scholarly journals Tissue fixation with diimidoesters as an alternative to aldehydes. II. Cytochemical and biochemical studies of rat liver fixed with dimethylsuberimidate.

1976 ◽  
Vol 24 (9) ◽  
pp. 1000-1011 ◽  
Author(s):  
A R Hand ◽  
J R Hassell

Rat liver fixed with dimethylsuberimidate (DMS) was studied to investigate the use of diimidoesters as dixatives for light and electron microscopic cytochemistry. Paraffin sections of liver fixed with DMS at pH 9.5 were weakly stained with the ninhydrin-Schiff procedure, indicating extensive reaction of NH3+ groups with the fixative. Nuclei were strongly strained by the Feulgen procedure, with no background [corrected] reaction. In contrast, glutaraldehyde fixation resulted in a significant background reaction in the cytoplasm and nuclei in controls for the Schiff-based stains. DMS-fixed liver stained intensely for glycogen with the Periodic acid-Schiff procedure, and biochemical analysis of glycogen retention and extractability indicated that DMS retained considerably more glycogen in sections than glutaraldehyde. DMS-fixed liver incubated for thiamine pyrophosphatase activity revealed reaction product in ER cisternae, Goli saccules and bile canaliculi. Peroxisomes were strongly reactive for catalase activity after incubation in diaminobenzidine medium, and reaction product of glucose-6-phosphatase activity was considerably greater following DMS fixation than after glutaraldehyde. Biochemical studies revealed up to twice as musch residual activity of glucose-6-phosphatase after DMS fixation. These results suggest that DMS may be useful as a primary fixative for certain cytochemical procedures.

1982 ◽  
Vol 60 (11) ◽  
pp. 2768-2782 ◽  
Author(s):  
Richard L. Leino

Rodlet, mucous, and granular cells from the gills and intestines of two teleosts, Catostomus commersoni and Perca flavescens, were examined using light and electron microscopic cytochemical methods. The peripheral substance in the bipartite granules of rodlet cells was positive for carbohydrates (periodic acid – Schiff (PAS)) and proteins (e.g., coupled tetrazonium) but was uncolored by stains for lipid (Sudan black), nucleic acids (e.g., gallocyanin), and chemical groups of certain carbohydrates (e.g., alcian blue (AB), pH 1.0). The core of the granules showed positive protein but no carbohydrate, nucleic acid, or lipid staining. Electron microscopy revealed granules with (1) a protease resistant, periodic acid – silver methenamine positive periphery probably rich in glycoprotein and (2) a protease-digestible, silver methenamine negative core, perhaps of relatively pure protein. The granules were resistant to ribonuclease digestion. Mucous cells generally stained more strongly than rodlet cells for carbohydrate (PAS, AB, pH 2.5 and 1.0; periodic acid – silver methenamine) and had a greater range of staining intensities indicating a more variable carbohydrate component. Protein and lipid stains left mucus granules uncolored. Granular cells of P. flavescens were moderately PAS positive and stained strongly for protein. Granular cells of C. commersoni were strongly PAS positive and stained moderately for protein; their granules exhibited periodic acid – silver methenamine positive rims and unstained central "nucleoids."


1994 ◽  
Vol 31 (5) ◽  
pp. 546-552 ◽  
Author(s):  
T. Yanai ◽  
T. Masegi ◽  
K. Ueda ◽  
J. Manabe ◽  
M. Teranishi ◽  
...  

Mineralization of various degrees was found in the brains of 79 (59%) of 134 cynomolgus monkeys ( Macaca fascicularis). There was no age dependency in the incidence or severity, nor were there any abnormalities in growth, weight gain, or neurologic signs, although a slight sex difference was observed. The lesions, which were basophilic and intensely positive for periodic acid-Schiff or von Kossa stain, occurred in the vascular walls of the globus pallidus in two types: globoid bodies with prominent concentric lamellar structures in and around the arteriolar and venular wall (type A) and fine granules in the media of small or medium-sized arteries (type B). Electron microscopic examination revealed dense deposits in the degenerated media of small or medium-sized arteries or the thickened walls of the arterioles. X-ray microanalysis demonstrated the presence of calcium, phosphorus, iron, zine, magnesium, and aluminum.


1989 ◽  
Vol 34 (4) ◽  
pp. 640-643 ◽  
Author(s):  
H. D. M. Spapen ◽  
O. Segers ◽  
N. de Wit ◽  
A. Goossens ◽  
P. Buydens ◽  
...  

