Glycogen Storage ◽  
Periodic Acid Schiff ◽  
Background Staining ◽  
Schiff Reagent ◽  
Alpha 1 Antitrypsin ◽  
Clear Differentiation ◽  
Better Than

Abstract Objectives Sulfurous rinsing was considered nonessential as contained in the published periodic acid–Schiff technique because it did not clear the background better than tap water washing. The objective of this work is to elucidate the use of 1N HCl as a better background cleaner in the periodic acid–Schiff staining technique. Methods This is an experimental research. Normal samples of antrium, kidney, spleen, muscle, liver, and appendix, as well as histopathologically diagnosed samples of glycogen storage disease of the liver, alpha-1-antitrypsin deficient liver, aspergillosis nasal cavity sample, and glomerulosclerosis samples were obtained from the pathological archive. These samples were stained with a newly purchased commercially prepared Schiff reagent and overstained commercially prepared Schiff’s solution causing nonspecific background staining and laboratory-prepared Schiff reagent. Oxidation was done with 1% periodic acid for 5 minutes, washed for 5 minutes with distilled water, and stained for 10 minutes in Schiff’s reagent, but 20-minute staining was observed for the overstained Schiff reagent, oxidized in running tap water for 15 minutes. It was then rinsed in 1N HCl for 20 seconds, washed in tap water for 2 minutes, and counterstained with hematoxylin for 2 minutes. The same procedure was done for another set of slides as standards without 1N HCl rinsing done. The slides were examined under the microscope and compared for specificity. Results Slides rinsed in 1N HCl showed a clear differentiation of strong, weak, and non-PAS-positive substances. The overall effect of these staining characteristics was more pronounced in the use of laboratory-prepared Schiff. Conclusion The use of 1N HCl as a differentiator made the PAS technique more specific in its staining characteristics, especially with the use of laboratory-prepared Schiff’s reagent, which is commonly used in developing countries owing to a high cost of commercially prepared Schiff’s solution.

Modified technique for periodic acid-Schiff staining of glycoproteins on agarose-film electrophoretograms.

1983 ◽  
Vol 29 (5) ◽  
pp. 836-839 ◽  
Author(s):  
S M Trivedi ◽  
C G Frondoza ◽  
R L Humphrey
Keyword(s):  
Analytical Procedure ◽  
Periodic Acid ◽  
Staining Technique ◽  
Plasma Cell Dyscrasia ◽  
Free Light Chains ◽  
Modified Technique ◽  
Periodic Acid Schiff ◽  
Background Staining ◽  
Ponceau S ◽  
Bence Jones Proteins

Abstract We have modified the periodic acid-Schiff staining technique for glycoproteins for use with thin agarose-film electrophoresis membranes. With this procedure, carbohydrate-rich proteins can be detected with negligible background staining and insignificant staining of nonglycoproteins such as albumin and nonglycosylated Bence Jones proteins (free light chains). On the other hand, carbohydrate-rich M components of immunoglobulins M and A in serum and in cerebrospinal fluid from patients with plasma cell dyscrasia are readily detected. The technique is two- to threefold more sensitive than Ponceau S. Glycoproteins in serum and body fluids can be determined as a routine analytical procedure.

Isolation of a pure suspension of rat proximal tubules

1981 ◽  
Vol 241 (4) ◽  
pp. F403-F411 ◽  
Author(s):  
P. Vinay ◽  
A. Gougoux ◽  
G. Lemieux
Keyword(s):  
Phosphoenolpyruvate Carboxykinase ◽  
Periodic Acid ◽  
Vital Staining ◽  
Staining Technique ◽  
Proximal Tubules ◽  
Periodic Acid Schiff ◽  
Homogeneous Population ◽  
Proximal Tubular ◽  
Percoll Density Gradient ◽  
Enzyme Location

