In Vivo Reflection-Mode Photoacoustic Microscopy Enhanced by Plasmonic Sensing with an Acoustic Cavity

Relying on high-sensitivity refractive index sensing and a highly constrained evanescent field of surface plasmon resonance (SPR), broadband photoacoustic (PA) pressure transients were measured using an SPR sensor instead of routinely used piezoelectric ultrasonic transducers. An acoustic cavity made from stainless steel and having a designed ellipsoidal inner surface redirected laser-induced PA waves from the PA excitation spot to the SPR sensor. By incorporating the SPR sensor with the acoustic cavity, we developed optical-resolution photoacoustic microscopy (OR-PAM) with multiple advantages, including reflection-mode signal capture, improved PA detection sensitivity, increased PA spectral bandwidth as broad as ∼98 MHz, and micrometer-scale lateral resolution. This allowed label-free volumetric PA imaging of vasculature in not only the thin ear but also the thick forelimb of living mice. With these combined advantages, our OR-PAM system potentially offers more opportunities for biomedical investigation, for example, when studying microcirculations in the eye and cortex.