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PubMed1987

A further study on nucleolar silver stained granules in some mononuclear-lymphoid bone marrow cells.

Nucleolar silver stained granules (SSGs) representing nucleolus organizer regions (NORs) were investigated in human as well as rabbit bone marrows after visualization with standardized silver reaction for non-histone nucleolar argyrophilic proteins. The results indicated that few mononuclear lymphoid blast-like cells in investigated bone marrows are characterized by large irregularly shaped nucleoli which contain a larger number of SSGs than myeloblasts or proerythroblasts as well as immature or stimulated lymphocytes. Since according to previous studies the number of nucleolar SSGs decreased in the course of the erythroid, granulocytic and lymphocytic differentiation and maturation, a possibility exists that the described mononuclear lymphoid blast-like cells are even less differentiated and immature than committed stem cells for mentioned cell lines.

Adult↗
PubMed1991

Cell proliferation in intracranial tumours: selective silver staining of nucleolar organizer regions (AgNORs). Application to surgical and experimental neuro-oncology.

A novel tool in diagnostic and experimental pathology, the AgNOR-technique, which consists of visualization of ribosomal gene activity by selective silver staining, was applied to 144 cytological specimens of human tumours of the nervous system. The number of silver-stained nucleolar organizer regions (AgNORs) was correlated with the biological behaviour of the tumours investigated; low AgNOR number were observed in benign neoplasms such as meningiomas and schwannomas and higher AgNOR numbers in glioblastomas and metastases. The mean AgNOR number per cell was 3.15 in astrocytomas, 4.5 in anaplastic astrocytomas and 5.86 in glioblastoma multiforme. Benign and malignant lesions showed different distribution patterns of AgNORs, with few but centrally located AgNORs in benign, and multiple but scattered AgNORs in malignant tumours. AgNOR number per cell and AgNOR area revealed an inverse relationship (correlation coefficient -0.15, linear regression). In addition to the human tumours, two N-nitroso-N-ethyl-urea (NEU) induced tumors in BD-IX rats a mixed glioma (G-XIII) and a malignant schwannoma (N-XII), were investigated. Twelve G-XIII gliomas revealed homogenous AgNOR-counts (standard error of the mean less than 10%), with absolute values between the values obtained for human glioblastomas and metastases. Seven N-XIII subcutaneously transplanted schwannomas revealed higher AgNOR values than human schwannomas, but lower than experimental gliomas. It is concluded that the AgNOR method, as a technique for visualization of ribosomal gene activity, is valuable for assessing proliferative activity and malignancy in both diagnostic and experimental neuropathology.

Animals↗
PubMed1986

Immunogold-silver staining of lymphocyte surface antigens on cells in suspension and in lymph node cryostat sections.

An immunogold-silver staining (IGSS) technique for the light microscopical detection of leucocyte cell surface antigens in cell suspensions and cryostat sections is described. The specimens were first incubated with monoclonal mouse antibodies and then with colloidal gold-labelled goat anti-mouse antibodies. They were then immersed in a physical developer, counterstained and mounted. In light microscopy, the tissue architecture and the cellular morphology were well preserved. Positive cells showed dark granules on their surface membranes. Optimal labelling conditions were determined. This method proved to be a reliable tool for the enumeration of T-cells and their subsets in peripheral blood. The dense labelling permitted the use of panoptic counterstains like May-Grünwald-Giemsa or Wright's stain. This IGSS technique was used to determine the distribution of the T- and B-cell subsets in cryostat sections of reactive lymph nodes. The sensitivity of the method was comparable with that of immunofluorescence microscopy for cell suspensions and that of the biotin-avidin-peroxidase technique for tissue sections. Immunogold-silver staining was combined with enzyme cytochemistry. In dark-field or epipolarization microscopy the labelling appeared as bright granules on a dark background. With its dense granular membrane labelling and its good morphology IGSS is an ideal method for the study of particular cell types in mixed cell suspensions. In addition, it could be a general method for the detection of cell surface antigens in all kinds of cells and tissues.

Antigens, Surface↗
PubMed1997

Localization of the genes for major ribosomal RNA on chromosomes of the house musk shrew, Suncus murinus, at meiotic and mitotic cells by fluorescence in situ hybridization and silver staining.

The genes for major ribosomal RNA were localized on chromosomes 5pter-p15, 9q64-qter, and 13q38-qter of the house musk shrew, Suncus murinus (Insectivora, Soricidae) by silver staining of mitotic metaphase and meiotic pachytene spreads and fluorescence in situ hybridization using the human 28S-RNA genes as a probe to mitotic metaphase spreads. The data presented indicate a correlation between sites of in situ hybridization and silver staining. The finding of nuclear materials in mitosis was in a good agreement with observation in meiosis: same chromosomes carried active NORs in both meiotic and mitotic cells.

Animals↗
PubMed1998

Visualization of vaginal flora in cervical smears using a modified microwave silver-staining method.

