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.

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.

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 Cocktail of periodic Acid–Schiff and papanicolaou: Novel staining technique for the identification of leukemic eosinophils – A pilot study

2019 ◽  
Vol 23 (3) ◽  
pp. 476
Author(s):  
Lavanya Mallika ◽  
SV Sowmya ◽  
RoopaS Rao ◽  
Dominic Augustine ◽  
VanishriC Haragannavar ◽  
...  
Keyword(s):  
Pilot Study ◽  
Periodic Acid ◽  
Staining Technique ◽  
Periodic Acid Schiff

scholarly journals ALDEHYDE-FUCHSIN FOR PITUITARY CYTOCHEMISTRY

1959 ◽  
Vol 7 (2) ◽  
pp. 98-100 ◽  
Author(s):  
HERBERT ELFTMAN
Keyword(s):  
Periodic Acid ◽  
Spectrophotometric Analysis ◽  
Alcoholic Solution ◽  
Elastic Tissue ◽  
Basic Fuchsin ◽  
Tissue Component ◽  
Periodic Acid Schiff ◽  
Golgi Bodies ◽  
Aldehyde Fuchsin ◽  
Background Staining

The component of the aldehyde-fuchsin stain which is useful for pituitary staining requires more delicate manipulation than the elastic tissue component. Manufacture of the stain for pituitary purposes may he shortened to 26 hours by subjecting a 0.5% basic fuchsin solution in 70% ethanol, 0.75% paraldehyde and 1.25% HCl to incubation at 37°C. The pH of this solution precludes background staining, allowing aldehyde-fuchsin to be followed by the periodic acid-Schiff routine. Aldehyde fuchsin may be rejuvenated by adding basic fuchsin in alcoholic solution. Oxidation of pituitary tissue allows aldehyde-fuchsin to stain not only thyrotrophs but gonadotrophs as well and accentuates the staining of the hypothalamic hormones and the Golgi bodies. Since some fixatives, such as dichromates, oxidize tissue, assay of the significance of aldehyde-fuchsin staining involves awareness of the influence of the particular fixative employed. Spectrophotometric analysis of young and old aldehyde-fuchsin stains supports the assumption that staining of the pituitary thyrotrophs may involve other constituents of the stain complex than those relied on for elastic tissue visualization.

Ultrastructure of the Parotid Gland of the Nine-Banded Armadillo

1977 ◽  
Vol 35 ◽  
pp. 640-641
Author(s):  
J. R. Ruby
Keyword(s):  
Endoplasmic Reticulum ◽  
Parotid Gland ◽  
Periodic Acid ◽  
Acinar Cells ◽  
Duct System ◽  
Parotid Glands ◽  
Dasypus Novemcinctus ◽  
Anterior Portion ◽  
Periodic Acid Schiff ◽  
Thin Sections

Parotid glands were obtained from five adult (four male and one female) armadillos (Dasypus novemcinctus) which were perfusion-fixed. The glands were located in a position similar to that of most mammals. They extended interiorly to the anterior portion of the submandibular gland.In the light microscope, it was noted that the acini were relatively small and stained strongly positive with the periodic acid-Schiff (PAS) and alcian blue techniques, confirming the earlier results of Shackleford (1). Based on these qualities and other structural criteria, these cells have been classified as seromucous (2). The duct system was well developed. There were numerous intercalated ducts and intralobular striated ducts. The striated duct cells contained large amounts of PAS-positive substance.Thin sections revealed that the acinar cells were pyramidal in shape and contained a basally placed, slightly flattened nucleus (Fig. 1). The rough endoplasmic reticulum was also at the base of the cell.

Detection of Carriers in Glanzmann's Thrombasthenia

1983 ◽  
Vol 49 (03) ◽  
pp. 182-186
Author(s):  
G T E Zonneveld ◽  
E F van Leeuwen ◽  
A Sturk ◽  
J W ten Cate
Keyword(s):  
Bleeding Time ◽  
Periodic Acid ◽  
Type I ◽  
Clot Retraction ◽  
Periodic Acid Schiff ◽  
Glanzmann’S Thrombasthenia ◽  
Aggregation Response ◽  
Glanzmann's Thrombasthenia ◽  
Sds Page ◽  
Reduced State

SummaryQuantitative glycoprotein (GP) analysis of whole platelets or platelet membranes was performed by SDS-polyacrylamide gelelectrophoresis (SDS-PAGE) and periodic acid Schiff staining in the families of two unrelated Glanzmann’s thrombasthenia (GT) patients. Each family consisted of two symptom free parents, a symptom free daughter and a GT daughter. All symptom free members had a normal bleeding time, clot retraction and platelet aggregation response to adenosine 5’-diphosphate (ADP), collagen and adrenalin. Platelet Zw* antigen was normally expressed in these subjects. GT patiens, classified as a type I and II subject, showed reduced amounts of GP lib and of GP nia. Analysis of isolated membranes in the non-reduced state, however, showed that the amount of GP Ilia was also reduced in three of the four parents, whereas one parent (of the GT type I patient) and the two unaffected daughters had normal amounts of GP Ilia. Quantitative SDS-PAGE may therefore provide a method for the detection of asymptomatic carriers in GT type I and II.

Swim bladder mycosis in farmed rainbow trout Oncorhynchus mykiss caused by Phoma herbarum and experimental verification of pathogenicity

2020 ◽  
Vol 138 ◽  
pp. 237-246 ◽  
Author(s):  
J Řehulka ◽  
A Kubátová ◽  
V Hubka
Keyword(s):  
Rainbow Trout ◽  
Histopathological Examination ◽  
Periodic Acid ◽  
Bladder Wall ◽  
Swim Bladder ◽  
Cell Reactivity ◽  
Periodic Acid Schiff ◽  
Posterior Portion ◽  
Hepatic Congestion ◽  
Phoma Herbarum

In this study, spontaneous swim bladder mycosis was documented in a farmed fingerling rainbow trout from a raceway culture system. At necropsy, the gross lesions included a thickened swim bladder wall, and the posterior portion of the swim bladder was enlarged due to massive hyperplasia of muscle. A microscopic wet mount examination of the swim bladder contents revealed abundant septate hyphae, and histopathological examination showed periodic acid-Schiff-positive mycelia in the lumen and wall of the swim bladder. Histopathological examination of the thickened posterior swim bladder revealed muscle hyperplasia with expansion by inflammatory cells. The causative agent was identified as Phoma herbarum through morphological analysis and DNA sequencing. The disease was reproduced in rainbow trout fingerlings using intraperitoneal injection of a spore suspension. Necropsy in dead and moribund fish revealed extensive congestion and haemorrhages in the serosa of visceral organs and in liver and abdominal serosanguinous fluid. Histopathological examination showed severe hepatic congestion, sinusoidal dilatation, Kupffer cell reactivity, leukostasis and degenerative changes. Fungi were disseminated to the liver, pyloric caeca, kidney, spleen and heart. Although infections caused by Phoma spp. have been repeatedly reported in fish, species identification has been hampered by extensive taxonomic changes. The results of this study confirmed the pathogenicity of P. herbarum in salmonids by using a reliably identified strain during experimental fish infection and provides new knowledge regarding the course of infection.

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