Measurements of absolute concentrations of NADH in cells using the phasor FLIM method
Measurements of absolute concentrations of NADH in cells using the phasor FLIM method
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DOI：
10.1364/boe.7.002441
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发表时间：
2016-07-01
影响因子：
3.4
通讯作者：
Gratton, Enrico
中科院分区：
文献类型：
--
作者：
Ma, Ning;Digman, Michelle A.;Gratton, Enrico
我们提出了一种使用荧光衰减的相量表示来推导细胞中 NADH 绝对浓度的图形方法。该方法需要测量已知浓度的 NADH 溶液。荧光衰减的相量表示解释了图像中逐个像素的游离形式和结合形式 NADH 的量子产率差异。在向相量图中的每个像素添加给定量的未调制光后，获得细胞中每个像素中的 NADH 浓度，这会导致相量向原点移动，移动量取决于像素处的强度和像素处的荧光寿命。通过将图像每个像素处获得的偏移与校准溶液的偏移进行比较来获得 NADH 的绝对浓度。 (C) 2016年美国光学学会
We propose a graphical method using the phasor representation of the fluorescence decay to derive the absolute concentration of NADH in cells. The method requires the measurement of a solution of NADH at a known concentration. The phasor representation of the fluorescence decay accounts for the differences in quantum yield of the free and bound form of NADH, pixel by pixel of an image. The concentration of NADH in every pixel in a cell is obtained after adding to each pixel in the phasor plot a given amount of unmodulated light which causes a shift of the phasor towards the origin by an amount that depends on the intensity at the pixel and the fluorescence lifetime at the pixel. The absolute concentration of NADH is obtained by comparison of the shift obtained at each pixel of an image with the shift of the calibrated solution. (C) 2016 Optical Society of America