Label-free mid-infrared photothermal live-cell imaging beyond video rate

Advancement in mid-infrared (MIR) technology has led to promising biomedical applications of MIR spectroscopy, such as liquid biopsy or breath diagnosis. 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. 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. However, the maximum measurement rate has been limited to several frames s(-1), limiting its range of use. 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. We first derive optimal system design by numerically simulating thermal conduction following the photothermal effect. Then, we develop the designed system with a homemade nanosecond MIR optical parametric oscillator and a high full-well-capacity image sensor. 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.

Automated summary

Advancement in mid-infrared (MIR) technology has led to promising biomedical applications, but MIR microscopy is limited for live biological samples due to the lack of spatial resolution and water absorption. However, the development of a wide-field MIP quantitative phase microscope with improved signal-to-noise ratio allows for live-cell imaging beyond video rate. This high-speed and high-spatial-resolution MIR microscope has great potential for live-cell analysis and can become a new tool in life science.