Near-infrared double-illumination optical-resolution photoacoustic microscopy

Label-free chemical bond imaging is of great importance in biology and medicine. Photoacoustic imaging at the third near-infrared windows (1600-1870 nm, near-infrared-III) provides a stable molecular vibrational imaging tool for lipid-rich tissue owing to the first overtone transition of the CH bond at 1.7 μm. However, lacking high-energy pulsed laser sources at 1.7 μm and the strong water absorption significantly limit the signal-to-noise ratio of the lipid imaging, especially for thin lipid tissues. To circumvent this barrier, we develop near-infrared-III double-illumination optical-resolution photoacoustic microscopy (DIOR-PAM) for improving the sensitivity of label-free lipid imaging. Using the same laser, DIOR-PAM can enhance the sensitivity by nearly 100%, which we prove in the Monte Carlo simulation. We experimentally demonstrated 50% ~ 100% sensitivity enhancements on nonbiological and biological lipid-rich samples.