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At least 343 records · Page 19Linked to original sources
PubMed1998

Rapid detection of gammaT cell receptor gene rearrangements in acute lymphoblastic leukemia by electrophoresis and silver staining: implications for detection of minimal residual disease.

Minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL) was studied using polymerase chain reaction (PCR). GammaT cell receptor (TCRG) genes are ideal targets for PCR-based detection of MRD due to their molecular characteristics. Polyacrylamide gel electrophoresis (PAGE) analysis of PCR products followed by silver staining was performed for 72 children with ALL at the onset of disease. Silver staining is an effective technique to detect gene rearrangements without the use of ethidium bromide. Moreover, this method may show heteroduplex bands of a clonal nature when both TCRG alleles are rearranged. PCR products subjected to a rapid staining protocol were recovered from the gel, reamplified by a second PCR and directly sequenced. After sequencing, we identified the junctional region and obtained patient-specific probes. In more than half of the patients we detected TCRG rearrangements that were used as molecular markers for residual disease.

Child↗
PubMed1993

The interference of cysteine thiols in the detection of glycated and non-glycated proteins by modified silver staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

In the previous study a staining intensification of in vitro glycated collagen type I versus a non-glycated one after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under a modified silver staining procedure was observed (Hodny, Z., Struzinsky, R. and Deyl, Z. (1992) J. Chromatogr. 578, 53-62). While investigating the specificity of this stain to glycation product(s) on protein we have observed that a great number of proteins (e.g. bovine serum albumin) was sensitive to this stain even in a non-glycated state. It is proposed from results of the analysis of amino acid composition of these proteins that their better stainability correlates with the amount of cysteine present in the protein. Modification of SH groups by iodoacetamide (or N-ethylmaleimide) had an inhibitory effect on the staining of bovine serum albumin (and some other proteins) in its 'native' state but had no visible inhibitory effect on their staining in the glycated state. However, the positive staining response of a great number of components from cellular lysates even after iodoacetamide treatment indicates the existence of further chemical groups (either of protein or nucleic acid origin) participating in this silver staining method.

Collagen↗
PubMed1989

Role of sialic acid in selective silver staining of red cell glycophorins.

A causal relationship between the presence of sialic acid and erythrocyte sialoglycoprotein-specific silver staining (Dzandu et al. 1984) has been examined. Quantitative chemical hydrolysis of sialic acid resulted in total loss of sialoglycoprotein-specific staining suggesting a causal role for sialic acid. It is speculated that under acidic conditions, the highly nucleophilic carboxylates of sialic acid (pKa = 2.6), may determine staining specificity by functioning as primary silver-reactive centers. Because selective staining was abrogated on gel treatment with aldehydes, alkali, thiol reagents or by temperature elevation, it is inferred that the above physico-chemical variables nullified selective staining by creating additional silver nucleation sites which culminate in non-specific staining. Acid treatment of ghosts at 80 degrees C, a standard method for sialic acid hydrolysis caused very specific polypeptide scissions, presumably at aspartyl-proline bonds. Because of the high reproducibility of the membrane protein fragmentation patterns, this method should be ideal for rapid peptide mapping of complex protein mixtures without recourse to isolating and purifying the individual polypeptides.

Electrophoresis, Polyacrylamide Gel↗
PubMed1991

Variations in the occurrence of silver-staining nucleolar organizer regions (AgNORs) in non-proliferating and proliferating tissues.

Previous studies on the subject of silver-staining nucleolar organizer regions (AgNORs) as indicators of precise proliferative status of tissues have sometimes resulted in ambiguity. The studies, however, have most frequently addressed themselves to the prognosis of neoplasias, with the aim of using AgNORs principally to distinguish between benign and malignant tumours. This investigation was to determine a base-line relationship of AgNOR clusters to proliferation and thus concentrated on normally proliferative tissues and conditionally renewing tissues after appropriate stimulation. Two murine transplantable tumours were also examined as examples of frank malignancy. As an example of the former, variations in AgNOR clusters were noted in the small intestine of man, mouse, and rat. The conditionally renewing systems of liver, prostate, and salivary glands were stimulated into proliferation by two-thirds partial hepatectomy, castration followed by treatment with testosterone, and isoproterenol treatment, respectively, in rat models; the murine sarcoma SaF and carcinoma CaNT provided relatively simple malignant tumours for AgNOR investigation. Proliferation was monitored by noting labelling indices after injection with bromodeoxyuridine (BrdUrd) in vivo followed by immunocytochemical visualization of S-phase cells. In all tissues, an increase in the size of AgNOR clusters rather than their number correlated positively with elevated labelling, particularly with the emergence of silver-staining regions of 2-3 microns visible diameter. Thus, increased AgNOR cluster size (diameter) as representative of AgNOR cluster/nucleolus volume was found to be dependent on proliferative activity in a range of normal and neoplastic tissues.

