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Among all the metastases, liver metastasis is the most difficult to detect. As most imaging probes have high liver accumulation, it is difficult to distinguish tiny metastases from normal liver tissue with strong background signal. In this study, the design of a novel second near\u2010infrared window (NIR\u2010II) fluorescence probe for precise detection of carcinoma in situ and liver metastases is presented. The probe called Tg\u2010RGD utilizes a commercially available cyanine dye IR\u2010806 as the signaling moiety, a disulfide bond linker as the responsive moiety, an RGD\u2010capped poly(ethylene glycol) (PEG) as the water soluble enhancer, and the tumor targeting moiety. Tg\u2010RGD shows good glutathione (GSH) responsiveness and selectivity, where its NIR\u2010II fluorescence intensity can enhance 50\u2010fold after activation. In vivo study indicates that Tg\u2010RGD shows much better imaging and targeting effects than Tg\u2010PEG with a similar structure but without RGD moiety for both orthotopic breast cancer and osteosarcoma. Most importantly, Tg\u2010RGD can detect tiny liver metastases with high signal\u2010to\u2010background ratio (3.2). Thus, this study reports a high\u2010performance tumor\u2010specific NIR\u2010II fluorescence probe for in situ and tiny metastatic tumor detection, and may further broaden the applications into related tumor lesions.<\/jats:p>","DOI":"10.1002\/smll.202503257","type":"journal-article","created":{"date-parts":[[2025,5,28]],"date-time":"2025-05-28T09:25:35Z","timestamp":1748424335000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Glutathione\u2010Responsive Near\u2010Infrared\u2010II Fluorescence Probe for Early and Accurate Detection of In Situ and Metastatic Tumors"],"prefix":"10.1002","volume":"21","author":[{"given":"Likun","family":"Yin","sequence":"first","affiliation":[{"name":"State Key Laboratory of Flexible Electronics (LoFE) &amp; Institute of Advanced Materials (IAM) Nanjing University of Posts &amp; Telecommunications  9 Wenyuan Road Nanjing 210023 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pu","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Flexible Electronics (LoFE) &amp; 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