Method of optimizing equilibrium constants and fluorescence coef

Details Method of optimizing equilibrium constants and fluorescence coef Method of optimizing equilibrium constants and fluorescence coef

1994-09-20

1996-08-27

Gitomer, Ralph J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving viable micro-organism

435973, C12Q 102, G01N 3353

Patent

active

055500315

ABSTRACT:
In a method for measuring an intracellular ion concentration of the present invention, 4 or more solutions of a fluorescence probe dye and the ions to be measured, 3 or more solutions of an interfering biosubstance and the fluorescent probe dye, and 3 or more solutions of the interfering biosubstance, the fluorescent probe dye and the ions to be measured are prepared. Each solution is irradiated with excitation beams at three different wavelengths, and a fluorescence intensity generated from each solution is measured. Three kinds of equilibrium constants in equilibrium constant equations and 12 kinds of fluorescence coefficients in fluorescence intensity calculating equations are optimized in accordance with a method of successive approximation. With the optimized 15 constants, the equilibrium constant equations and the fluorescence intensity calculation equations are solved to obtain the concentration of ions to be measured. Accordingly, labor for preparing samples is decreased, and the constants are given by a method of successive approximation, which means that the constants are highly reliable, and accordingly the intracellular ion concentration measured based on such highly reliable constants can be accurate.

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