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    • 5. 发明申请
    • METABOLIC MONITORING OF CELLS IN A MICROPLATE READER
    • 微量读数器中细胞的代谢监测
    • WO1994003791A1
    • 1994-02-17
    • PCT/US1993007177
    • 1993-07-30
    • MOLECULAR DEVICES CORPORATIONHAFEMAN, Dean, G.CRAWFORD, Kimberly, L.SANCHEZ, Anthony, J.WADA, Henry, Garrett
    • MOLECULAR DEVICES CORPORATION
    • G01N21/31
    • C12M41/46C12M41/26G01N21/253
    • The invention disclosed is a method for monitoring the metabolism of cells retained in individual wells of a multiassay plate by measuring the rates of extracellular acidification comprising the steps of: placing the cells in a solution containing a pH buffer and an acid/base indicator (the acidic form having optimum absorbance in a first wavelength region and the basic form having optimum absorbance in a second wavelength region), heating the solution in the individual wells to a substantially uniform temperature, mixing the solution in each well, measuring the optical densities of light passing vertically through the wells at a first wavelength in the first wavelength region and a second wavelength in the second wavelength region, determining a ratio of the optical density at the first and second wavelengths in each of two, or more wells, and repeating the mixing, measuring, and determining steps to monitor kinetically a rate of change in extracellular pH caused by the cells. The use of selected buffering system-acid/base indicator pairs, which provide little or no optically-apparent change in pH with change in temperature, was found to be highly desirable for carrying out the method. Methods for composing such buffering system-acid/base indicator pairs are disclosed, together with examples of such pairs.
    • 本发明公开了一种通过测量细胞外酸化速率来监测多分析板单个孔中保留细胞代谢的方法,包括以下步骤:将细胞置于含有pH缓冲液和酸/碱指示剂的溶液中 在第一波长区域具有最佳吸光度的酸性形式和在第二波长区域中具有最佳吸光度的碱性形式),将各孔中的溶液加热至基本均匀的温度,将溶液混合在每个孔中,测量光的光密度 在第一波长区域中以第一波长垂直穿过阱并且在第二波长区域中穿过第二波长,确定两个或更多个孔中的每一个中的第一和第二波长处的光密度的比率,并重复混合 ,测量和确定动态监测由细胞引起的细胞外pH变化速率的步骤。 发现使用选择的缓冲系统 - 酸/碱指示器对,其提供很少或没有光学上明显的pH随温度变化的变化,对于进行该方法是非常需要的。 公开了组合这种缓冲系统酸/碱指示剂对的方法以及这些对的实例。