These high-speed CRS imaging systems have made significant impacts in the field of vibrational imaging and triggered drastic expansion of the related research, including instrumental developments 7, 8, 9, 10, 11 and biological applications 12, 13, particularly live-cell analysis. Most single-cell vibrational imaging techniques exploit Raman scattering, and the state-of-the-art coherent Raman scattering (CRS) microscopes have achieved high-speed imaging at video rates 5, 6. Vibrational imaging such as Raman scattering and mid-infrared (MIR) absorption imaging has attracted attention in life science 1, 2, e.g., in the field of single-cell biology, because its label-free capability can solve the problems associated with fluorescence imaging, such as cell damage or death due to cytotoxicity, difficulty in continuous and quantitative measurements due to photobleaching, and undesired functional modification of the labeled intracellular biomolecules 3, 4. Our high-speed and high-spatial-resolution MIR microscope has great potential to become a new tool for life science, in particular for live-cell analysis. Then, we develop the designed system with a homemade nanosecond MIR optical parametric oscillator and a high full-well-capacity image sensor. We first derive optimal system design by numerically simulating thermal conduction following the photothermal effect. Here, we develop a significantly improved wide-field MIP quantitative phase microscope with two orders-of-magnitude higher signal-to-noise ratio than previous MIP imaging techniques and demonstrate live-cell imaging beyond video rate. However, the maximum measurement rate has been limited to several frames s −1, limiting its range of use. Recently, mid-infrared photothermal (MIP) imaging has proven to be applicable to 2D and 3D single-cell imaging with high spatial resolution inherited from visible light. On the contrary, MIR microscopy has been rarely used for live biological samples in an aqueous environment due to the lack of spatial resolution and the large water absorption background. Advancement in mid-infrared (MIR) technology has led to promising biomedical applications of MIR spectroscopy, such as liquid biopsy or breath diagnosis.
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