Mid-infrared metabolic imaging with vibrational probes

Understanding metabolism is indispensable to unraveling the mechanistic basis of many physiological and pathological processes. However, in situ metabolic imaging tools are still lacking. Herein we introduce a framework for mid-infrared (MIR) metabolic imaging by coupling the emerging high-information-throughput MIR microscopy with specifically designed IR-active vibrational probes. Three categories of small vibrational tags including azide bond, (13)C-edited carbonyl bond and deuterium-labeled probes are presented to interrogate various metabolic activities in cells, small organisms and mice. Two MIR imaging platforms are implemented including broadband Fourier transform infrared (FTIR) microscopy and discrete frequency infrared (DFIR) microscopy with a newly incorporated spectral region (2000–2300 cm(−1)). Our technique is uniquely suited for metabolic imaging with high-information-throughput. In particular, we performed single-cell metabolic profiling including heterogeneity characterization, and large-area metabolic imaging at tissue/organ level with rich spectral information.