A General-Purpose Method of Silver Staining

1955 ◽  
Vol s3-96 (35) ◽  
pp. 323-328
Author(s):  
A. PETERS
Keyword(s):  
Connective Tissue ◽  
Mercuric Chloride ◽  
Silver Staining ◽  
General Purpose ◽  
Cell Nuclei ◽  
Paraffin Sections ◽  
Nerve Fibres ◽  
Cell Cytoplasm ◽  
Ph Values ◽  
Physical Developer

A method of silver staining for paraffin sections has been described. Sections should be fixed in either Nonidez fixative, 4% formaldehyde, or 4% formaldehyde saturated with mercuric chloride. The sections are impregnated for 16 hours in 1/20,000 silver nitrate at pH 8 or 9 and developed in a glycine physical developer after the reducible silver has been removed with a 2% solution of sodium sulphite. The effect of pH on impregnation has been described. A spectrum of staining was obtained in which nerve fibres began to stain appreciably at pH 7, cell nuclei at pH 8, cell cytoplasm at pH 9, and connective tissue at higher pH values. Therefore, impregnation should be carried out at pH 8 to obtain a good staining of nerve fibres and at pH 9 if some staining of cell bodies is also required.

Detection of atrial natriuretic peptides (ANP) in rat atria by immunogold-silver staining (IGSS)

1994 ◽  
Vol 52 ◽  
pp. 304-305
Author(s):  
Robyn Rufner ◽  
Gerhard W. Hacker ◽  
Michele Forte ◽  
Nancyleigh E. Carson ◽  
Cristina Xenachis ◽  
...  
Keyword(s):  
Natriuretic Peptides ◽  
Silver Staining ◽  
Sodium Thiosulfate ◽  
Atrial Natriuretic Peptides ◽  
Paraffin Sections ◽  
Glass Slides ◽  
Metallic Salts ◽  
Wistar Kyoto ◽  
Right Atria

The use of immunogold-silver staining (IGSS) to enhance label penetration and Localization for immunocytochemistry or in situ hybridization utilizing a variety of metallic salts has been documented. In this morphological study, the effects of silver acetate, silver lactate and silver nitrate were evaluated for immunogold-labeling of a trial natriuretic peptides (ANP) in rat right atria.Eight Wistar Kyoto retired breeders were sedated with pentobarbital, perfused with either 4% paraformaldehyde (LM) or Karnovsky's fixative (EM), and right atria were dissected, processed, embedded in paraffin or epon, respectively and sectioned according to conventional methods. For light microscopy, an indirect IGSS method according to Hacker (3) was performed. Paraffin sections on glass slides were washed in ddH2O, immersed in Lugol's iodine, washed in ddH2O and treated with 2.5% aqueous sodium thiosulfate for 20 sec. After additional washes in ddH2O and TBS-0.1% fish gelatin, 10% normal goat serum (PBS with 1% BSA) was applied for 20 min before an overnight incubation at 4°C with a polyclonal α-ANP primary antibody (Peninsula Labs, 1:1000 in TBS/BSA).

scholarly journals Immunocytochemical characteristics of submucosal uterine myomas

2010 ◽  
Vol 67 (12) ◽  
pp. 977-982 ◽  
Author(s):  
Aleksandra Mladenovic-Mihailovic ◽  
Zorica Mladenovic-Bogdanovic ◽  
Predrag Mitrovic ◽  
Irena Tanaskovic ◽  
Slavica Usaj-Knezevic ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Connective Tissue ◽  
Smooth Muscle Cells ◽  
Smooth Muscle Actin ◽  
Muscle Cells ◽  
Synthetic Activity ◽  
Benign Tumors ◽  
Paraffin Sections ◽  
Weak Reaction ◽  
Synthetic Phenotype

