A method of detecting and quantifying viable cells in a sample. The method includes fluorescently staining the cells by adding a fluorescent dye into the sample or putting the sample in contact with the fluorescent dye. A quenching dye is then added to the stained sample, or the sample is put into contact with the quenching dye, at a pH different from the pH in the viable cells. The quenching dye used is permeable through the membrane of a viable cell and does not readily absorb fluorescence of the fluorescent dye at the pH in the viable cells, but absorbs the fluorescence of the fluorescent dye at the pH of the fluorescent dye. Next, the sample, now stained with the fluorescent dye and the quenching dye, is illuminated by an excitation light for the fluorescent dye at a pH different from the pH in the viable cells and the fluorescence emitted from the sample is collected and detected.
一种检测和量化样品中存活细胞的方法。该方法包括向样品中加入荧光
染料或使样品与荧光
染料接触,对细胞进行荧光染色。然后在染色样品中加入淬灭
染料,或让样品与淬灭
染料接触,淬灭
染料的 pH 值与存活细胞的 pH 值不同。所使用的淬灭
染料可透过存活细胞的膜,在存活细胞的 pH 值下不易吸收荧光
染料的荧光,但在荧光
染料的 pH 值下会吸收荧光
染料的荧光。接下来,用荧光
染料和淬灭
染料对样品进行染色,用荧光
染料的激发光照射样品,激发光的 pH 值不同于有活力细胞的 pH 值,收集并检测样品发出的荧光。