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The objective of this work is to showcase the ortho-positronium lifetime as a probe for soft-tissue characterization. We employed positron annihilation lifetime spectroscopy to experimentally measure the three components of the positron annihilation lifetime—para-positronium (p-Ps), positron, and ortho-positronium (o-Ps)—for three types of porcine, non-fixated soft tissues ex vivo: adipose, hepatic, and muscle. Then, we benchmarked our measurements with X-ray phase-contrast imaging, which is the current state-of-the-art for soft-tissue analysis. We found that the o-Ps lifetime in adipose tissues (2.54 ± 0.12 ns) was approximately 20% longer than in hepatic (2.04 ± 0.09 ns) and muscle (2.03 ± 0.12 ns) tissues. In addition, the separation between the measurements for adipose tissue and the other tissues was better from o-Ps lifetime measurement than from X-ray phase-contrast imaging. This experimental study proved that the o-Ps lifetime is a viable non-invasive probe for characterizing and classifying the different soft tissues. Specifically, o-Ps lifetime as a soft-tissue characterization probe had a strong sensitivity to the lipid content that can be potentially implemented in commercial positron emission tomography scanners that feature list-mode data acquisition. [ABSTRACT FROM AUTHOR]