The advantage of studying the vaginal flora to determine the bacteria and fungi present in cervical smears (as opposed to cultivation of these micro-organisms) is that the micro-organisms can be observed in their natural habitat. However, they are only faintly stained by the conventional Papanicolaou method. Accordingly, contrast is weak and visualization poor. For this reason, we developed a modified microwave silver-staining method that can be performed retrospectively on stained smears. Bacteria and fungi stain distinctly black and can be studied in greater detail, and their inter-relationship can be visualized. Haematoxylin or Eosin counterstain allows us to visualize vaginal inhabitants in relation to epithelial cells. In the series presented here, we show that a modified microwave silver-staining method is well suited to studying the ecology of micro-organisms in smears taken from women presenting to their doctor with clinical symptoms. Using this staining method, we have shown that lactobacilli overgrowth is associated with symptoms.

Actinomyces↗
PubMed2004

Whole-mount silver staining of Arabidopsis and maize: a new method for staining secondary wall thickenings in tracheary elements.

Secondary wall thickenings in tracheary elements were specifically stained by incubation of Arabidopsis and maize in Silver Stain Plus (Bio-Rad) staining solution, after pretreatment with SDS and ethanol solution. Scanning electron microscopic analysis of sections of celery revealed that silver particles were deposited on the secondary wall thickenings, indicating that the staining was due to the deposition of silver through the interaction of the stain with lignin. This method is more sensitive than the acidified phloroglucinol method.

Arabidopsis↗
PubMed1984

Improved record keeping and photography of silver-stained two-dimensional electrophoretograms by way of "XRD images".

Kodak X-Ray Duplication film provides a relatively simple and inexpensive medium for producing full-size positive-image transparencies ("XRD images") from silver-stained two-dimensional polyacrylamide gel electrophoretograms. In these XRD images, the spot or band pattern of a polychromatic silver-stained gel is represented as black against a clear background, regardless of the color of the spot in the gel itself. Thus, XRD images may be overlapped with transillumination for visual gel-to-gel comparisons, may be easily subjected to scanning densitometry, and may have the exposure adjusted to provide the proper background for black-and-white photography. I describe here the details of the XRD method and the design of a new apparatus for producing XRD images in a wet-gel "contact-print" format, which improves the method.

Amniotic Fluid↗
PubMed1978

Nucleolar organizer regions in the rabbit (Oryctolagus cuniculus) as shown by silver staining.

Nucleolar organizer regions (NOR's) were demonstrated in metaphase chromosomes of the domestic rabbit. Oryctolagus cuniculus (L.) (New Zealand white strain) using silver staining. Sequential quinacrine banding and a modification of the Ag-AS silver precipitation technique with duplicate photography allowed identification of silver staining NOR's on the short arms of chromosomes 13, 16, and 20, as well as the telomeric region of the long arms of number 21 in some cells. Chromosomes 13, 16 and 20 all have subterminal to terminal centromeres, often showed satellites and secondary constrictions, and were sometimes involved in associations.

Animals↗
PubMed1999

Alpha-synuclein immunoreactive Lewy bodies and Lewy neurites in Parkinson's disease are detectable by an advanced silver-staining technique.

Immunostaining with anti-alpha-synuclein is used to detect Lewy bodies and Lewy neurites in cases of Parkinson's disease and related disorders. To prove that the result of a modern silver method is equivalent to that achieved with immunoreactions for alpha-synuclein, individual sections were successively processed using both methods. The silver-stained sections showed all of the immunoreactive Lewy bodies, and thin Lewy neurites were detected equally well by both techniques. The present study, therefore, points to the capabilities of a modern silver-staining method which is less time consuming and less expensive than immunocytochemical techniques.

Humans↗
PubMed1993

Sensitive silver-staining detection of bacterial lipopolysaccharides in polyacrylamide gels.

For sensitive detection of bacterial lipopolysaccharides in the nanogram range, three almost identical silver-staining methods are often used, which are based on ammoniacal silver solutions and an acidic developer. We modified a method used for proteins, based on neutral silver nitrate solution and an alkaline developer, for the visualization of lipopolysaccharides in polyacrylamide gels, which yields better sensitivity and is much less prone to formation of non-specific background staining than the acidic developer-based silver stains.

Electrophoresis, Polyacrylamide Gel↗
PubMedDate not supplied

The use of radioactive bacteriophage proteins as X-Y markers for silver stained two-dimensional electrophoresis gels and quantification of the patterns.

We have demonstrated the feasibility of using bacteriophage ghost proteins, tritiated by metabolic labeling, as a set of standard markers for two-dimensional gels in which the proteins are to be detected by silver staining. The results indicate that a 2.5 microgram load of phage proteins yields a reproducible silver pattern of 48 spots. The spots can also be readily identified by radioautography and radiofluorography, establishing their value as a standard constellation of markers. Quantification of these patterns by computerized densitometry indicates a general agreement between detection by silver staining and detection by radiofluorography.

Biomarkers↗
PubMed1989

Ultrasensitive polychromatic silver staining of sodium dodecyl sulfate-polyacrylamide gels with the Gelcode system using the PhastSystem Development Unit.

