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PubMed1979

Silver staining of myelin by means of physical development.

For staining myelin with silver a physical development technique has been devised that can render visible the thinnest fibers in various animal species, including fishes and reptiles, even in the early phase of myelination and may be applied to both frozen and embedded materials. Its principle is as follows: Myelin can form and bind colloidal silver particles in a 0.1% ammoniacal silver nitrate solution of pH 7.5. The production of metallic silver by other tissue elements is suppressed by the sections pretreated with a 2:1 mixture of pyridine and acetic anhydride for 30 min. The colloidal silver particles bound in the myelin are enlarged to microscopic dimensions by a special physical developer.

Animals↗
PubMedDate not supplied

Microwave-stimulated immunogold silver staining.

The effect of microwave-generated energy for the demonstration of immunoglobulins in periodate lysine paraformaldehyde dichromate fixed paraffin sections of human reactive tonsil using the immunogold silver staining technique has been evaluated. Results of the study show that microwave stimulation permits incubation times of both the primary antibody and the immunogold reagent to be greatly reduced. Specific staining of equal intensity to that produced using the longer standard procedure together with minimal background staining is achieved.

Gold↗
PubMedDate not supplied

Method for three-dimensional visualization of neurodegeneration in cupric-silver stained serial rat brain slices.

The spatial distribution of neurodegeneration in brains is difficult to visualize when working from 2-D serial slices. In studies where repetitive operant behavior measurements are made over several weeks following organic solvent exposure, definitive evidence of degeneration in brain structures may have been significantly cleared by the time the tissue is prepared histologically. The only remaining evidence that injury has occurred may be nothing more than neuronal and cellular debris. By choosing stains that are specific for this type of residual and/or indicative of specific pathology, a 3-D representation of the spatial distribution of the neuronal and cellular debris fields within the organ can be highlighted and displayed. We present a method for visualizing the spatial distribution of neuronal degeneration that can result from low-level organic solvent exposure scenarios. A cupric-silver stain highly specific for neuronal degeneration is used to identify neuronal debris fields in 73 serial slices of brains of rodents that were exposed to toluene vapors. Serial brain sections stained with cupric-silver are scanned at 600 dpi using a gray-scale protocol. Using commercially available software, scans are assembled into 3-D images showing both topographical and internal anatomical details. The reassembled images are further processed into stereo pairs. Gray-scale scans are compared to the original sections to establish gray-scale ranges for healthy and damaged tissue and artifact staining.

Animals↗
PubMed1984

A rapid silver-stain procedure for use with routine electrophoresis of cerebrospinal fluid on agarose gels.

Oligoclonal banding patterns, when present in cerebrospinal fluid (CSF) electrophoresed on agarose gels, are important in the diagnosis of multiple sclerosis and other demyelinating diseases. Typically, an 80-fold concentration of 2.5 mL of CSF has been needed before the oligoclonal banding pattern could be detected with Coomassie Brilliant Blue stain. This excessive volume requirement often is a problem, particularly in children. We describe here an improved silver-stain procedure with which we routinely can detect the oligoclonal banding pattern in 1 to 3 microL of unconcentrated CSF. The procedure is inexpensive and takes less than 45 min. Our method is quick, safe, sensitive, and easily performed in the clinical laboratory.

Cerebrospinal Fluid Proteins↗
PubMed1999

Digital image analysis of silver-stained nucleolar organizer region--associated proteins in endometrial cytologic samples.

OBJECTIVE: Digital image analysis was applied to determine the number, area and size of silver-stained nucleolar organizer regions (AgNORs) in cytologic samples from curettage in normal, hyperplastic and malignant endometrium. STUDY DESIGN: Thirty-two archival cytologic smears from curettage (previously stained by the Papanicolaou method) with the histologic diagnosis (4 inactive endometrium, 5 secretion, 5 proliferation, 5 simple hyperplasia, 5 complex hyperplasia, 3 atypical hyperplasia, 5 adenocarcinoma, grade 1) were analyzed with the AgNOR technique. Count, area and size of AgNORs were analyzed in 50 cells per sample using a magnification of 1,000x. Quantitative analysis was performed on an SFORM digital imaging system. Data were analyzed with the SPSS/PC+ program. Mann-Whitney and chi 2 tests were performed. RESULTS: The average value of AgNOR count increased from normal to hyperplastic endometrium and well-differentiated adenocarcinoma. Differences were significant except between atypical hyperplasia and adenocarcinoma. Four, five and more AgNORs in 40% or more of the nuclei were found in complex and atypical hyperplasia and adenocarcinoma. Proliferation, and simple and atypical hyperplasia had similar mean values of AgNOR area. The mean total AgNOR area value increased from normal to hyperplastic had similar mean values of AgNOR area. The mean total AgNOR area value increased from normal to hyperplastic and well-differentiated adenocarcinoma. Differences were statistically significant. AgNOR size in well-differentiated adenocarcinoma was significantly different from that in normal endometrium and different grades of hyperplasia. CONCLUSION: Digital image analysis of AgNOR count, area and size enabled a distinction to be made between normal, hyperplastic and malignant endometrium.