A suspension of cortical tissue fragments prepared by collagenase digestion of renal cortex obtained from fed and chronically acidotic (NH4Cl) rats was separated into four bands on a Percoll density gradient. By microscopic examination, vital staining with trypan blue, and histologic staining technique (periodic acid-Schiff) the F4 band was shown to contain only (greater than 98%) proximal tubules, whereas the F1 band was significantly enriched (70%) with distal tubules contaminated by glomeruli and short segments of proximal tubules. Intra/extracellular ratios for PAH of 15 were measured in the F4 band and of 2 in F1 band. ATP was 1.4 and 2.8 mumol/g in the F4 and F1 bands, respectively, and was stable for at least 60 min. The proximal F4 band was shown to be gluconeogenic (L-glutamine or L-lactate 2.5 mM as substrate) and to adapt to metabolic acidosis. The distal F1 band was shown to be glycolytic (glucose 2.5 mM) with no changes with acid-base status. All fractions were shown to metabolize glutamine, but the metabolic fate of this amino acid was different in proximal and distal structures. A F4/F1 activity ratio for the proximal cytoplasmic phosphoenolpyruvate carboxykinase enzyme of 2.6 and 4.3 was observed in normal and acidotic rats, respectively. In contrast, a F4/F1 ratio of 0.13 and 0.22 was observed for the distal cytoplasmic hexokinase enzyme. This preparation, therefore, allows the metabolism of a homogeneous population of proximal tubular fragments to be studied and can be used to obtain information on enzyme location within the nephron.

scholarly journals The Structural Characteristics of Chicken Fibrinogen

1977 ◽  
Author(s):  
J. Pindyck ◽  
M. W. Mosesson ◽  
D. Bannerjee ◽  
D. Galanakis
Keyword(s):  
Gel Electrophoresis ◽  
Structural Characteristics ◽  
Periodic Acid ◽  
Factor Xiiia ◽  
Periodic Acid Schiff ◽  
Polymer Formation ◽  
Sensitive Site ◽  
Schiff Reagent ◽  
The One ◽  
Fibrinogen Molecule

The structure and subunit composition of chicken fibrinogen(ϕ) have been investigated. Dodecyl sulfate gel electrophoresis of unreduced specimens revealed a single ϕ band with a molecular weight of approximately 320,000. ϕ and fibrin specimens were also electrophoresed after reduction with dithiothreitol, and after crosslinking of unreduced specimens in the presence of Factor Xllla. Chromatographically separated S-sulfo chains were also studied after reptilase or thrombin treatment,and certain samples were stained with periodic acid Schiff reagent(PAS). Chicken Aα chains weresmaller than human Aα chains (54,500 vs.70,900, respectively) but, like mammalian Aα chains, they possessed a reptilase and thrombin sensitive site, were PAS negative,and undergo Factor XIIIa catalyzed α-polymer formation. The sizes of chicken Bβ and γ chains were nearly thesame as their mammalian counterparts, (i. e. 60,000 and 49,000 respectively) ; both types of chains were PAS positive. Chicken Bβ chains possessed a slowly reactive thrombin sensitive site apparently corresponding to the one in human ϕ; the chicken β chains, like mammalian β chains, did not undergo Factor XIIIa catalyzed cross-linking. Like mammalian γ chains, chicken γ chains could undergo Factor XIIIa catalyzed γ-γ dimerization and did not possess thrombin or reptilase sensitive sites. These findings indicate that the chicken fibrinogen molecule is composed of three pairs of disulfide-bridged chains corresponding in most respects to mammalian fibrinogen chains.

scholarly journals IMMUNOLOGIC STUDIES OF HEART TISSUE

1961 ◽  
Vol 113 (1) ◽  
pp. 1-16 ◽  
Author(s):  
Melvin H. Kaplan ◽  
Frederick D. Dallenbach
Keyword(s):  
Heart Disease ◽  
Connective Tissue ◽  
Gamma Globulin ◽  
Fluorescent Antibody ◽  
Periodic Acid ◽  
Heart Tissue ◽  
Periodic Acid Schiff ◽  
Schiff Reagent ◽  
Points Of View ◽  
Rheumatic Heart

