Quantitative phase imaging with molecular vibrational sensitivity

Quantitative phase imaging (QPI) quantifies the sample-specific optical-phase-delay enabling objective studies of optically transparent specimens such as biological samples but lacks chemical sensitivity, limiting its application to a morphology-based diagnosis. We present wide-field molecular vibrational (MV) microscopy realized in the framework of QPI utilizing a mid-infrared (MIR) photothermal effect. Our technique provides MIR spectroscopic performance comparable to that of a conventional infrared spectrometer in the molecular fingerprint region of 1450−1640 cm−1 and realizes wide-field molecular imaging of a silica-polystyrene bead mixture over a 100 μm×100 μm area at 1 frame per second with the spatial resolution of 430 nm and 2–3 orders of magnitude lower fluence of ∼10 pJ/μm2 compared to other high-speed label-free molecular imaging methods, reducing photodamages to the sample. With a high-energy MIR pulse source, our technique could enable high-speed, label-free, simultaneous, and in situ acquisition of quantitative morphology and MV contrast, providing new insights for studies of optically transparent complex dynamics.