Polymer microparticles containing an immobilized pH-sensitive dye are used for determination of pH inside microfluidic segments. The particles possess a hydrophilic surface in order to get a homogenous distribution inside the aqueous phase of microfluidic segments. The dye is coupled to the polymer matrix by a covalent bond. The pH can be determined by the read-out of fluorescence intensity. In contrast to dissolved indicator dyes, the chemical interference of the sensor particles with the surrounding solution is negligible. So, the particle-based sensing can easily be applied to the determination of pH changes during the cultivation of cells inside the microfluidic segments. The typical change of pH during cell cultivation can be used for monitoring the kinetic of cell cultivation inside single volumes in the nanoliter range, so that information about the metabolic activity of the organism can be obtained. An LED-based miniaturized flow-through fluorometer was developed to determine the fluorescence directly inside microtubes of an inner diameter of 0.5 mm. It allows measurement frequencies up to 60 Hz and is suited for characterization of fast moving large sequences of microfluidic segments.