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Therefore, the single molecule for simultaneous therapy and feedback has become a promising tool. Herein, we develop an in situ self\u2010assembled probe (HPO\u2010CYFF\u2010TPP) for cascade\u2010instructed therapy and feedback imaging. Under the dephosphorylation of alkaline phosphatase, HPO\u2010CYFF\u2010TPP converts to HO\u2010CYFF\u2010TPP and self\u2010assembles into nanoparticles (NPs) with quenched fluorescent (FL) and opened photoacoustic (PA) signal. After laser irradiation, NPs can produce abundant singlet oxygen for photodynamic therapy (PDT) of tumor cells, which leading to the increase of intracellular viscosity. At high viscosity, NPs opens FL and further enhances PA signal. HPO\u2010CY\u2010TPP without self\u2010assembly group is a control probe. HPO\u2010CYFF\u2010TPP or HPO\u2010CY\u2010TPP exhibits effective PDT against tumor cells and high tumor inhibition in tumor\u2010bearing mice, while HPO\u2010CYFF\u2010TPP is more effective than HPO\u2010CY\u2010TPP due to the formation of NPs in situ. Importantly, HPO\u2010CYFF\u2010TPP or HPO\u2010CY\u2010TPP can monitor the viscosity during PDT on tumor cells and tumor\u2010bearing mice for treatment feedback, while HPO\u2010CYFF\u2010TPP is more accurate than HPO\u2010CY\u2010TPP with higher FL\/PA sensitivity. We envision that this self\u2010assembled probe will be a powerful tool for tumor therapy and real\u2010time treatment feedback imaging in the future.<\/jats:p>","DOI":"10.1002\/smll.75259","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T09:20:01Z","timestamp":1786699201000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["In Situ Self\u2010Assembled Nanoparticles for Cascade\u2010Instructed Photodynamic Therapy and Real\u2010Time Treatment Feedback Imaging"],"prefix":"10.1002","volume":"22","author":[{"given":"Taoli","family":"Sun","sequence":"first","affiliation":[{"name":"Institutes of Physical Science and Information Technology Anhui University  Hefei Anhui China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenbin","family":"Liu","sequence":"additional","affiliation":[{"name":"Institutes of Physical Science and Information Technology Anhui University  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