Synchrotron radiation infrared microspectroscopy of single living cells in microfluidic devices: Advantages, disadvantages and future perspectives

The possibility to fully exploit the diagnostic capabilities of SR-IRMS for studying single living cells under physiological conditions is limited by several constrains. First of all, the technology for manufacturing materials transparent to both IR and visible light is quite immature, limiting the design of fluidic devices to simple demountable liquid cells. In addition, the water spectral features become prominent in the Mid IR, hiding several cellular bands and therefore limiting the diagnostic capabilities of SR-IRMS. The overcoming of the so called "water absorption barrier" requires the improvement of the protocols for the compensation of buffer spectral contributions, a goal that can be achieved also advancing the quality of IR-suitable fluidic devices. In this paper, the technical solutions employed for microfabricating completely sealed IR-visible transparent fluidic devices for living cell analysis will be presented. Several examples of the results obtained in the study of living U937 monocytes subjected to different stimuli will be selected for highlighting both the advantages and the disadvantages offered by our approach for cellular biology.