Fluorescence anisotropy (FA) has been widely applied for detecting and monitoring special targets in life sciences. However, matrix autofluorescence restricted its further application in complex biological samples. Herein, we report a near-infrared-II (NIR-II) FA strategy for detecting adenosine triphosphate (ATP) in human serum samples and breast cancer cell lysate, which employed NIR-II fluorescent Ag₂Se quantum dots (QDs) as tags to reduce matrix autofluorescence effect and applied graphene oxide (GO) to enhance fluorescence anisotropy signals. In the presence of ATP, the recognition between NIR-II Ag₂Se QDs labeled aptamer (QD-pDNA) and ATP led to the release of QD-pDNA from GO, resulting in the obvious decrease of FA values. ATP could be quantitatively detected in concentrations ranged from 3 nM to 2500 nM, with a detection limit down to 1.01 nM. This study showed that the developed NIR-II FA strategy could be applied for detecting targets in complex biological samples and had great potential for monitoring interactions between biomolecules in biomedical research.

荧光各向异性（FA）已被广泛用于检测和监视生命科学中的特殊目标。然而，基质自发荧光限制了其在复杂生物样品中的进一步应用。在这里，我们报告了一种近红外II（NIR-II）FA策略，用于检测人血清样品和乳腺癌细胞裂解物中的三磷酸腺苷（ATP），该策略采用了NIR-II荧光Ag ₂ Se量子点（QD）作为标签降低基质自发荧光效应，并应用氧化石墨烯（GO）增强荧光各向异性信号。在ATP的存在下，NIR-II Ag ₂之间的识别Se标记的适体（QD-pDNA）和ATP导致了GO中QD-pDNA的释放，导致FA值明显降低。ATP的定量浓度范围为3 nM至2500 nM，检出限低至1.01 nM。这项研究表明，已开发的NIR-II FA策略可用于检测复杂生物样品中的目标，并且在生物医学研究中具有监测生物分子之间相互作用的巨大潜力。