The imaging speed of the current mid-infrared photothermal (MIP) microscope is limited to tens of seconds per frame due to the long pixel dwell time and slow sample scanning process, which is insufficient for capturing dynamics inside living systems. In this work, we developed a video-rate MIP microscope by employing a lock-in free demodulation scheme to resolve single IR pulse induced contrast. We further developed a synchronous pump-probe Galvo scanning for reaching a line rate over 2.5 kHz. The system is capable of resolving chemical dynamics in living cells in a uniform imaging field of view over 300 μm.
Optical coherence tomography (OCT) has been a powerful 3D optical imaging tool in the last decade while it lacks molecular information. In this work, we integrate the mid-infrared photothermal microscopy with the OCT approach to demonstrate a bond-selective full-field optical coherence tomography (BS-FF-OCT), in which a pulsed mid-infrared laser is used to modulate the full-field OCT signal through the photothermal effect. This method achieves label-free volumetric infrared spectroscopic imaging at 1-μm isotropic resolution, demonstrated by a variety of samples, including 1 μm PMMA beads embedded in agarose gel, polypropylene fiber mattress, myelinated nerve bundle in mouse brain tissue, Caenorhabditis elegans, and cancer cell spheroids.
Volumetric chemical imaging is highly desired for investigating biochemical processes at the sub-cellular level. Here, we report bond-selective intensity diffraction tomography (BS-IDT) based on 3D quantitative phase detection of the mid-infrared photothermal effect. BS-IDT demonstrates volumetric chemical imaging with incoherent diffraction-limited resolution and a high speed up to ~6 Hz per volume. The mid-IR spectrum extracted from BS-IDT shows high fidelity compared with ground truth measured by an FTIR spectrometer. The 3D chemical imaging results from cancer cells and Caenorhabditis elegans validate BS-IDT’s superior performance.
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