Adenocarcinoma↗
PubMed1980

Chemical studies on the silver staining of nucleoli.

The uptake of Ag-ions by isolated nucleoli of rat liver cells was studied. Nucleolar proteins separated by electrophoresis were examined for selective silver-staining. A mechanism for preferential Ag-staining of the nucleoli is discussed.

Animals↗
PubMed1980

Silver staining of the supernumerary chromosome in the cat-eye syndrome.

A case of the Cat-eye syndrome (CES) with 47,YX, + mar is presented. Silver staining method revealed the marker chromosome to be bisatellited. This abnormal chromosome is interpreted as the product of a Robertsonian translocation between the short arm and satellites of chromosome 22 and short arm of another D-group chromosome, probably No. 13.

Anus, Imperforate↗
PubMed2000

Estimation of total ribonucleic acid quantity from dilute samples by nondenaturing electrophoresis and silver staining.

Often, the amount of RNA that can be isolated from a defined tissue is very small. A method consisting of nondenaturing polyacrylamide gel electrophoresis and silver staining is described that can be used to evaluate the concentration of very dilute RNA samples. The method is a good starting point for assays dealing with small amounts of RNA, such as semiquantitative reverse transcription polymerase chain reaction (RT-PCR), making it possible to perform parallel assays from similar amounts of total RNA when quantitation by other methods is too insensitive. The method has been used successfully to monitor the amount of total RNA isolated from rat sciatic nerve and the rat C6 glioma cell line.

Electrophoresis, Polyacrylamide Gel↗
PubMed1993

Detection and characterization of new genetic mutations in individuals heterozygous for lactate dehydrogenase-B(H) deficiency using DNA conformation polymorphism analysis and silver staining.

Human lactate dehydrogenase (LDH)--B(H) mutant genes were analyzed by polymerase chain reaction (PCR) and DNA conformation polymorphism. We used polyacrylamide gradient gel and silver staining procedures for DCP analysis, and observed abnormal migration patterns in individuals heterozygous for the LDH-B deficiency. Subsequent sequence determination of the mutant alleles consistently resulted in detection of three single base substitutions (transversions), viz., a C to A at residue "35" (GCG, Ala-->GAG, Glu), a T to G at residue "172" (TTT, Phe-->GTT, Val), and an A to T at residue "176" (ATG, Met-->TTG, Leu). Furthermore, mismatched PCR or amplification refractory mutation system was developed for the rapid screening and confirmation of these mutations. These amino acid replacements may cause conformational changes in neighboring residues; this probably affects the active site arrangement and results in the loss of enzyme activity.

Amino Acid Sequence↗
PubMed1993

Detection of dengue virus antigens in cultured cells by using protein A-gold-silver staining (pAgs) method.

A protein A-gold-silver (pAgs) staining was developed to detect dengue virus antigens in cultured cells. The method can be carried out in either newly-subcultured or monolayered cells. Dengue virus-inoculated C6/36 clone of Aedes albopictus cells and human endothelial cells appeared brown-yellowish color on the peripheral membrane of the infected cells. In many cases, the infected C6/36 cells appeared darker than that of the infected endothelial cells. The positive results from the inoculated C6/36 cells usually appeared as early as 2 days post-inoculation for types 1, 2, and 4 of dengue viruses and 3 days for the dengue 3 virus. The same batch of specimens detected by direct immunofluorescence antibody test (DFA) showed positive 4 days post-inoculation for the types 2, 3, and 4 of dengue viruses and 6 days for the dengue 1 virus. The result also showed that all pAgs-positive specimens were also DFA-positive, but not vice versa. It suggested that pAgs is not only sensitive but also specific for dengue virus detection from inoculated cultured cells.

Aedes↗
PubMed1984

Silver staining of human aqueous humor proteins resolved by gel electrophoresis.

The detection of aqueous humor proteins resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was achieved using an ultrasensitive silver staining technique. This procedure permitted the detection of more than 35 different protein bands in human aqueous humor on a one-dimensional gel. Most of the stained proteins had an apparent molecular weight greater than 45,000 daltons.

Aqueous Humor↗
PubMed2004

Silver stains distinguish tau-positive structures in corticobasal degeneration/progressive supranuclear palsy and in Alzheimer's disease--comparison between Gallyas and Campbell-Switzer methods.

