Development of novel micro-spectroscopy methods

To study complex uptake processes in situ or in vivo, it is necessary to access multiple information regimes in a correlated manner. For biological samples one would like to have state-of-the-art single molecule fluorescence imaging methods (e.g. multiple excitation wavelength, two-photon excitation and fluorescence live time imaging (FLIM)). For characterizing solid particles on the other hand, it is preferable to employ label-free methods to avoid influencing the structural integrety of the samples. Non-linear imaging techniques, such as second harmonic generation (SHG), four-wave mixing (FWM), or the chemically sensitive coherent Anti-Stokes Raman Scattering (CARS), enable label-free studies. Uptake of particles in living cells will result in complex chemical interactions, best interrogated with spectroscopy. A combination of all these methods is crucial to entangle the complex mechanisms and kinetics underlying uptake processes. We therefore developed a Multimodal Optical Spectroscopy And Imaging Correlation (MOSAIC) analysis platform based on non-linear confocal micro-/spectroscopy. It combines the afore mentioned methods and has already shown that a correlative approach is crucial e.g. in studying absorption processes in MOF crystals. We are currently adding stimulated Raman scattering (SRS) and tracking methods to our toolbox.

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