Department of Chemistry, Xinzhou Teachers University, Xinzhou, Shanxi 034000, China.
Analyst. 2017 May 2;142(9):1545-1553. doi: 10.1039/c7an00058h.
A two-photon fluorescent probe FNTR for nitroreductase was synthesized by using 9,9-dimethyl-2-acetyl-fluoren-7-methylamino (1) as a two-photon fluorophore and a p-nitrobenzyl carbamate group as a recognition domain for nitroreductase (NTR). The probe and the fluorophore were tested under one- and two-photon modes respectively. After reacting with nitroreductase, FNTR had a 130-fold fluorescence enhancement at 563 nm in 10 min and the maximal two-photon action cross-section value was detected as 66 GM at 750 nm. The probe showed a high sensitivity with a detection limit as low as 23.67 ng ml, high selectivity, low cytotoxicity and good photostability. In the presence of reduced nicotinamide adenine dinucleotide (NADH), endogenous NTR was detected in living cells, tissues and zebrafish. Cobalt chloride was used to induce chemical hypoxia to produce NTR, which generated enhanced fluorescence in cells and tumor tissues. Finally, two-photon fluorescence imaging of NTR was achieved in zebrafish at a penetration depth of up to 200 μm.
一种用于硝基还原酶的双光子荧光探针 FNTR 通过使用 9,9-二甲基-2-乙酰-芴-7-甲氨基（1）作为双光子荧光团和对硝基苄基碳酸酯基团作为硝基还原酶（NTR）的识别域来合成。探针和荧光团分别在单光子和双光子模式下进行了测试。与硝基还原酶反应后，FNTR 在 10 分钟内于 563nm 处的荧光增强了 130 倍，在 750nm 处检测到最大双光子作用截面值为 66GM。该探针具有较高的灵敏度，检测限低至 23.67ngml，具有较高的选择性、低细胞毒性和良好的光稳定性。在还原型烟酰胺腺嘌呤二核苷酸（NADH）存在下，可在活细胞、组织和斑马鱼中检测到内源性 NTR。氯化钴用于诱导化学缺氧以产生 NTR，从而在细胞和肿瘤组织中产生增强的荧光。最后，在斑马鱼中实现了 NTR 的双光子荧光成像，穿透深度可达 200μm。