2009 ◽  
Vol 46 (3) ◽  
pp. 439-443 ◽  
Author(s):  
R. R. Quigley ◽  
K. E. Knowles ◽  
G. C. Johnson

An adult dog with ataxia and a lingual mass, previously diagnosed as protothecosis, was euthanized. At the postmortem examination, the lingual mass, regions of the lungs and hilar lymph nodes, liver, mesenteric and sublumbar lymph nodes, and spinal meninges had pronounced green discoloration. Histologically, pyogranulomatous inflammation and algal organisms were found in the tongue, spinal meninges, hilar and mesenteric lymph nodes, liver, and lung. The algae had cell walls positive for periodic acid-Schiff and cytoplasmic granules. Ultrastructurally, the algae had a well-defined cell wall, stacks of grana and thylakoid membrane, and dense bodies, typical of starch granules. The organisms were identified as Chlorella, a green alga, based on the results of histochemistical and electron microscopic examination. To the author's knowledge this is the first report of disseminated Chlorella infection and the first report in a companion animal.


2016 ◽  
Vol 24 (1) ◽  
pp. 5-14 ◽  
Author(s):  
Krisztina Somogyvári ◽  
Péter Móricz ◽  
Imre Gerlinger ◽  
László Kereskai ◽  
István Szanyi ◽  
...  

The aim of this study was to evaluate the short and medium-term effects of radiofrequency (RF) and potassium titanyl phosphate (KTP) and neodymium-yttrium-aluminum garnet (Nd:YAG) laser treatment on the inferior turbinate mucosa in a porcine model. Following randomization, the inferior turbinates were treated either with RF submucosally or with the KTP or the Nd:YAG laser on the surface under videoendoscopic control. Tissue samples were taken at the end of postoperative weeks 1 and 6, and were evaluated macroscopically and histopathologically. Scanning electron microscopy was implemented to demonstrate the morphological changes in the respiratory epithelium. Six weeks following the RF procedure, the mucosa was intact in all cases, and the volume of the inferior turbinates was reduced in the majority of the cases. Although a volume reduction occurred in both laser groups, more complications associated with the healing procedure were noted. With hematoxylin and eosin and periodic acid–Schiff staining, intact epithelium, and submucosal glands remained after the RF procedures at the end of postoperative week 6. Following the KTP-laser intervention, necrotizing sialometaplasia and cartilage destruction occurred, and squamous metaplasia was also apparent in the Nd:YAG group. In both laser groups, dilated glands with excess mucus were seen. The scanning electron microscopic findings demonstrated that cilia were present in all cases. In conclusion, the medium-term macroscopic results were similar in all 3 groups, but the postoperative complications were less following the RF procedure. RF procedure is minimally invasive due to the submucosal intervention that leads to a painless, function preserving recovery.


Tracheal mucin secretion has been measured from a segment of trachea, isolated in situ , in anaesthetized geese by a method that involves radioactive labelling of tracheal mucus glycoproteins (Gallagher et al. 1975). Goose tracheal mucus comes entirely from goblet cells, since the goose trachea does not contain submucosal mucous or serous glands, and this method has been used to investigate the nervous and pharmacological control of the mucin secretion from these epithelial goblet cells. The mucins secreted have been collected, fractionated, and chemically analysed. Intracellular mucin has been examined histochemically, and the results of electron microscopic observations of epithelial cells and nerves are presented. Acetylcholine increased tracheal mucin secretion, and this effect was completely blocked by atropine. Neither α- nor β-stimulant sympathomimetic amines affected tracheal mucin secretion. Stimulation of the peripheral cut ends of the descending oesophageal nerves increased tracheal mucin secretion and the majority of this response, approximately three-quarters, appeared to be cholinergic since this proportion was blocked by atropine. The mediator for the atropine-resistant part of the response is not known, but it appears not to be a β-adrenoreceptor stimulant since the response to nerve stimulation was unaffected by propranolol given at 34 μm intrasegmentally. Other possibilities are discussed. Atropine itself decreased the resting level of tracheal mucin secretion. The local anaesthetic, lignocaine, increased tracheal mucin secretion, while at the same time blocking the responses to acetylcholine and descending oesophageal nerve stimulation. The implications of this are discussed. The electrophoretic, gel filtration and ion-exchange properties of goose tracheal mucins showed that they represented high molecular mass, negatively charged glycoproteins which could be labelled biosynthetically with [ 35 S]sulphate, [ 3 H]- and [ 14 C]glucose. These mucins could be stained with Alcian blue or periodic acid Schiff reagent. The carbohydrate composition was unusual for an epithelial glycoprotein in that fucose was absent and mannose was present in small quantities. The monosaccharides present in larger quantity were galactose, N -acetylglucosamine, N -acetylgalactosamine and sialic acid. Histochemical analysis of tissue sections of gosling tracheas demonstrated that nearly all of the glycoprotein in epithelial goblet cells contained both sialic acid and sulphate residues. Sialated mucin was present also, but to a lesser extent, and many cells contained a mixture of sialated and sulphated mucins. The adult goose trachea had a high proportion of sialated glycoprotein. Electron microscopy showed a range of epithelial cell types and intra-epithelial nerves also. Many of the nerves had neurosecretory vesicles suggestive of motor function and some were near to goblet cells.


