Ultra long-lifetime and high-sensitive fluorescent measurement using difference compensation method for single cell analysis

In this paper, we proposed long-lifetime and high-sensitive measurement using fluorescent intensity by compensation of photo-bleaching with the difference compensation for cell measurement. Photo-bleaching of fluorescence is one of the most crucial factors to cause measurement error in physiological measurement. In our approach, difference of fluorescent intensity between imaging time is compensated using single exponential function to make the difference amount uniform in each imaging time. We evaluated the dependencies of measurable time and the sensitivity by fluorescent lifetime and the decay coefficient of fluorescent lifetime using numerical simulation. From these results, the proposed method has much higher sensitivity and longer measurable time against conventional compensation method of photo-bleaching. By using the proposed method, we demonstrate the photosynthesis activity of single Synechocystis sp. PCC 6803 in the micro chamber with gas barrier layer.