Animals↗
PubMed1998

A non-isotopic method for the detection of telomerase activity in tumour tissues: TRAP-silver staining assay.

Telomerase activity has been observed in a high proportion of specimens from a wide variety of human malignant tumours, but not in non-neoplastic tissues. Measurement of telomerase activity is a possible diagnostic maker for malignant tumours. A non-isotopic telomeric repeat amplification protocol (TRAP) silver staining method for the detection of telomerase activity in human bone tumour tissues is described. The TRAP-silver staining method was quick, safe, and effective, and can be used as a routine diagnostic method for the detection of telomerase activity in bone tumours.

Biomarkers, Tumor↗
PubMed1989

Immunogold-silver staining by capillary action.

The authors have developed an improved method for immunogold-silver staining of paraffin sections. Using a manual capillary action staining system, they were able to simplify the technical aspects of the procedure, permitting rapid processing of large batches of slides with better reproducibility. Background staining was decreased by use of buffers containing a detergent. The use of a light-stable silver reagent permitted greater control of the enhancement stage. The method yielded a high degree of contrast with negligible nonspecific staining. Sensitivity was comparable to that obtained with conventional enzymatic immunostaining. However, the authors noted that trypsinization of sections was rendered unnecessary for those antigens for which such pretreatment was usually required, and the need for special fixatives could be eliminated. The method was also applicable to immunostaining of frozen sections. Immunogold-silver staining by capillary action deserves consideration as an alternative to existing immunohistochemical methods in diagnostic histopathology.

Capillary Action↗
PubMed1997

Colloidal silver staining of electroblotted proteins for high sensitivity peptide mapping by liquid chromatography-electrospray ionization tandem mass spectrometry.

Mass spectrometric techniques for the identification of proteins either by amino acid sequencing or by correlation of mass spectral data with sequence databases are becoming increasingly sensitive and are rapidly approaching the limit of detection achieved by the staining of proteins in gels or, after electroblotting, on membranes. Here we present a technique for the sensitive staining of proteins electroblotted onto nitrocellulose or polyvinylidene difluoride membranes and enzymatic cleavage conditions for such proteins to achieve optimal recovery of peptides. The technique is based on the deposition of colloidal silver on the membrane-bound proteins. Peptide mixtures generated by proteolysis on the membrane were recovered at high yields and were compatible with analysis by reverse-phase chromatography and on-line electrospray ionization mass spectrometry. This simple and rapid colloidal silver staining procedure allowed the visualization of less than 5 ng of protein in a band and thus approached the sensitivity of silver staining in gels. We demonstrate that this method allows the detection of subpicomole amounts of electroblotted proteins and their identification by high-performance liquid chromatography-electrospray ionization tandem mass spectrometry.

Amido Black↗
PubMed1993

Demonstration of nucleolar organizer regions in lung carcinoma by silver staining.

Nucleolar organizer regions (NORs) were investigated in lung carcinomas by silver staining. This method was applied to 111 lung carcinoma specimens, including 40 with squamous cell carcinoma (SCC), 42 with adenocarcinoma (ADENO), 8 with adenosquamous carcinoma (ADESQ), 8 with small cell carcinoma (SMCC), 6 with large cell carcinoma (LGCC), and 7 with typical carcinoid tumors (CAOID). The mean AgNOR counts of ADENO, SCC, ADESQ, SMCC, and LGCC were significantly higher than those of the normal bronchial surface and those of the glandular or alveolar epithelium. The mean AgNOR count of CAOID was significantly higher than those of the normal glandular and alveolar epithelium but not that of the surface epithelium. The mean AgNOR count of SCC was significantly higher than that of bronchial squamous metaplasia, and the count of SMCC was significantly higher than that of CAOID. Within the same cancer category, the mean number of AgNORs increased in parallel with the histological tumor grades. These results indicate that the AgNOR method is useful for differentiating lung carcinoma from its normal counterparts and for evaluating histological tumor grades in the same lineage of lung carcinoma.

