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    • 6. 发明授权
    • Process for determining the pH value in the interior of a cell
    • 确定细胞内部pH值的方法
    • US4677060A
    • 1987-06-30
    • US680078
    • 1984-12-10
    • Gunter ValetGerhard Ruhenstroth-BauerErich WunschLuis Moroder
    • Gunter ValetGerhard Ruhenstroth-BauerErich WunschLuis Moroder
    • G01N21/75C07C255/51C07C255/53C07C271/06C12Q1/00C12Q1/02C12Q1/37C12Q1/44G01N21/64G01N21/80G01N33/84C12Q1/38
    • C07C255/00G01N33/84
    • 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.
    • 描述了确定细胞内部的pH值的方法。 这是通过测量通过在溶液中孵育而被细胞吸收的荧光物质的发射来实现的。 该溶液含有所述荧光物质的化合物; 通过存在于细胞中的酶将荧光物质从细胞中的该化合物中分离出来。 选择的荧光物质是这样的,使得发射最大值的频率取决于pH值。 所选择的荧光物质的化合物是膜渗透的化合物。 荧光的测量是基于测量发射最大值的频率的原理。 适用于此目的的荧光物质是2,3-二氰基-1,4-对苯二酚。 合适的膜渗透化合物是1,4-二乙酰氧基-2,3-二氰基苯或1,4-二丁酰氧基-2,3-二氰基苯或1,4-二(叔丁氧基羰基-1-烷氧基) 二氰基苯。