The Gelcode color-based silver staining system, an improved formulation based on the original publication by Sammons et al. (Electrophoresis 1981, 2, 141-147) has been adapted to automated rapid staining in the PhastSystem Development Unit. The use of elevated temperatures in the fixation, washing, staining, and stabilization steps of the protocol reduces the total time of the process from 18 h to 1 h. The limit of detection, which is at least tenfold more sensitive than the silver staining protocol recommended for the PhastSystem, corresponds to 0.05-0.1 ng of protein per band. The method is applicable to both one- and two-dimensional polyacrylamide gels.

Electrophoresis, Polyacrylamide Gel↗
PubMed1991

Fast and sensitive silver staining of DNA in polyacrylamide gels.

The photochemically derived silver stain of nucleic acids in polyacrylamide gels originally described by Merril et al. (1981, Science 211, 1437-1438) was modified to reduce unspecific background staining and increase sensitivity (down to 1 pg/mm2 band cross-section). Detection limits for double-stranded DNA fragments from HaeIII endonuclease digests of phage phi X174 were maintained despite eliminating oxidation pretreatment of fixed gels and reducing silver nitrate concentration. Preexposure to formaldehyde during silver impregnation enhanced sensitivity and the inclusion of the silver-complexing agent sodium thiosulphate in the image developer decreased background staining. Higher formaldehyde concentration during image development resulted in darker bands with good contrast. The procedure almost halves the number of steps, solutions and experimental time required and can be used for the staining of DNA fragments in polyacrylamide gels bound to a polyester backing film by controlling temperature during image development. We have applied this improved staining procedure for the routine analysis of complex DNA profiles generated by DNA amplification fingerprinting (DAF).

Bacteriophages↗
PubMed1980

Silver stained core-like structures in chinese hamster metaphase Chromosomes.

Chinese hamster metaphase chromosomes, subjected to prolonged hypotonic pretreatment and subsequently stained with ammoniacal silver, contained a darkly-stained core-like structure in each chromatid, surrounded by a halo of dispersed chromatin which was pale yellow to brown in color. The core was variable in its appearance, ranging from a continuous linear configuration to a spiral structure or a discontinuous, particulate structure. Within the centromeric regions, the cores frequently appeared more intensely stained than elsewhere in the chromosome. The nucleolus organizers also stained darkly and appeared to be attached to the core-like structures. It remains to be determined whether the cores represent a real component of metaphase chromosome structure, or whether they are artifacts resulting from abnormal chromatin aggregation arising at the time of chromosome preparation.

Animals↗
PubMed2001

[A study on pathological changes of closed diffuse brain injury in rats with H.E., esterification-silver stain and albumin immunohistochemical stain].

OBJECTIVE: To observed the pathological changes of closed diffuse brain injury in the rats died immediately and 15 min to 5 days after the injury. METHODS: H.E. staind and esterification-silver stain were applied to investigate the closed diffuse brain injury. RESULTS: In rats died immediately after the concussive injury, a number of shrunken neurons(type I change), distended neurons(type II change) and wave-like nerve fibers were identified in the brain tissue, especially in brain-stem. At 2 h and 8 h after injury, brain edema and axonal swelling appeared clearly in the cortex and white matter, especially in brain-stem. At post-traumatic 8 h and 24 h, the axonal retraction balls began to appear. The amount of neurons undergoing type I and II changes and constraction balls increased along with the survivor time. After 4 days and 5 days, brain edema alleviated, but the retraction balls and axonal swelling still existed. With Esterifica-tion silver stain, the above changes of neurons and nerve fibers were more obvious. With ABC stain, the distribution of albumin(Al) was extended from the perio-vascular area to diffuse distribution. Al positive staining were more obvious in injuried neurons and nerve fibers. CONCLUSION: The distribution of the concussive damage in the brain are coup, contra-coup and centripental.

Albumins↗
PubMed1978

Evidence for the inheritance of silver-stained nucleolus organizer regions.

The inheritance of nucleolus organizer regions (NORs) was investigated by examining the degree of silver-staining in individual acrocentric chromosomes in two successive generations. The study was undertaken in six Down's syndrome children and their respective parents. Quinacrine fluorescent polymorphisms were used to identify individual acrocentrics and to determine which of the child's acrocentrics were informative as to parental homologue of origin. Of the 66 acrocentrics in the six children, 31 were informative. The correlation between the degree of silver=staining in the child's chromosomes and the respective parental chromosomes of origin was highly significant (P less than 0.001), with a correlation coefficient of 0.90. The results suggest that the degree of Ag-AS staining is characteristic for a particular chromosome and that this characteristic is an inherited property.

Cell Nucleolus↗
PubMed1986

Modified method of silver staining of proteins in polyacrylamide gels.

The very sensitive and reliable silver staining method to visualize proteins in polyacrylamide gels described by Wray et al. (Anal. Biochem. (1981) 118, 197-203) fails when the protein sample contains nucleic acids and/or metals. By washing the polyacrylamide gels in acetic acid and repeatedly in methanol immediately following electrophoresis and then using the procedure of Wray et al., many gels otherwise unstainable may be stained with a high degree of reliability. This method allows visualization of a minute amount of proteins in samples containing high amounts of DNA and metals.

Cell Line↗