NAD(P)H Activated Fluorescent Probe for Rapid Intraoperative Pathological Diagnosis and Tumor Histological Grading

Rapid and accurate intraoperative pathological diagnosis (IOPD) is essential for intraoperative decision-making to improve patients' outcomes and avoid reoperations. In this study, using a NAD(P)H-activated fluorescent probe, a multifunctional fluorescent indicator has been developed to selectively identify tumor cells from normal tissue and to achieve cancer grading identification. This rapid response probe, CyQ-1, features unprecedented sensitivity and rapid response toward NADH at low nanomolar levels under physiological conditions. Moreover, this indicator allows both colorimetric and fluorescent NADH detection in HeLa, A549, MDA-MB-231, 4T1, MCF-7, HePG2, HUVEC, and HL-7702 cells. Expanding the use of this indicator to advanced tissue models, its ability to visualize NADH in 120 paraffin-embedded colorectal sections and 20 cases of intraoperative frozen sections of lung cancer was further verified. CyQ-1-based cancer grading identification shows an overall 92.5 and 100% agreement with the "gold standard test" of histologic grading toward paraffin and frozen sections, respectively. The sensitivity and specificity for discriminating poorly, moderately, and well-differentiated tumor sections were all above 90%. In a word, the rapid and accurate NADH detection ability for clinical sections makes this proposed indicator a potential candidate for clinical IOPD quantification and tumor differentiation grade recognition.