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Percutaneous radiofrequency ablation (RFA) is gaining importance as a locoregional treatment for tumors in several organs including the liver, lung, kidney and bone. In RFA, the tumor is eradicated with the direct application of heat using alternating current through a needle electrode positioned under imaging guidance. Various imaging methods are used in the RFA ablation procedure but these have drawbacks. In this work, we introduce photoacoustic (PA) imaging as a new method with potential to visualize the targeting of RFA needle into a region of interest and to report on the extent of ablation achieved. We demonstrate the proof-of-concept in using PA imaging together with ultrasound imaging on ex vivo biological samples in the laboratory simulating relevant clinical scenarios in RFA. These include guidance of the RFA needle to target tissue, mapping of simulated blood vessels during needle insertion and …