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Purpose: The visibility of biopsy needles in contrast-specific imaging mode can be improved by priming them with an ultrasound contrast agent (previously demonstrated in a phantom model/ex vivo). The purpose of this study was to validate this priming method in a porcine in vivo model. Materials and Methods: Using a small syringe, full-core biopsy needles were primed with sulfur hexafluoride, an ultrasound contrast agent, with non-primed needles serving as controls (n = 30 + 30). Liver punctures were performed in a porcine model following intravenous administration of the same ultrasound contrast agent. Needle visibility, both in their entirety and at the tips, was evaluated in split-screen mode using contrast-specific imaging and B-mode (low mechanical index). The assessment included quantitative analysis, calculating the contrast-to-noise ratio, and qualitative evaluation through structured grading by three radiologists. Results: After needle priming, the contrast-to-noise ratio was superior for the needle in its entirety in contrast-specific imaging mode (p < 0.001) and slightly inferior in B-mode (p = 0.008). No differences were observed for the needle tips in either imaging mode. Qualitatively, the needle visibility was deemed clinically superior after needle priming throughout in contrast-specific imaging mode (p < 0.001), whereas no clinically relevant differences in B-mode for either the needle in its entirety (p = 0.11) or the needle tip (p = 1) were observed. Conclusion: In this in vivo porcine liver model experiment, priming biopsy needles with ultrasound contrast agent improved needle visibility in contrast-specific imaging mode but slightly reduced it in B-mode. These findings support the method's use for biopsies requiring target visualization in contrast-specific imaging mode. No level of evidence. [ABSTRACT FROM AUTHOR]