Background/Aim. Myomas of the uterus, the most common benign tumors, have been studied for decades from the aspects of different basic and clinical disciplines. Despite this fact, their pathogenesis is still poorly understood. The aim of this study was to determine immunocytochemical characteristics of smooth muscle cells and connective tissue components of submucosal myomas of the uterus. Method. During the course of this study, 25 samples of submucosal myomas of the uterus were analyzed, all of them obtained during the surgery, after abdominal histerctomy by Aldridge. The samples were fixed in 4% formalin and embedded in paraffin. Sections of 5 ?m thickness were stained immunocytochemically using the DAKO LSAB+/HRP technique to identify ?- smooth muscle actin (?-SMA), vimentin, desmin, CD34, CD45, CD68 and PCNA (DAKO specification). Results. Our results suggest that submucosal myomas of the uterus are build-up of smooth muscle cells which are immunoreactive to ?-SMA and desmin, but also to a certain number of smooth muscle cells which are immunoreactive to ?-SMA and vimentin. Some of vimentin-immunoreactive cells also show an immunoreactivity of PCNA. In the build-up of connective stroma CD34-immunoreactive fibroblasts and neovascular formations are also present. By examining the distribution of CD45 antigen, at all the analyzed samples we observed a weak reaction. Conclusion. Submucosal myomas of the uterus are made-up of smooth muscle cells of the highly differentiated contractile phenotype (?-SMA- and desminimmunoreactivity), as well as smooth muscle cell of the synthetic phenotype which proliferate (?-SMA-, vimentin- and PCNA-immunoreactivity). In submucosal myoma of the uterus there is a significant presence of connective tissue as a result of synthetic activity of fibroblasts, which clearly differ in their immunocytochemical characteristics from smooth muscle cells of the synthetic phenotype.

scholarly journals Histomorphological changes in the rat pancreas after methionine administration

2020 ◽  
Keyword(s):  
Connective Tissue ◽  
Functional Activity ◽  
Morphological Changes ◽  
Pancreatic Tissue ◽  
European Convention ◽  
Standard Diet ◽  
Cell Nuclei ◽  
Ether Anesthesia ◽  
Rat Pancreas ◽  
Histomorphological Changes

The effectiveness of using various methionine preparations for activating pancreatic function is ambiguous; the reasons may include differences in dosage and duration of methionine administration. The question remains, in what extent the methionine application is efficacious for increasing functional activity of a healthy pancreas. The aim of our study was to investigate morphological changes in pancreas after prolonged administration of methionine. The experiments were carried out on 24 males of Wistar rats at the age of 15 months. During 21 days, the experimental animals received methionine at a daily dose of 250 mg/kg of body weight in addition to the standard diet. Histological preparations were made from pancreatic tissue according to standard method. Morphometry was performed using the computer program «Image J». The rats were taken out of the experiment under ether anesthesia. The studies were carried out in accordance with the provisions of the "European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes" (Strasbourg, 1986). Upon completion of the experiment, histomorphological sings of an increase in functional activity were registered in both exocrine (enlarged acini’s areas and their epithelium height, higher nuclear-cytoplasmic ratio of exocrinocytes, and higher number of nucleoli in cell nuclei) and endocrine (enlarged sizes of the Langerhans islets and increased number of endocrinocytes in the islets) parts of the rat pancreas. In the experimental rats, the relative area of ​​the connective tissue and the stromal-parenchyma index of the pancreas, as well as the width of the interlobular and interacinus layers of connective tissue decreased. A decrease in the mass of connective tissue in the pancreas can be considered as one of the signs of its function activation, an improvement in metabolism between acini, and an increase in regenerative capabilities. Thus, additional administration of prophylactic doses of methionine to healthy animals results in distinct morphological signs of increased pancreatic activity.