1981 ◽  
Vol 29 (11) ◽  
pp. 1263-1272 ◽  
Author(s):  
H Shio ◽  
P B Lazarow

The theoretical advantages of electron microscopic cytochemistry were utilized to look for evidence of possible connections between peroxisomes and the endoplasmic reticulum in rat liver. Established cytochemical procedures for catalase (peroxisomes) and glucose-6-phosphatase (endoplasmic reticulum) were carried out, and evidence was sought of diffusion of reaction products between the organelles. No such diffusion was observed: lead phosphate was found in the endoplasmic reticulum and in the nuclear envelope but not in peroxisomes; oxidized diaminobenzidine (DAB) was seen only in peroxisomes. In addition, both types of cytochemistry were carried out on the same tissue. The two kinds of reaction product could be distinguished by virtue of their different electron opacities. No mixing of the two reaction products was observed. These results do not support the hypothesis that peroxisomes and endoplasmic reticulum may be connected; rather, they support the idea that the two organelles exist as separate cellular compartments.


2018 ◽  
Vol 48 (6) ◽  
pp. 2389-2398
Author(s):  
Yongxia Zheng ◽  
Jiansheng Zhou ◽  
Xuebo Li ◽  
Guangtao Xu ◽  
Mingliang Jin ◽  
...  

Background/Aims: Liver progenitor cells (LPCs) were considered as a promising hepatocyte source of cell therapy for liver disease due to their self-renewal and differentiation capacities, while little is known about the mechanism of LPC differentiate into hepatocytes. This study aims to explore the effect of miR-382, a member of Dlk1-Dio3 microRNA cluster, during hepatic differentiation from LPCs. Methods: In this study, we used rat liver progenitor cell WB-F344 as LPC cell model and HGF as inducer to simulate the process of LPCs hepatic differentiation, then microRNAs were quantified by qPCR. Next, WB-F344 cell was transfected with miR-382 mimics, then hepatocyte cell trait was characterized by multiple experiments, including that periodic acid schiff staining and cellular uptake and excretion of indocyanine green to evaluate the hepatocellular function, qPCR and Western Blotting analysis to detect the hepatocyte-specific markers (ALB, Ttr, Apo E and AFP) and transmission electron microscopy to observe the hepatocellular morphology. Moreover, Luciferase reporter assay was used to determine whether Ezh2 is the direct target of miR-382. Results: We found that miR-382 increased gradually and was inversely correlated with the potential target, Ezh2, during WB-F344 hepatic differentiation. In addition, functional studies indicated that miR-382 increased the level of hepatocyte-specific genes. Conclusions: This study demonstrates that miR-382 may be a novel regulator of LPCs differentiation by targeting Ezh2.


1972 ◽  
Vol 20 (12) ◽  
pp. 995-1005 ◽  
Author(s):  
STERLING K. AINSWORTH ◽  
MORRIS J. KARNOVSKY ◽  
SUSUMU ITO

A simple technique is described for the ultrastructural localization of periodate-reactive mucosubstances and polysaccharides containing 1,2-glycols in thin sections of routinely fixed tissues. In this method the sugar residues are oxidized by periodic acid and the resulting aldehydes presumably reduce chelated bismuth subnitrate to metallic bismuth which then appears as a fine electron-opaque precipitate at the sites of the reducing sugars. The periodic acid-alkaline bismuth procedure provides a high resolution electron microscopic technique for demonstrating tissue sites of periodate-engendered groups very similar to the light microscopic periodic acid-Schiff reaction. The reaction can be prevented by the omission of periodate oxidation or alkaline bismuth subnitrate and by aldehyde blockage with the blocking agent, m-aminophenol. However, glycogen stains markedly without prior periodate oxidation, presumably through chelation of bismuth by hydroxyl groups. Other structures which stain without prior periodate oxidation are liver lysosomal dense bodies and, occasionally, ribosomes.


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