High-speed dynamic 3D photoacoustic imaging of sentinel lymph node in a murine model using an ultrasound array

Noninvasive photoacoustic sentinel lymph node (SLN) mapping with high spatial resolution has the potential to improve the false negative rate and eliminate the use of radioactive tracers in SLN identification. In addition, the demonstrated high spatial resolution may enable physicians to replace SLN biopsy with fine needle aspiration biopsy, and thus reduce the risk of associated morbidity. The primary goal of this study is to demonstrate the feasibility of high-speed 3D photoacoustic imaging of the uptake and clearance dynamics of Evans blue dye in SLNs. The photoacoustic imaging system was developed with a 30 MHz ultrasound array and a kHz repetition rate laser system. It acquires one 3D photoacoustic image of 166 B-scan frames in 1 s, with axial, lateral, and elevational resolutions of 25, 70, and 200 microm, respectively. With optic-fiber based light delivery, the entire system is compact and is convenient to use. Upon injection of Evans blue, a blue dye currently used in clinical SLN biopsy, SLNs in mice and rats were accurately and noninvasively mapped in vivo using our imaging system. In our experiments, the SLNs were found to be located at approximately 0.65 mm below the skin surface in mice and approximately 1.2 mm in rats. In some cases, lymph vessels and lymphatic valves were also imaged. The dye dynamics--accumulation and clearance--in SLNs were quantitatively monitored by sequential 3D imaging with temporal resolution of as high as approximately 6 s. The demonstrated capability suggests that high-speed 3D photoacoustic imaging should facilitate the understanding of the dynamics of various dyes in SLNs and potentially help identify SLNs with high accuracy.