Opt Lett. 2024 Jul 15;49(14):4054-4057. doi: 10.1364/OL.529284.
Two-photon autofluorescence (TPAF) imaging is able to offer precise cellular metabolic information with high spatiotemporal resolution, making it a promising biopsy tool. The technique is greatly hampered by the complexity of either the optical system or data processing. Here, the excitation wavelength was optimized to simultaneously excite both flavin adenine dinucleotide and nicotinamide adenine dinucleotide and eliminate the unexpected TPAF. The optical redox ratio (ORR) images were robustly achieved without additional calibration under the optimized single-wavelength excitation. The in vitro, ex vivo, and in vivo biopsy by the TPAF method were systematically studied and compared using hepato-cellular carcinoma and metastasis as examples. It was demonstrated that the proposed TPAF method simplified the optical system, improved the robustness of ORR, and enabled early-stage cancer diagnosis, showing distinguished advantages as compared with previous methods.
双光子自发荧光（TPAF）成像能够提供具有高时空分辨率的精确细胞代谢信息，是一种很有前途的活检工具。该技术受到光学系统或数据处理复杂性的极大限制。在这里，优化了激发波长，以同时激发黄素腺嘌呤二核苷酸和烟酰胺腺嘌呤二核苷酸，并消除意外的 TPAF。在优化的单波长激发下，无需额外的校准，即可稳健地获得光学氧化还原比（ORR）图像。使用肝癌和转移作为示例，系统地研究并比较了 TPAF 方法的体外、离体和体内活检。结果表明，所提出的 TPAF 方法简化了光学系统，提高了 ORR 的稳健性，并能够进行早期癌症诊断，与以前的方法相比具有明显的优势。
