This “pioneer” configuration recently developed and validated at PICT (Platform for Imaging Cells and Tissues, DDUV Institute, UCLouvain-Brussels) is very useful to make Fluorescence Lifetime Imaging Microscopy (FLIM).
FLIM is an imaging technique based on the differences in the decay rate of the photon emission of a fluorophore and which uses the fluorescence lifetime of the fluorophore rather than its intensity.
Why and when was the Airyscan microscope installed at PICT?
The Airyscan microscopy is a new generation confocal microscopy able to reach higher resolution, higher speed and higher sensitivity than classical confocal microscopy. The last microscope acquired by the PICT platform, i.e. the Zeiss LSM980, is based on this methodology. It was installed in 2020 and is now fully operational.
It is adapted for the analysis of a wide range of biological samples, from isolated proteins, small vesicles and single cells to tissues and living mice. It gives excellent results in colocalization studies, spectral analysis and visualization of thick samples.
Why coupling the Airyscan with multiphoton microscopy?
Besides the above applications, the combination of the Airyscan methodology with a laser multiphoton makes this new microscope a unique instrument very useful to answer a number of biological questions requiring FLIM.
This technique allows to study the microenvironment of fluorochromes without the disturbing effects of photon scattering, photobleaching or local concentration effects of the fluorescent probe.
FLIM has been developed and validated at PICT to evaluate the membrane fluidity of bacteria (Mozaheb et al, mSphere 2022). Other FLIM-related applications are in the pipeline, including evaluation of human red blood cell membrane fluidity upon senescence and fluorescent molecule interaction by the Fluorescence Resonance Energy Transfer (FRET)-FLIM method.
