Free-electron laser-based extended wide-field mid-infrared photothermal imaging for biomedical and microplastic analysis

Wide-field mid-infrared photothermal (MIP) imaging offers rapid labelfree chemical contrast for biomedical and polymer analysis. However, its field of view (FOV) is limited by the pulse intensity of conventional infrared lasers. Here, we present a wide-field MIP microscope that uses a high-power free-electron laser (FEL) rather than a quantum cascade laser (QCL) as the pump source to achieve a substantially larger FOV. Both implementations use counter-propagating beam paths with a 450 nm LED as the probe source and a CMOS camera that records images using a virtual lock-in detection scheme. QCL nanojoule pulse energies enables FOV of around 45 micrometers for widefield MIP imaging with a sub-micrometer resolution for polystyrene beads, Mycobacterium tuberculosis infected fixed tissues, and laryngeal cancer cryosections. IR spectra in the range of 1000-1800 wavenumbers can be reconstructed by tuning the QCL. FEL pulse energies of up to microjoules expand the FOV by a factor of nearly 20 as demonstrated by wide-field MIP imaging of polystyrene beads, single cells, and murine brain tissue. We discuss current challenges and further improvements to implement high-power IR lasers for wide-field MIP imaging with even larger FOVs in the context of biomedical research and diagnostics.