Adenocarcinoma↗
PubMed1982

A silver staining method for protein bands or immunoprecipitates in electrophoresis or immunoelectrophoretic treatment of unconcentrated cerebrospinal fluid proteins using the commercial agarose plates (Corning Universal System).

A highly sensitive silver staining method for protein bands or immunoprecipitates in electrophoresis or immunoelectrophoresis of unconcentrated cerebrospinal fluid using the commercial agarose plates (Corning Universal System) has been established. The new method is based on the reaction between protein-potassium ferrocyanide complex and silver nitrate as is the conventional silver staining in agar gel electrophoresis. However, some modifications were introduced into the procedures in order to obtain the densitometric values of fractionated proteins coincident with those by the usual amidoblack 10B staining for the same concentrated samples. Furthermore, in immunoelectrophoresis, several minor component proteins which could not be found by the usual staining have been detected in the cerebrospinal fluid samples.

Amido Black↗
PubMed1985

The immunogold-silver staining method. A powerful tool in histopathology.

Immunostaining of routinely fixed, wax embedded tissues may present problems to the pathologist since destruction of antigens can lead to false negative results. In an attempt to overcome this problem, we have compared the results of the standard peroxidase anti-peroxidase (PAP) method with those obtained using the newly developed and very sensitive immunogold-silver staining (IGSS) method. Sections from routine histopathological material as well as from normal tissue specimens were used in the comparison. Antisera to a variety of antigens commonly employed in pathology were used, including regulatory peptides and a range of other markers. In all cases the IGSS method was found to give superior or at least equal results to those obtained with the PAP technique. In some cases staining was obtained with IGSS method when the PAP technique gave no result. The intense black reaction product allowed much easier and more rapid screening of immunostained preparations as well as permitting sections to be counterstained with routine histological stains such as haematoxylin and eosin. It is therefore suggested that immunogold-silver staining is a valuable technique for the pathologist, particularly when examining overfixed or badly processed tissues.

Animals↗
PubMed1993

Critical analysis of the methods commonly employed in the assessment of cell proliferation: advantages of the NOR silver-staining technique in routine cyto-histopathology.

The methods commonly employed for the assessment of cell proliferation have been critically considered. Some 'historical' techniques, such as the mitotic count and the thymidine labeling index, cannot be proposed for routine pathology because of their complexity and difficult quantification. Other techniques, such as in vitro bromodeoxyuridine incorporation or in vivo bromodeoxyuridine infusion coupled with DNA flow cytometry, permit a very sophisticated analysis of the tumour cell kinetics, but can be performed only in highly specialized centers. During the past few years the immunohistochemical detection of antigens related to the cell cycle--particularly Ki67 or proliferating cell nuclear antigen (PCNA)/cyclin--has been rapidly introduced in numerous laboratories, and, at present, certainly represents the most common method for the assessment of cell proliferation in tumour samples. However, a new marker of cell proliferation has been recently described which presents technical features particularly suitable for routine use: the quantitative evaluation of interphase silver-stained nucleolar organizer regions (AgNORs). The NOR silver-staining technique is simple, very rapidly executed, inexpensive, easily quantifiable and, most important, can be performed on routinely processed cytological and histological samples.

Animals↗
PubMed1978

Fucosidosis and I-cell disease: a fine structural and silver-staining study of abnormal inclusion bodies in small-intestinal cells.

Small-intestinal cells of children with fucosidosis or the I-cell type of lysosomal storage disease were investigated with special attention to the fine structure and silver-staining patterns of abnormal inclusion bodies. The results indicate an accumulation of mucopolysaccharide and or glycoprotein, and lipid materials in the greater part of these inclusion bodies. The significantly enlarged lysosome-like bodies in the absorptive cells of these patients showed the same silver-stain affinity as the Golgi apparatus, apical vesicles and tubules, and the cell coat. This might indicate a crinophagic function of the lysosome-like bodies in the transport or secretion of cell coat material. Additional information is given on the storage of material in the significantly enlarged inclusion bodies in cultured fibroblasts and in the abnormal vacuoles of peripheral blood lymphocytes in I-cell disease.

