rhodanine and 5-(4-(dimethylamino)benzylidene)rhodanine

We report a modified copper stain that can be completed in 2 hours, rather than overnight, which is required for other stains. This is useful in the diagnosis of chronic cholestatic syndromes, Wilson's disease, Indian childhood cirrhosis, and other conditions associated with copper retention.

Topics: Alum Compounds; Borates; Copper; Hematoxylin; Histocytochemistry; Hot Temperature; Humans; Liver; Rhodanine; Staining and Labeling

Six Merino sheep were dosed orally with a 0.2 per cent solution of copper sulphate, six others were undosed controls. Liver biopsies were obtained and stained for copper by the p-dimethylaminobenzylidene rhodanine (DMABR), rubeanic acid (RA) and ferricyanide (FCN) methods for examination by light microscopy. The initial and most marked accumulations of copper were found within the hepatocytes of the centrilobular zones. Increased copper loading resulted in copper deposition extending through the midlobular to the periportal zones. The deposition of copper was unequal between hepatocytes and with increasing copper loading isolated hepatocytes became packed with copper containing granules. Copper appeared within Kupffer cells and macrophages of portal triads. The first Kupffer cells to be positively stained and the greatest number of such cells were adjacent to the central veins. Accumulation of copper was demonstrated with hepatocytes at copper concentrations equivalent to 44.3 micrograms copper g-1 liver wet weight. The FCN method provided a more satisfactory demonstration of intracellular copper than the RA technique and the latter was better than the DMABR method. However, the DMABR technique provided the clearest morphological details.

Topics: Animals; Copper; Ferricyanides; Histocytochemistry; Liver; Rhodanine; Sheep; Staining and Labeling; Thioamides

5-(4-Dimethylaminobenzylidene)rhodamine, a silver-specific dye, was used in a colorimetric assay to quantitate the autoradiographic deposition of silver onto X-ray film after exposure to sodium dodecyl sulfate-polyacrylamide gels of radiolabeled biological material. Silver grains were eluted from autoradiograms with 5 N potassium hydroxide, dissolved in nitric acid, and neutralized with 1 M Trizma Base. The concentration of silver was measured spectrophotometrically owing to the chelation properties of the dye. After corrections for background exposure were made, the silver contents of excised bands were then determined by comparison to a standard curve generated with silver nitrate. We have used this silver assay to quantitate the relative amount of each polypeptide band comprising the polyomavirus structural protein VP2 doublet. The method reported here has proven useful when densitometry is inconvenient (i.e., short distance between bands, irregular shape of bands, very faint bands) in addition to being inexpensive and simple to perform.

Topics: Autoradiography; Colorimetry; Coloring Agents; Densitometry; Molecular Weight; Polyomavirus; Rhodanine; Silver; Thiazoles; Viral Proteins; Viral Structural Proteins