Delineation of tumor margins is a critical and challenging objective during brain cancer surgery. A tumor‐targeting deep‐blue nanoparticle‐based visible contrast agent is described, which, for the first time, offers in vivo tumor‐specific visible color staining. This technology thus enables color‐guided tumor resection in real time, with no need for extra equipment or special lighting conditions. The visual contrast agent consists of polyacrylamide nanoparticles covalently linked to Coomassie Blue molecules (for nonleachable blue color contrast), which are surface‐conjugated with polyethylene glycol and F3 peptides for efficient in vivo circulation and tumor targeting, respectively. Hydrogel nanoparticles containing a high concentration of visible dye by covalent linkage, with PEGylated surface and conjugated tumor targeting moiety, enable visual delineation of brain tumors in vitro and in vivo. This technology enables color‐guided tumor resection in real time, with no need for extra equipment or special lighting conditions
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