Real-time functional optical-resolution photoacoustic microscopy using high-speed alternating illumination at 532 and 1064 nm

Optical-resolution photoacoustic microscopy (OR-PAM), which has been widely used and studied as a noninvasive and in vivo imaging technique, can yield high-resolution and absorption contrast images. Recently, metallic nanoparticles and dyes, such as gold nanoparticles, methylene blue, and indocyanine green, have been used as contrast agents of OR-PAM. This study demonstrates real-time functional OR-PAM images with high-speed alternating illumination at 2 wavelengths. To generate 2 wavelengths, second harmonic generation at 532 nm with an LBO crystal and a pump wavelength of 1064 nm is applied at a pulse repetition rate of 300 kHz. For alternating illumination, an electro-optical modulator is used as an optical switch. Therefore, the A-line rate for the functional image is 150 kHz, which is half of the laser repetition rate. To enable fast signal processing and real-time displays, parallel signal processing using a graphics processing unit (GPU) is performed. OR-PAM images of the distribution of blood vessels and gold nanorods in a BALB/c-nude mouse's ear can be simultaneously obtained with 500 x 500 pixels and real-time display at 0.49 fps.