Max-planck-gesellschaft Zur F&#xf6;rderung Der Wissenschaften E. V.

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Gunter Valet

Erich Wunsch

Luis Moroder

Gerhard Ruhenstroth-Bauer

For the simultaneous quantitative determination of the blood cells or other ells, the cell blood sample is incubated with a fluorescent stain cocktail containing fluorescent dyes and calibration particles and which stains at least one characteristic of the blood cells, and then the volume and the fluorescence of the cells and particles are measured simultaneously at at least one wavelength.

Method for the simultaneous quantitative determination of cells and

A process for determining the pH value in the interior of a cell is descrd. This is accomplished by measurement of the emission of a fluorescent substance which is absorbed by the cell through incubation in a solution. The solution contains a compound of the said fluorescent substance; the fluorescent substance is separated from this compound in the cell by an enzyme present in the cell. The fluorescent substance selected is one such that the frequency of the emission maximum is dependent on the pH value. The compound of this fluorescent substance which is selected is one which is membrane penetrating. Measurement of the fluorescence is based on the principle of measurement of the frequency at which the emission maximum occurs. A fluorescent substance suitable for this purpose is 2,3-dicyano-1,4-hydroquinone. A suitable membrane penetrating compound is 1,4-diacetoxy-2,3-dicyanobenzene or 1,4-dibutyryloxy-2,3-dicyanobenzene or 1,4-di(-tert-butyloxycarbonyl-1-alanyloxy)-2,3-dicyanobenzene.

Process for determining the pH value in the interior of a cell

A process for determining the pH value in the interior of a cell is described. This is accomplished by measurement of the emission of a fluorescent substance which is absorbed by the cell through incubation in a solution. The solution contains a compound of the said fluorescent substance; the fluorescent substance is separated from this compound in the cell by an enzyme present in the cell. The fluorescent substance selected is one such that the frequency of the emission maximum is dependent on the pH value. The compound of this fluorescent substance which is selected is one which is membrane penetrating. Measurement of the fluorescence is based on the principle of measurement of the frequency at which the emission maximum occurs. A fluorescent substance suitable for this purpose is 2,3-dicyano-1,4-hydroquinone. A suitable membrane penetrating compound is 1,4-diacetoxy-2,3-dicyanobenzene or 1,4-dibutyryloxy-2,3-dicyanobenzene or 1,4-di(-tert-butyloxycarbonyl-1-alanyloxy)-2,3-dicyanobenzene.

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Munich, Germany

Gunter Valet