A Study of Retinal Development in Larval Herring and Sole

1980 ◽  
Vol 60 (1) ◽  
pp. 59-71 ◽  
Author(s):  
Jennifer M. Sandy ◽  
J. H. S. Blaxter
Keyword(s):  
Outer Layer ◽  
Light Microscope ◽  
Retinal Development ◽  
Visual Cell ◽  
Adult Fish ◽  
Cell Nuclei ◽  
Larval Life ◽  
Paraffin Sections ◽  
Early Stages

It is known from earlier work (Blaxter & Jones, 1967; Blaxter, 1968a; Blaxter & Staines, 1970) that the larvae of many species of teleost have a pure-cone retina. The rods only appear well after hatching and often as late as metamorphosis. This means that much of the larval life may be spent without fully developed eyes. The rods are difficult to resolve under the light microscope using paraffin sections (about 5 µm) and it is therefore easier to compare the number of cones with the prominent visual cell nuclei in the outer layer (ONL) in such preparations. The ratio has been found to be 1:1 in the early stages and changes as the rods develop, reaching ratios of 1:15 or more in adult fish. Thus, for every cone there may be 14 or more rods.

The Staining of Nervous Elements by the Bodian Method

1962 ◽  
Vol s3-103 (63) ◽  
pp. 297-310
Author(s):  
TERRY WILLIAMS
Keyword(s):  
Smooth Muscle ◽  
Paraffin Sections ◽  
Nerve Fibres ◽  
End Point ◽  
Pilot Trials ◽  
Initial Ph ◽  
Muscle Nerve ◽  
Impregnation Time ◽  
Time Required

In paraffin sections of tissues containing much smooth muscle, nerve-fibres are difficult to demonstrate by silver techniques. In pilot trials Bodian's activated protargol technique proved most promising for stomach sections. From a fixed Bodian-type schedule, the effects of variation in pH, time, and temperature of impregnation were studied separately and together. These experiments provided means of determining optimal levels for some of the variables in the process. The impregnation time required for optimal staining varies according to the temperature and initial pH. Stirring, or agitation of the slides, at intervals of a few hours has been found to be a valuable aid to even staining, and if this has been done it is possible to use the stage of clearing of the fluid as an end-point indicator.

scholarly journals The neuroma-like structure of the long-time severed and isolated nerve stumps

1965 ◽  
Vol 23 (2) ◽  
pp. 91-94 ◽  
Author(s):  
Eros Abrantes Erhart ◽  
Cecil J. Rezze
Keyword(s):  
Connective Tissue ◽  
Fibrous Tissue ◽  
Nerve Fibres ◽  
Present Complex ◽  
Proximal Stump ◽  
Long Time ◽  
Distal Nerve

The long-time severed and isolated intermediate and distal nerve segments, maintained undisturbed in their connective tissue bed, completely separated from each other and from the proximal stump, are repopulated by nerve-fibres which origin is still unknown. The extremities of such nerve segments present complex nervous nodules capped by fibrous tissue, neuroma-like structures.

scholarly journals INORGANIC CATIONS IN RAT KIDNEY

1971 ◽  
Vol 50 (3) ◽  
pp. 830-839 ◽  
Author(s):  
C. J. Tandler ◽  
A. L. Kierszenbaum
Keyword(s):  
Aqueous Solution ◽  
Connective Tissue ◽  
Rat Kidney ◽  
Basement Membranes ◽  
Cell Nuclei ◽  
Inorganic Cations ◽  
Dense Granules ◽  
Distal Tubules ◽  
The Masses ◽  
Potassium Pyroantimonate

For localization of pyroantimonate-precipitable cations, rat kidney was fixed by perfusion with a saturated aqueous solution of potassium pyroantimonate (pH about 9.2, without addition of any conventional fixative). A remarkably good preservation of the tissue and cell morphology was obtained as well as a consistent and reproducible localization of the insoluble antimonate salts of magnesium, calcium, and sodium. All proximal and distal tubules and glomeruli were delimited by massive electron-opaque precipitates localized in the basement membrane and, to a lesser extent, in adjacent connective tissue. In the intraglomerular capillaries the antimonate precipitate was encountered in the basement membranes and also between the foot processes. In addition to a more or less uniform distribution in the cytoplasm and between the microvilli of the brush border, antimonate precipitates were found in all cell nuclei, mainly between the masses of condensed chromatin. The mitochondria usually contained a few large antimonate deposits which probably correspond to the so-called "dense granules" observed after conventional fixations.

