Ratiometric pH measurements using LysoSensor DND-192

A method for the ratiometric pH sensing using LysoSensor DND-192 is presented in this paper. It works in the physiological pH range. It is based on the use of two fluorophores which differ significantly in their lifetimes. As the discrimination of their emissions is performed through two different frequencies, this method can allow significant overlap of the emission spectra. A simple long-pass filter, or a combination of long- and short-pass filters, was used instead of narrow-bandpass devices. Importantly, the measurements were carried out under strong ambient light. The method could be used in a wide variety of applications, such as intracellular measurements, microscopy, bioprocess monitoring, etc.