Acoustic-spectrum-compensated photoacoustic microscopy.

Photoacoustic microscopy (PAM) can label-free image oxy- and deoxy-hemoglobin (${{\rm HbO}_2}$HbO and Hb) concentrations in vivo, providing useful information for metabolic researches and diagnostic applications. Conventional PAM assumes a linear relationship between the photoacoustic amplitude and the absorption coefficient. However, many factors, including absorber size, laser pulse width, and frequency response of the ultrasound transducer, may affect the measured acoustic spectrum and the shape of the temporal photoacoustic signal. The ultrasound transducer may weigh the blood vessels differently according to their diameters. In addition, the pulse width also affects the photoacoustic signal amplitude. These factors may cause inaccurate measurement of Hb and ${{\rm HbO}_2}$HbO concentrations. To address this issue, we develop an acoustic-spectrum-compensated optical-resolution PAM (OR-PAM) that corrects the nonuniform acoustic spectrum and makes the quantitative results to be independent of the vessel diameter and pulse width. In dual-wavelength OR-PAM, we demonstrate that the acoustic spectrum compensation can improve the accuracy of oxygen saturation imaging by $\sim{15}% $∼15%.