ABSTRACT
The photodynamic effect of harmine and UV light was studied by measuring the number of sister-chromatid exchanges and micronuclei induced by this treatment in Allium cepa meristematic cells. A significant increase in the frequencies of both cytogenetic events was observed when proliferating cells were treated for 4 h with harmine followed by irradiation with UV light for 3 min.
Subject(s)
Alkaloids/pharmacology , Allium/radiation effects , Harmine/pharmacology , Ultraviolet Rays , Allium/drug effects , Allium/genetics , Cell Nucleus/drug effects , Cell Nucleus/ultrastructure , Mutagenicity Tests , Sister Chromatid Exchange/drug effects , Sister Chromatid Exchange/radiation effectsABSTRACT
We studied the effectiveness of harmine as a substitute for 33258 Hoechst in the fluorescence-plus-Giemsa technique, using Allium cepa chromosomes after 5-bromo-2'-deoxyuridine (BrdU) incorporation. Harmine showed a photosensitizing capacity which was somewhat higher than 33258 Hoechst and used half of the time established for the usual treatment.
Subject(s)
Alkaloids , Harmine , Histocytochemistry/methods , Allium , Azure Stains , Bisbenzimidazole , Bromodeoxyuridine/metabolism , Chromatids/metabolism , DNA/metabolism , Spectrometry, FluorescenceABSTRACT
After the application of a ruthenium red (RR) solution to smears of chicken and human blood for 15 min, thrombocyte and leucocyte nuclei showed a blue-grey colour, contrasting with the red-stained erythrocyte nuclei. Extracellular matrix in frozen sections of cartilage showed the blue-grey colour after 1 h of staining. After the application of RR for a prolonged time (24 h), goblet cell mucin, granules of salivary glands and starch granules in Epon-embedded tissues were coloured blue-grey, blue-green and brown-green respectively; although they appeared red after shorter staining times. Microspectrophotometric measurements of differentially stained structures, and correlation with the spectral behaviour of a related ruthenium compound (ruthenium violet), are presented. The formation in situ of this latter compound by interaction of RR with certain substrates and the capacity of RR to distinguish different cellular structures are discussed.
Subject(s)
Blood Platelets/cytology , Cell Nucleus/ultrastructure , Erythrocytes/cytology , Leukocytes/cytology , Ruthenium Red , Ruthenium , Animals , Chickens , Drosophila , Humans , Intestine, Large/cytology , Salivary Glands/cytology , Seeds/cytologyABSTRACT
Semithin sections from glutaraldehyde-fixed, Epon-embedded tissues were stained by aqueous solutions of pyronin Y at increasing concentrations (from 10(-6) to 10(-3) mol). Mucopolysaccharide containing structures (e.g. mucin) were found stained in orange, meanwhile the chromatin and remaining tissue components appeared in a bright pink-red color. Cytophotometric measurements showed that a metachromatic shift occurs in the mucin content from goblet cells after pyronin Y staining at 10(-3) mol. Some features of the metachromatic reactions by cationic dyes are briefly discussed.
Subject(s)
Chromatin/analysis , Mucins/analysis , Pyronine , Staining and Labeling/methods , Xanthenes , Animals , Drosophila , Epoxy Resins , Intestine, Large/ultrastructure , Mice , Salivary Glands/ultrastructure , Skin/ultrastructure , Spectrophotometry , Tongue/ultrastructureABSTRACT
Staining by ruthenium red (0.5 mg/ml in borate buffer at pH = 9.2) has been used for light and electron microscopic visualization of polyanion containing structures in sections from glutaraldehyde-fixed, epoxy-embedded tissues. This staining technique can be applied in a simple and rapid way, showing the reactive cell components with suitable resolution and contrast. Preliminary spectrophotometric studies show the correspondence in absorption characteristics of the dye which is bound to polyanions in situ or in vitro.
