Mid-infrared (mid-IR) spectroscopy and optoacoustic/optothermal (OA/OT) imaging are perfectly complementary technologies to each other. Vibrational molecular excitations by mid-IR absorption are utterly de-excited in the form of heat while efficient OA/OT signal generation primarily depends on heat deposition. This synergy allows overcoming the (otherwise) persistent limitations of traditional mid-IR spectroscopy and imaging in live-cell/fresh-tissue applications— i.e., sample opacity due to water absorption. Combination of mid-IR excitation and OA/OT detection has resulted in new tools for label-free live-cell, tissues, and in vivo metabolic research. Here we discuss basic principles on mid-IR detection for spectroscopy and imaging as well as the most recent developments on mid-IR OA and OT microscopy that overcome the limitations of conventional vibrational spectroscopy for biosensing and label-free metabolic microscopy.
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