Photoacoustic imaging of sub-diffraction objects with spectral contrast

Photoacoustic imaging couples the chemical specificity of optical absorption with the viewing depth of ultrasound. Systems based on linear array transducers have the versatility to be applied in various (pre-) clinical scenarios but face a trade-off between viewing depth and image resolution depending on transducer frequency and aperture. We propose here a method to disentangle, with precision, small, closely spaced targets with optical spectral contrast, without impairing the imaging depth. Photoacoustic data sets were recorded at two different optical wavelengths. We accurately recovered object separation distances (mean error = 2.3% ô 6%) from the phase difference between signals across the array, down to a spacing of 1/20th of the system's beam-formed lateral resolution. The proposed method may enable the translation of super-resolution microscopy to deep tissue imaging.