Using fluorescent antibody methods, deposits of bound gamma globulin, as determined in unfixed washed sections of auricular appendages from rheumatic hearts, were noted in a significant number (18 per cent) of 100 specimens studied. Such deposits were observed in myofibers, sarcolemma, interstitial connective tissue, and vessel walls. Albumin and fibrin were generally found absent from these sites. Control hearts from normal and pathologic material, including postmortem and biopsied specimens, in general, did not reveal such deposits. These various tissue sites which contained bound gamma globulin frequently exhibited evidence of alteration as indicated both by enhanced affinity for eosin and by strongly positive reaction with the periodic acid-Schiff reagent, and appeared comparable in some cases to "fibrinoid." Bound gamma globulin was not observed in cellular or stromal components of Aschoff lesions, nor was the occurrence of Aschoff lesions correlated with presence of bound gamma globulin. It is suggested that deposition of gamma globulin and the eosinophilic alteration associated with such deposition are related to certain of the pathologic changes of rheumatic heart disease. The nature of such deposits of gamma globulin was considered from immune and non-immune points of view.

The Properties of Soluble Proteins Synthesized by Nephrotic Rat Kidney Microsomes

1972 ◽  
Vol 50 (3) ◽  
pp. 292-298
Author(s):  
V. N. Katiyar ◽  
B. L. Dinh
Keyword(s):  
Gel Electrophoresis ◽  
Rat Kidney ◽  
Periodic Acid ◽  
Rabbit Antiserum ◽  
Periodic Acid Schiff ◽  
Rat Serum ◽  
Tissue Protein ◽  
Schiff Reagent ◽  
Normal Rat ◽  
Kidney Microsomes

The incorporation of 14C-leucine into the microsomal proteins of nephrotic rat kidney was much higher than that in the microsomal proteins of the normal rat kidney. When the newly synthesized microsomal proteins were analyzed by acrylamide gel electrophoresis, it was found that the higher incorporation of 14C-leucine in nephrotic rat kidney was mostly due to an increase in the biosynthesis of a tissue protein which migrated in the electrophoretic zone between serum albumin and transferrin. This protein did not react with rabbit antiserum to normal rat serum and was stained with periodic acid – Schiff reagent. It was believed to be excreted in the urine of nephrotic rats in great quantity and was easily identified as a distinct band in electrophoregrams of urine from these rats.

Isolation and partial characterization of a soluble mucin in human pancreatic juice

1991 ◽  
Vol 69 (8) ◽  
pp. 566-571
Author(s):  
S. P. Lee ◽  
Y. S. Choong ◽  
H. Z. Park
Keyword(s):  
Molecular Weight ◽  
Pancreatic Juice ◽  
Periodic Acid ◽  
Average Molecular Weight ◽  
Small Molecular Weight ◽  
Periodic Acid Schiff ◽  
Cellulose Acetate Electrophoresis ◽  
Cscl Density Gradient ◽  
Oligosaccharide Moiety ◽  
Schiff Reagent

Morphological and histochemical abnormalities in pancreatic mucin occur in many pancreatic disorders. However, the composition of pancreatic mucin is poorly understood. Purified mucin was isolated from pure pancreatic juice by sequential chromatography on Sepharose CL-2B and CL-4B followed by CsCl density gradient ultracentrifugation. The mucin preparation consists of 24% protein and 73% carbohydrate. Reduction of the macromolecule (> 2 × 106) by mercaptoethanol resulted in the formation of subunits of molecular weight 500 000 and released several small molecular weight proteins, including a glycoprotein of an average molecular weight of 116 000. Cellulose acetate electrophoresis separated the mucin into three species of different staining properties for periodic acid-Schiff reagent and Alcian blue, suggesting the presence of microheterogeneity with respect to sulphation and sialation. Threonine, serine, and proline composed 48% of the total amino acids, while the oligosaccharide moiety contained N-acetylglucosamine, N-acetyl-galactosamine, fucose, galactose, sialic acid, and sulphate. We also detected the presence of C16:0 and C18:0 fatty acids which were probably noncovalently bound to the pancreatic mucin.Key words: pancreas, mucin, glycoprotein, cystic fibrosis.

