State Key Laboratory of Fine Chemicals, School of Chemistry, Dalian University of Technology, Dalian 116024, PR China.
Talanta. 2011 May 15;84(3):971-6. doi: 10.1016/j.talanta.2011.02.051. Epub 2011 Mar 4.
Time-resolved (or time-gated) luminescence detection technique using lanthanide chelates as luminescent probes is a widely used and highly sensitive method for the biological applications. The developments of various functional lanthanide probes that can selectively recognize the biological targets are the essential objective of the technique. In this work, a unique Tb(3+) chelate-based luminescent probe, N,N,N(1),N(1)-[2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-(p-aminophenoxy)methylene-pyridine] tetrakis(acetate)-Tb(3+)(BMPTA-Tb(3+)), has been designed and synthesized for highly selective and sensitive time-resolved luminescence detection of one highly reactive oxygen species (ROS), hydroxyl radical (OH). The probe is almost non-luminescent, and can selectively react with hydroxyl radical to afford a highly luminescent Tb(3+) chelate, N,N,N(1),N(1)-[2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-hydroxymethyl-pyridine] tetrakis(acetate)-Tb(3+) (BHTA-Tb(3+)), accompanied by a 49-fold increase in luminescence quantum yield with a long luminescence lifetime (2.76 ms). The luminescence response of the probe to hydroxyl radical is highly selective and insensitive to pH in the physiological pH range. For loading the probe into the living cells, the acetoxymethyl ester of BMPTA-Tb(3+) was synthesized and used for the HeLa cell loading. Based on this probe, a background-free time-resolved luminescence imaging method for detecting hydroxyl radical in living cells was successfully established.
基于镧系配合物作为荧光探针的时间分辨（或时间门控）荧光检测技术是一种广泛应用且高度灵敏的生物应用方法。开发能够选择性识别生物靶标的各种功能镧系探针是该技术的重要目标。在这项工作中，设计并合成了一种独特的基于 Tb(3+)配合物的荧光探针，N,N,N(1),N(1)-[2,6-双(3'-氨基甲基-1'-吡唑基)-4-(对氨基苯氧基)亚甲基吡啶]四乙酸-Tb(3+)(BMPTA-Tb(3+))，用于高度选择性和灵敏的时间分辨荧光检测一种高反应性氧物种（ROS），羟基自由基（OH）。探针几乎没有荧光，并且可以选择性地与羟基自由基反应，生成高荧光 Tb(3+)配合物，N,N,N(1),N(1)-[2,6-双(3'-氨基甲基-1'-吡唑基)-4-羟甲基吡啶]四乙酸-Tb(3+)(BHTA-Tb(3+))，同时荧光量子产率增加 49 倍，荧光寿命延长（2.76 ms）。探针对羟基自由基的荧光响应具有高度选择性，并且在生理 pH 范围内对 pH 不敏感。为了将探针载入活细胞中，合成了 BMPTA-Tb(3+)的乙氧甲酯，并用于 HeLa 细胞的载入。基于该探针，成功建立了用于检测活细胞中羟基自由基的无背景时间分辨荧光成像方法。
