Fluorescence-Detected Mid-Infrared Photothermal Microscopy

We demonstrate instrumentation and methods to enable fluorescence-detected photothermal infrared (F-PTIR) microscopy and then demonstrate the utility of F-PTIR to characterize the composition within phase-separated domains of model amorphous solid dispersions (ASDs) induced by water sorption. In F-PTIR, temperature-dependent changes in fluorescence quantum efficiency are shown to sensitively report on highly localized absorption of mid-infrared radiation. The spatial resolution with which infrared spectroscopy can be performed is dictated by fluorescence microscopy, rather than the infrared wavelength. Intrinsic ultraviolet autofluorescence of tryptophan and protein microparticles enabled label-free F-PTIR microscopy. Following proof of concept F-PTIR demonstration on model systems of polyethylene glycol (PEG) and silica gel, F-PTIR enabled the characterization of chemical composition within inhomogeneous ritonavir/polyvinylpyrrolidone-vinyl acetate (PVPVA) amorphous dispersions. Phase separation is implicated in the observation of critical behaviors in ASD dissolution kinetics, with the results of F-PTIR supporting the formation of phase-separated drug-rich domains upon water sorption in spin-cast films.

Automated summary

This study demonstrates the utility of fluorescence-detected photothermal infrared (F-PTIR) microscopy in characterizing phase-separated domains within model amorphous solid dispersions (ASDs) induced by water sorption. The temperature-dependent changes in fluorescence quantum efficiency sensitively report on localized absorption of mid-infrared radiation. The spatial resolution of infrared spectroscopy in F-PTIR is determined by fluorescence microscopy, enabling high-resolution analysis of chemical composition.