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In this work, a targeted phosphorescent nanoprobe is prepared based on a pyrene\/benzophenone (PyBP) guest\u2013host system and further functionalized it with folic acid and cyclic Arg\u2010Gly\u2010Asp peptide for enhanced tumor specificity. In oral squamous cell carcinoma (OSCC) xenograft models, this dual\u2010ligand nanoprobe can help achieve a four\u2013fivefold increase in the tumor signal over the background. Notably, the previously unexplored relationship between tumor stiffness and RTP emission is studied. Mechanical and histological analyses confirmed increased stiffness in OSCC tissues. By modulating the rigidity of tumor phantoms and treating the xenografts with collagenase, a direct correlation is revealed between tumor stiffness and phosphorescence intensity. This study suggests the effectiveness of dual targeting in improving tumor visualization using guest\u2013host\u2010doped RTP nanoprobes. The findings demonstrate that the biomechanical features of the tumor microenvironment enhance phosphorescence emission, highlighting tumor stiffness as a natural amplifier for RTP imaging.<\/jats:p>","DOI":"10.1002\/smll.202506938","type":"journal-article","created":{"date-parts":[[2025,8,26]],"date-time":"2025-08-26T08:26:16Z","timestamp":1756196776000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Tumor Stiffness\u2010Enhanced Room Temperature Phosphorescence Imaging Using Dual\u2010Ligand Nanoprobes"],"prefix":"10.1002","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0081-267X","authenticated-orcid":false,"given":"Han","family":"Cheng","sequence":"first","affiliation":[{"name":"College of Stomatology Shanghai Jiao Tong University &amp; National Center for Stomatology &amp; Shanghai Research Institute of Stomatology  Shanghai 200011 P. R. 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