In vivo deconvolution acoustic-resolution photoacoustic microscopy in three dimensions

Acoustic-resolution photoacoustic microscopy (ARPAM) provides a spatial resolution on the order of tens of micrometers, and is becoming an essential tool for imaging fine structures, such as the subcutaneous microvasculature. High lateral resolution of ARPAM is achieved using high numerical aperture (NA) of acoustic transducer; however, the depth of focus and working distance will be deteriorated correspondingly, thus sacrificing the imaging range and accessible depth. The axial resolution of ARPAM is limited by the transducer's bandwidth. In this work, we develop deconvolution ARPAM (D-ARPAM) in three dimensions that can improve the lateral resolution by 1.8 and 3.7 times and the axial resolution by 1.7 and 2.7 times, depending on the adopted criteria, using a 20-MHz focused transducer without physically increasing its NA and bandwidth. The resolution enhancement in three dimensions by D-ARPAM is also demonstrated by in vivo imaging of the microvasculature of a chick embryo. The proposed D-ARPAM has potential for biomedical imaging that simultaneously requires high spatial resolution, extended imaging range, and long accessible depth.