To assure complete
tumor removal, frozen section analysis is the most common procedure for intraoperative pathological assessment of resected
tumor margins. However, during one operation, multiple biopsies may be sent for examination, but only few of them are made into cryosections because of the complex preparation protocols and time-consuming pathological analysis, which potentially increases the risk of overlooking
tumor involvement. Here, we propose a fluorescence-based pre-screening strategy that allows high-throughput, convenient, and fast gross assessment of resected
tumor margins. A dual-activatable cationic fluorescent molecular rotor was developed to specifically illuminate live
tumor cells' cytoplasm by emitting two different fluorescence signals in response to elevations in
hypoxia-induced
nitroreductase (a
biochemical marker) and cytoplasmic viscosity (a biophysical marker), two characteristics of
cancer cells. The ability of the fluorescent molecular rotor in detecting
tumor cells was evaluated in mouse and human specimens of multiple tissues by comparing with
hematoxylin and
eosin staining. Importantly, the fluorescent molecular rotor achieved 100 % specificity in discriminating lung and
liver cancers from normal tissue, allowing pre-screening of the
tumor-free
surgical margins and promoting clinical decision. Altogether, this type of fluorescent molecular rotor and the proposed strategy may serve as a new option to facilitate intraoperative assessment of resected
tumor margins.