Fluorescence-sensing methods

Novel approaches to sensor design, based on the use of an internal standard with appropriate spectral properties, provide new possibilities for designing simple devices for fluorescence sensing. Detection of combined emission from the reference and an analyte-sensitive fluorophore has been achieved in numerous measurements in cuvettes, high-throughput formats. These methods have been used with a long-lifetime reference to measure pH, O₂, pCO₂, glucose, and calcium by means of modulation-sensing methods as well as by the use of oriented films as the reference for polarization sensing of glucose, pH, oxygen, and lactate. Polarization sensing has also been developed with visual detection to measure the concentration of rhodamine B and pH. Modulation and polarization sensing was found to be effective in highly scattering media such as Intralipid or tissue. The applicability of these technologies to transdermal diagnostics depends on the availability of red fluorophores that can be used in vivo. One dye that could possibly be used is indocyanine green (IcG), which absorbs and emits at wavelengths above 700 nm. Furthermore, IcG has already been approved for use in humans for monitoring burn severity and it has been detected through the skin. It appears likely that modern optics and electronic technology will allow the development of practical devices for biomedical use as shown in Scheme 1.