Non-invasive and real-time visualisation of metabolic activities in living small organisms such as zebrafish embryo and larvae has not yet been attempted due to profound analytical limitations of existing technologies. Significant progress in the development of physico-optical oxygen sensors using luminescence quenching by molecular oxygen has recently been made. Sensing using such microsensors is, however, still performed in small glass chambers that hold single specimens and thus not amenable for high-throughput data acquisition.
In this work, we present a proof-of-concept approach by using microfluidic Lab-on-a-Chip (LOC) technologies combined with sophisticated optoelectronic sensors. The LOC device is capable of immobilising live zebrafish embryos with continuous flow perfusion, while the sensor uses innovative Fluorescence Ratiometric Imaging (FRIM) technology that can kinetically quantify the temporal patterns of aqueous oxygen gradients at a very fine scale based on signals coming from an optical sensor referred to as a sensor foil. By embedding the sensor foil onto the microfluidic living embryo array system, we demonstrated in situ FRIM on developing zebrafish embryos. Future integration of microfluidic chip-based technologies with FRIM technology represents a noteworthy direction to miniaturise and revolutionise research on metabolism and physiology in vivo.