Human intestinal goblet cell mucin

1976 ◽  
Vol 54 (8) ◽  
pp. 707-716 ◽  
Author(s):  
I. Jabbal ◽  
D. I. C. Kells ◽  
G. Forstner ◽  
J. Forstner
Keyword(s):  
Goblet Cell ◽  
High Speed ◽  
Proteolytic Enzymes ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Strong Concentration ◽  
Acid Ratio ◽  
Schiff Reagent ◽  
Major Chemical ◽  
Sepharose 4B

Goblet cell mucin (GCM) has been purified for the first time from mucosal scrapings of human small intestine. Proteolytic enzymes and organic solvents were avoided during the isolation procedure. The mucin was purified by Sepharose 4B and 2B column chromatography of high-speed supernatant fractions. The most purified fraction was compared with rat intestinal GCM. The two were similar with respect to chemical composition, antigenic features, and polyacrylamide disc gel electrophoresis. The major chemical differences included a higher hexosamine–fucose and hexosamine – sialic acid ratio in human mucin. The two mucins showed strong concentration dependence in sedimentation velocity studies. Human mucin at a concentration of 0.2 to 1.5 mg protein per millilitre gave multiple associated peaks with variable s0 values (10.8–36.6). Rat mucin, in contrast, gave a constant (although polydisperse) pattern with s0 = 15.15. To explore these differences both mucins were stained with periodic acid – Schiff reagent and subjected to band ultracentrifugation at concentrations of 0.6–1.9 μg protein per millilitre. At this low concentration, rat mucin did not change in its sedimentation characteristics. In contrast, human GCM produced a single peak with s0 = 37.9. Thus dilution abolished polydispersity in the human but not the rat mucin, suggesting that intermolecular bonding forces in the human mucin are weaker.

scholarly journals Quantitative measurement of the histological features of alpha-1 antitrypsin deficiency-associated liver disease in biopsy specimens

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0256117
Author(s):  
George Marek ◽  
Amy Collinsworth ◽  
Chen Liu ◽  
Mark Brantly ◽  
Virginia Clark
Keyword(s):  
Liver Disease ◽  
Liver Biopsy ◽  
Periodic Acid ◽  
Therapeutic Interventions ◽  
Periodic Acid Schiff ◽  
Biopsy Specimens ◽  
Antitrypsin Deficiency ◽  
Alpha 1 Antitrypsin Deficiency ◽  
Alpha 1 Antitrypsin ◽  
Relationship Of

Background Pathological mutations in Alpha-1 Antitrypsin (AAT) protein cause retention of toxic polymers in the hepatocyte endoplasmic reticulum. The risk for cirrhosis in AAT deficiency is likely directly related to retention of these polymers within the liver. Polymers are classically identified on liver biopsy as inclusion bodies by periodic acid schiff staining after diastase treatment and immunohistochemistry. However, characterization of the polymer burden within a biopsy sample is limited to a semi-quantitative scale as described by a pathologist. Better methods to quantify polymer are needed to advance our understanding of pathogenesis of disease. Therefore, we developed a method to quantify polymer aggregation from standard histologic specimens. In addition, we sought to understand the relationship of polymer burden and other histologic findings to the presence of liver fibrosis. Methods Liver samples from a well-categorized AATD cohort were used to develop histo-morphometric tools to measure protein aggregation. Results Whole-slide morphometry reliably quantifies aggregates in AATD individuals. Despite very low levels of inclusions present (0–0.41%), accumulation of globules is not linear and is associated with higher fibrosis stages. Immunohistochemistry demonstrates that fibrosis is associated with polymer accumulation and not total AAT. A proportion of patients were found to be “heavy accumulators” with a polymer burden above the upper 25% of normal distribution. Males had significantly more liver inclusions and polymer than females. These measurements also highlight interrelated phenotypes of hepatocellular degeneration and autophagy in AATD liver disease. Conclusion Quantitative inclusion analysis measures AAT accumulation in liver biopsy specimens. Quantification of polymer may identify individuals at risk for progressive disease and candidates for therapeutic interventions. Furthermore, these methods may be useful for evaluating efficacy of drugs targeting accumulation of AAT.

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