摘要/Abstract

生物体内活性小分子（生物小分子），不仅数量多，涉及到无机小分子，如SO₂，H₂S，NO与CO等，而更多的是有机小分子，如单煻、低聚糖、激素、辅酶、甘油、刺激因子、调节因子和维他命等；而且在病理、生理等方面有着至关重要的功能，发挥着举足轻重的作用，直接影响身体健康与疾病的发生.因此，对生物体内小分子的实时观测和监控是十分必要的，而双光子荧光探针正是实现这一目标的必选锐器.双光子荧光探针的定点激发、高横向与纵向分辨率、无光漂白性、无光致毒性和组织深度成像等诸多优点而彰显其无与伦比的优越性，可以很好地对细胞或组织内的生物小分子进行实时动态三维观测和监控.综述了近5年来相继开发的CO、单糖（葡萄糖、β-半乳糖苷酶）、SO₂、硫化氢、NO、过氧（硫）化物、硫醇/硫酚、¹O₂、甲醛、HNO、HClO、O₂^·-和ONOO^-等双光子荧光探针，系统全面地剖析了这些双光子荧光探针的传感机制，并展望了生物小分子显微用双光子荧光探针的发展与愿景.
关键词: 生物小分子, 荧光探针, 传感机制, 荧光团, 受体, 双光子显微
Small molecules (biological small molecules) in vivo are not only in a large number, involving inorganic small molecules, such as SO₂, H₂S, NO and CO, and more organic small molecules, such as monosaccharides, oligosaccharides, hormones, coenzymes, glycerol, stimulating factors, regulatory factors, Vitamins, etc., moreover, play an important role in pathology, physiology, and so on. Therefore, it is necessary to observe and monitor small molecules in vivo in real time, and the two-photon fluorescence probe is a necessary tool to achieve this goal. The advantages of two-photon fluorescence probe, such as fixed target (very small dot) excitation, high horizontal and vertical resolution, no photobleaching, no phototoxicity and deep imaging in tissues, and so on, demonstrate its unparalleled superiority. It can be used for dynamic 3D observation and monitoring of biological small molecules in cells or tissues. In this paper, CO, monosaccharide (glucose, β-galactosidase), SO₂, H₂S, NO, peroxy (sulfur) compounds, mercaptan/thiophenol, ¹O₂, formaldehyde, HNO, HclO, O₂^·- and ONOO^- two-photon fluorescence probes which have been developed in recent 5 years were reviewed. The sensing mechanisms of these two-photon fluorescence probes were systematically analyzed, and the development and prospect of two-photon fluorescence probes for small biomolecules are prospected.
Key words: biological small molecule, fluorescent probe, sensing mechanism, fluorophore, receptor, two-photon bioimaging


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