Optical-resolution photoacoustic microscopy (OR-PAM), has been widely used and studied as noninvasive and in-vivo imaging technique, can achieve a high resolution and high contrast image. OR-PAM is combined with optical absorption contrast and detection of acoustic wave generated by thermal expansion. Recently, nanoparticles and dyes have been used as contrast agents of OR-PAM. To obtain functional OR-PAM image such as a distribution image of blood vessels and nanoparticles, a tunable dye laser or optical parametric oscillator (OPO) should be needed at more two wavelength. However, because these lasers have a low pulse repetition rate (10 Hz ~ 10 kHz), a functional OR-PAM image with real-time display has been limited.
In our previous study, we demonstrated high-speed OR-PAM using an Ytterbium fiber laser and a graphics processing unit (GPU) technique at 300 kHz-pulse repetition rates. Although this Ytterbium fiber laser has a high pulse repetition rate, it is not comfortable for functional imaging owing to lasing at only single wavelength. Therefore, in this study, we used a high-speed interlaced illumination method at 532 nm and 1064 nm for real-time display functional OR-PAM. For high-speed interlaced illumination of two wavelength, we applied second harmonic generation effect and a high-speed optical switching using an electro-optic modulator. Therefore, we could obtain maximum amplitude projection (MAP) images about distributions of blood vessels and nanoparticles, simultaneously, with 500 x 500 pixels and a real-time display of approximately 0.5 fps.