Combination Of Methylene Blue Research Articles

Microsporidia and Candida spores: their discrimination by Calcofluor, trichrome-blue and methylene-blue combination staining.

Faeces of immunocompromised patients are often contaminated with the chitin-containing spores of microsporidia and Candida, which exclude the use of the chitin-specific fluorescent brightener Calcofluor white M2R for the identification of microsporidian spores. We developed a combination staining of Calcofluor white M2R with modified trichrome-blue staining and subsequent methylene-blue incubation which permits discrimination between these two types of spores. As a basis for diagnosis, a difference in the fluorescence pattern (365-440 nm) is combined with a difference in the light microscopic staining pattern. Under fluorescence conditions microsporidia spores have a spotted, brilliant white Calcofluor fluorescence and can easily be identified, while Candida spores show a reddish purple colour. Under the light microscope microsporidian spores show a light red colour with nonstained vacuole spots or strips in contrast to the yeast spores with their red-brown colour. This combination technique offers a highly specific means for the diagnosis of microsporidia spores in faeces.

Electrostimulation of the dehydrogenase system of yeast by alternating currents

The dehydrogenase system of Saccharomyces cerevisiae was electrostimulated utilizing Helmholtz coils (average 4 mT, 60 Hz) to induce a current density of microamps per square centimetre or a field strength of the order of millivolts per centimetre. As an indicator, the reduction rate of methylene blue (MB) was used, measured by its polarographic signal instantaneously in the range of minutes. Up to now the effect of field on the reduction yield of MB has been determined depending on the following parameters: the concentration of cells in suspension, the pre-exposure time before adding MB to the cells, the temperature of the solution, the influence of the physiological state of growing cells and the concentration of phosphate buffer. Non-linear responses were detected in some of these dependences. As a general rule one can conclude that the slower the reduction rate, the higher the stimulation effect. The combination of MB with the dehydrogenase system turns out to be a method for determination of the electromagnetic field effect. This technique may also be useful for other microorganisms.

Electrostimulation of the dehydrogenase system of yeast by alternating currents

The dehydrogenase system of Saccharomyces cerevisiae was electrostimulated utilizing Helmholtz coils (average 4 mT, 60 Hz) to induce a current density of microamps per square centimetre or a field strength of the order of millivolts per centimetre. As an indicator, the reduction rate of methylene blue (MB) was used, measured by its polarographic signal instantaneously in the range of minutes. Up to now the effect of field on the reduction yield of MB has been determined depending on the following parameters: the concentration of cells in suspension, the pre-exposure time before adding MB to the cells, the temperature of the solution, the influence of the physiological state of growing cells and the concentration of phosphate buffer. Non-linear responses were detected in some of these dependences. As a general rule one can conclude that the slower the reduction rate, the higher the stimulation effect.The combination of MB with the dehydrogenase system turns out to be a method for determination of the electromagnetic field effect. This technique may also be useful for other microorganisms.