ABSTRACT
Metallographic methods, in which a target is visualized using a probe or antibody that deposits metal selectively at its binding site, offers many advantages for bright-field in situ hybridization (ISH) detection as well as for other labeling and detection methods. Autometallographically enhanced gold labeling procedures have demonstrated higher sensitivity than conventional enzyme chromogens. Enzyme metallography, a novel procedure in which an enzymatic probe is used to deposit metal directly from solution, has been used to develop bright-field ISH methods for HER2 gene determination in breast cancer and other biopsy specimens. It provides the highest level of sensitivity and resolution, both for visualizing endogenous gene copies in nonamplified tissues and for resolving multiple gene copies to allow copy enumeration in amplified tissues without the need for oil immersion or fluorescence optics. An automated enzyme metallography procedure, silver ISH, has been developed for use in slide-staining instruments. Metallographic staining also provides excellent results for immunohistochemistry and may be combined with other staining procedures for the simultaneous detection of more than one gene or combinations of genes and proteins.
Subject(s)
Gold Colloid/chemistry , In Situ Hybridization/methods , Nucleic Acids/chemistry , Silver Compounds/chemistry , Silver Staining/methods , Adenocarcinoma/chemistry , Adenocarcinoma/diagnosis , Adenocarcinoma/genetics , Breast Neoplasms/chemistry , Breast Neoplasms/diagnosis , Breast Neoplasms/genetics , Enzymes/chemistry , Female , Gold Colloid/immunology , Humans , Receptor, ErbB-2/analysis , Receptor, ErbB-2/genetics , Silver Compounds/immunologyABSTRACT
The purpose of this investigation was to compare the local effects of polyurethane (Tecothane) and silicone tubes with or without silver impregnation in rats. Bacterial colonization or infection of the exit site and/or tunnel were documented and interpreted. All tubes were placed subcutaneously or percutaneously in the neck of 41 Sprague-Dawley rats and guided beneath the dorsal muscles into the peritoneal cavity. The incidence of bacterial abscesses along the implanted tubes was evaluated daily. After 90 days, or earlier if sepsis developed, the animals were killed painlessly and various organs and tissues from the entry site and the catheter tunnel examined histologically. In the group where polyurethane tubes were placed percutaneously, there was no difference in the frequency of abscesses between silver-impregnated and non-impregnated tubes (5/6 with and 5/7 without silver). The only difference noted was in the group with percutaneously placed silicone tubes between those with and without silver. Abscesses only occurred in 2/4 animals in the silver group and in 5/5 animals in the control group. Histological examination showed no difference in either group between infectious and foreign body reactions. Silver particles in subcutaneous, muscle and peritoneal tissue could not be demonstrated.