Adolescent↗
PubMed1984

Isoelectric focusing followed by silver staining. A suitable method for routine investigation of cerebrospinal fluid proteins.

Cerebrospinal fluid (CSF) and serum samples from 361 consecutive patients were investigated. The protein pattern of each sample was examined by isoelectric focusing (IEF) of unconcentrated CSF followed by silver staining and by agar gel electrophoresis of concentrated CSF stained with amido black. 347 paired samples were compared. The results were the same in 83% of the samples. Discordant results were found in 10% of the samples. and in 7% additional information was achieved by one of the methods. IEF was superior to agar gel electrophoresis for the identification of oligoclonal immune reactions of the central nervous system. Tau globulin increase was seen in a few CSF samples, more often with IEF and then associated with other changes of the pattern. IEF of unconcentrated CSF followed by silver staining only requires 20 microliters CSF, while 3-4 ml CSF is needed for the agar gel electrophoretic method. It was concluded that IEF has several advantages compared to agar gel electrophoresis and is suitable as a routine method in clinical laboratories.

Adolescent↗
PubMed1979

Identification of a silver binding protein associated with the cytological silver staining of actively transcribing nucleolar regions.

Nucleoli isolated from Novikoff hepatoma cells were stained with AgNO3 to demonstrate the typical staining of active ribosomal cistrons. Pre-treatment of the nucleoli with 80 mM Tris-HCl (pH 7.5) -- 2.0 M NaCl did not interfere with silver staining. Treatment of the nucleoli with 80 mM Tris-HCl (pH 7.5) -- 0.15 M NaCl did, however, eliminate silver binding. Serial extraction of nucleoli with 2.0 M NaCl buffer followed by 0.15 M NaCl buffer also abolished silver staining. Analysis of the supernatant fraction of these extracts by polyacrylamide gel electrophoresis indicates that, although more than one nucleolar protein can bind silver, only one protein is associated with the staining of active ribosomal cistrons.

Animals↗
PubMedDate not supplied

A simple, reliable and inexpensive silver stain for nerve fibers in bleached skin.

A procedure for silver staining is described which leads to the selective and reliable impregnation of nerve fibers in bleached skin of vertebrates and invertebrates. In combination with osmium, the protocol enhances the staining of secondary sensory cells of mechanosensory and electrosensory organs so that the innervation pattern of each organ and the number of sensory cells per organ can easily be evaluated. The technique can be also used for staining nerve fibers in whole embryos.

Animals↗
PubMed2001

Silver stained isoelectrophoresis of tears and cerebrospinal fluid in multiple sclerosis.

Cerebrospinal fluid (CSF) analysis aids in the diagnosis of multiple sclerosis. However, this examination is invasive. The aim of this study was to assess the potentials of a new method of tears isoelectrophoresis (IEF). Silver staining of IEF was used to examined tears and CSF from 123 patients including 60 patients with multiple sclerosis (MS), 50 other neurological patients and 13 patients with inflammatory neurological diseases. Tears were collected on a Shirmer strip placed in one eye, avoiding reflex secretion. This method of IEF with silver staining allowed the detection of oligoclonal bands in tears that were truly immunoglobulin G on immunofixation. The concordance rate between tears and CSF was 83%, meaning that CSF provided no more information than tears analysis in 83% of cases. Sensitivity in tears (72%) and CSF (75%) was very close as was specificity (respectively 84 % and 86 %). High concordance between tears and CSF is the first step in developing a non invasive test which could replace lumbar puncture, particularly when this procedure is not fearalele or is refused by the patient.

Adolescent↗
PubMed1985

Silver staining as an indicator of active ribosomal genes.

Silver nitrate has been used as a cytological stain since the late 1800s. A modification of the Bielschowsky technique preferentially stains nucleoli and chromosomal nucleolus organizer regions (NORs). The specificity of staining is related to the method of preparation of the cytological specimens. The silver binds proteins and may be associated with the phosphate groups of certain phosphoproteins. Biochemical analyses of nucleolar proteins indicate that a limited array of specific proteins bind silver. A number of investigations have demonstrated that silver staining is indicative of active ribosomal RNA transcription, although a minor component may be associated with the fibrillar centers of cells in which ribosomal genes are inactive. Silver staining is a simple, reliable cytological method for the demonstration of ribosomal gene activity.

Animals↗