The innervation of the frog’s heart I. An examination of the autonomic postganglionic nerve fibres and a comparison of autonomic and sensory ganglion cells

1970 ◽  
Vol 176 (1042) ◽  
pp. 43-54 ◽  
Keyword(s):  
Silver Staining ◽  
Ganglion Cells ◽  
Sympathetic Ganglia ◽  
Sensory Ganglion ◽  
Nerve Fibres ◽  
Dense Networks ◽  
Histochemical Methods ◽  
Muscle Innervation ◽  
Motor Cells ◽  
Postganglionic Nerve

The pattern of innervation in the frog’s heart shown by silver staining has been compared with the results of histochemical methods. The muscle innervation is a series of dense networks of nerve bundles. These have been divided into four groups according to their size and distribution. The silver methods show fewer fibres than the histochemical methods but all the methods show the same distribution of fibres. The ganglion cells in the vagal, dorsal root and sympathetic ganglia, and in the vagosympathetic trunks and heart, have been examined with silver staining and histochemical methods. It has been concluded that the vagal ganglion cells are probably sensory; that the sympathetic ganglion cells are confined to the sympathetic chain; and that all the cells in the vagosympathetic trunks and heart are parasympathetic motor cells. Section of the vagosympathetic trunks causes loss of all fluorescent nerve fibres from the heart, while at the sametime there are no changes in the distribution of acetylcholinesterase-containing fibres in the muscle. It is concluded that the fluorescence and acetylcholinesterase methods show respectively the sympathetic and parasympathetic postganglionic fibres. There is no evidence from this work to support the claims that some recently described and extensive plexuses of fibres on the muscle are in fact nervous in origin.

On the nervous system of Saccoglossus cambrensis (Enteropneusta)

1952 ◽  
Vol 236 (634) ◽  
pp. 315-354 ◽  
Keyword(s):  
Fine Particles ◽  
Basal Layer ◽  
Superficial Layer ◽  
Nerve Ring ◽  
Cell Nuclei ◽  
Nerve Fibres ◽  
Giant Axons ◽  
Almost Everywhere ◽  
Peristaltic Contractions ◽  
Nerve Cords

A layer of nerve fibres is present almost everywhere at the base of the epidermis. It consists of a very thin basal layer of irregularly arranged fibres, and generally a thicker, more superficial layer of orientated fibres, which forms the main nerve cords and subsidiary systems of smaller through-conducting bundles. In the proboscis there are numerous longitudinal bundles, an anterior nerve ring round the basal periphery and a nerve loop under the pre-oral ciliary organ. The neurocord appears to be a simple conducting tract. In the collar epidermis numerous bundles are formed posteriorly, connecting with the prebranchial nerve ring. In the trunk the size of the longitudinal cords and the distribution of the general plexus is related to the degree of development of the muscles and cilia. The gut is well equipped with nerve fibres anteriorly, where it is particularly muscular. Practically all the nerve-cell nuclei lie outside the plexus of nerve fibres. They are very numerous and widely distributed. A concentration of bipolar neurones (Hess’s cells) occurs at the proboscis tip. Cells regarded as sensory on histological grounds are abundant round the base of the proboscis and in the groove of the ciliary organ. Large unipolar neurones are concentrated in the neurocord, some possessing ‘giant’ axons, which run posteriorly or anteriorly. The cilia of the epidermis are the chief agents of locomotion, those of the trunk being capable of synchronized reversal. They are aided by peristaltic contractions of the longitudinal muscles, which are controlled by the main longitudinal nerve cords. Burrowing peristalsis is controlled by the dorsal nerve cord of the proboscis. Some reactions to light, to the presence of fine particles and to adrenaline are described. The proboscis is necessary for spontaneous and varied activity, but the considerable degree of co-ordination shown is not due to any localized centre but to a longitudinal reflex path involving the main nerve cords. Rapid contractions of the anterior end are probably due to the giant axons. The peculiarities of the neurocord are difficult to interpret, except as a result of degeneration and paedomorphosis. The greater part of the richly nervous epidermis may be compared with the vertebrate neural plate.

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