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Method of optimizing equilibrium constants and fluorescence coefficients for measuring intracellular ion concentration using fluorescence probe dyes with triple wave length photometry

机译:使用三波长光度法使用荧光探针染料优化平衡常数和荧光系数以测量细胞内离子浓度的方法

摘要

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.
机译:在本发明的细胞内离子浓度的测定方法中,荧光探针染料和被测定离子的4种以上溶液,干扰生物物质和荧光探针染料的3种以上溶液,以及3种以上的生物碱溶液。制备干扰生物物质,荧光探针染料和待测离子。用三种不同波长的激发束照射每种溶液,并测量从每种溶液产生的荧光强度。根据逐次逼近的方法,优化了平衡常数方程中的三种平衡常数和荧光强度计算方程中的12种荧光系数。通过优化的15个常数,可以求解平衡常数方程式和荧光强度计算方程式,以获得待测离子的浓度。因此,减少了用于制备样品的劳动,并且通过逐次逼近的方法给出了常数,这意味着该常数是高度可靠的,并且因此基于这种高度可靠的常数测量的细胞内离子浓度可以是准确的。

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