The acridine dyes: Their purification,physicochemical, and cytochemical properties

Summary The present investigation was designed to allow a critical comparison of the cytochemical behaviour of commercially available acriflavine dye samples and pure acriflavine and proflavine dyes, regarding their application in automated cell analysis. Thin layer chromatography, NMR-spectroscopy and mass-spectrometry were applied for the identification of the dye composition.This study includes (1) a column chromatographic technique for the purification of larger dye quantities, (2) the investigation of the photodecomposition of different dye samples, and (3) the evaluation of the influence of various acriflavine/proflavine dye concentrations (1.6·10^–3–4·10^–6 mol/l) on to the emission spectrum of stained unhydrolyzed and hydrolyzed chicken erythrocytes.The commercially available acriflavine dye samples showed a much higher reduction in fluorescence intensity than the pure dyes, whereby proflavine faded less than acriflavine. Photodecomposition is markably influenced by dye impurities. Fluorescence emission spectra were registered at various acriflavine and proflavine dye concentrations for unhydrolyzed and hydrolyzed chicken erythrocytes in order to investigate the dye-dye interaction and the behaviour of the cellular DNA-dye complex. Proflavine showed a similar spectral behaviour as acriflavine. The dye concentration-dependent spectral behaviour of the DNA-dye complex of these fluorochromes seems to be a very critical factor. A comparison of quantitative fluorescence measurements can only be performed by staining cells with the same dye quality, because automated cytology requires reproducible information of cells in machinesensible terms.This investigation was supported by a grant from the Bundesministerium für Forschung und Technologie (01 VH 065)     

Stimulation der Keimung von Kiefernpollen durch UV-Bestrahlung

Zusammenfassung Bei der Aufnahme der Dosis-Effekt-Kurve für die Keimung vonPinus-Pollen nach UV-Bestrahlung in Suspension (etwa 0,4 mW/cm², 265 nm) findet man bis zu 1,5 min Bestrahlungszeit eine deutliche Stimulation des Keimvorganges.

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Stimulation of the germination of Pine Pollen by ultra-violet irradiation

Summary When investigating the dose-effect-curve of the germination of pine pollen after uv irradiation in suspension (approx. 0.4 mW/cm²,265 nm) one gets a significant stimulation of the germination process up to 1.5 min of irradiation.

     

Microfluorometric measurement at low temperature.

A cooling chamber for microfluorometric measurement is described, which allows to cool under a microscope a biological sample till 77 K and to measure it with an objective of a numerical aperture 0.6. From first experiments with BAO (5)-stained Tradescantia pollen it can be concluded, that experiments in this range of temperature open new aspects regarding the interpretation of microfluorometric phenomena.