Possible differences in silver-staining profiles and their relation to tau-like immunoreactivity were investigated on cortical sections from corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), Down's syndrome (DS) and Alzheimer's disease (AD). Pairs of mirror sections were double-fluorolabeled with an anti-PHF tau (AT8) antibody and thiazin red (TR), a fluorochrome that labels fibrillary structures such as neurofibrillary tangles (NFTs). Subsequently, one of the pair was stained with Gallyas method (GAL), and the other with Campbell-Switzer method (CS). Identification of the same structure on the corresponding microscopic fields enabled a comparison of four different profiles of each structure: AT8 immunoreactivity, and affinity to TR, GAL and CS. NFTs of DS/AD, containing three- and four-repeat tau, were positive for TR, GAL and CS. AT8-immunoreactive structures of CBD/PSP, containing mainly four-repeat tau, were positive for GAL, but negative for CS and TR. This discrepancy is explainable if the argyrophilia with GAL is related to deposits containing four-repeat tau, while that with CS is linked to those containing three-repeat tau. The lack of CS labeling may also be related to poor TR staining, possibly representing scarcity of fibrillary structure in CBD/PSP. The absence of CS staining is characteristic of tau-positive structures of CBD/PSP, which is readily distinguishable from NFTs of DS/AD, hence is of potential pathological and diagnostic relevance.

Aged↗
PubMed1993

[Evaluation of mucosal proliferative activity by silver stained nucleolar organizer region proteins (Ag-NOR) in gallbladder diseases].

In order to investigate the diagnostic and prognostic utility, I measured the number and area of silver stained nucleolar organizer region proteins (Ag-NOR) in sections of surgical specimens from 71 patients with gallbladder diseases. The disease consisted of cholecystolithiasis without congenital anomaly in 20 patients, congenital biliary dilatation with anomalous arrangement of pancreaticobiliary ducts in 10, cholecystolithiasis with anomalous arrangement of pancreaticobiliary ducts without congenital biliary dilatation in 4, adenoma of the gallbladder in 5, and carcinoma of the gallbladder in 36. There was a significant difference between carcinoma and benign lesions in number of Ag-NOR, indicating that it may be a useful marker in discriminating cholecystopathic malignancy. The number of Ag-NOR was closely related with depth of invasion in carcinoma of the gallbladder, and there was a significant difference between carcinoma which invaded the tunica serosa and that which did not. The number of Ag-NOR was significantly lower in papillary adenocarcinoma than in moderately differentiated adenocarcinoma or adenosquamous carcinoma, suggesting that the proliferative activity and biological malignant potential are lower in the former.

Adult↗
PubMed1991

Double lectin and immunolabelling for transmission electron microscopy: pre- and post-embedding application using the biotin-streptavidin system and colloidal gold-silver staining.

Pre- and post-embedding methods are described that can be used for consecutive localization of two intracellular cytoplasmic binding sites in cells and tissues embedded in acrylic plastic for transmission electron microscopy. Both applications make use of the biotin-streptavidin system with colloidal gold detector particles and involve silver staining of the first gold signal to a predetermined size. Silver augmentation effectively masked any free binding sites on the biotinylated molecule and on the streptavidin complex of the first labelling reaction, thereby allowing a second cycle with the same detection system. Excellent ultrastructural localization was obtained with silver lactate as the silver ion donor in the developing solution, and the enhancement treatment did not destroy or even visibly reduce target site reactivity for the subsequently applied probe. Using these methods it was possible to achieve specific double lectin and immunological labelling; they could, however, be adapted to dual or multiple-labelling procedures with any biotinylated molecules.

Animals↗
PubMed1988

Improvement and simplification of low-background silver staining of proteins by using sodium dithionite.

High sensitivity and low background, the attractive characteristics of the procedure of Blum et al., Electrophoresis 1987, 8, 93-99, for silver staining of proteins in polyacrylamide gels have been improved by sensitizing the gels with sodium dithionite instead of sodium thiosulfate and by equilibration in water after fixation and prior to sensitization. These modifications decrease the background and allow for a longer development period, which in turn increases sensitivity and color contrast. In addition, the colors of the spots are shifted toward colder tones when compared with the original method.

Animals↗
PubMed1986

Characterization of silver-stained nucleolus organizer regions (Ag-NORs) in 16 inbred strains of the Norway rat, Rattus norvegicus.

Silver-stained nucleolus organizer regions (Ag-NORs) in metaphase chromosomes of cultured fibroblasts were compared among 16 inbred strains of the Norway rat, Rattus norvegicus. Ag-NORs were located at the secondary constrictions or juxtacentromeric regions of the short arms of chromosomes 3, 11, and 12. The frequency and relative size of Ag-NORs were found to be strain-specific, providing a genetic marker system useful for characterization of inbred strains. While considerable cell-to-cell variation was observed within a given strain, the strain-specific pattern of Ag-NORs was shown to be consistent in cultured and noncultured cells obtained from different tissues of embryos, newborns, and adults, as well as in successively cultured cells examined up to the 10th subculture generation. The patterns of Ag-NORs in F1 hybrids made between some of the inbred strains were in general agreement with those expected from the parental strains; some unexpected patterns were noted in F1 hybrids of a particular cross, suggesting the possible existence of nucleolar interactions in such interstrain hybrids, although this has to be confirmed.

Age Factors↗