Citation: LI Tian-Rui, HU Wei, LIU Zhi-Hong, LI Zhen. A Novel Two-photon Fluorescent Probe for Sensitive Determination of Nitroreductase and Live Cell Imaging[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(10): 1652-1659. doi: 10.11895/j.issn.0253-3820.181315
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A novel two-photon fluorescent probe, NNTR, was synthesized by using 6-methylamino-2-acetyl-naphthalen as a two-photon fluorophore and 4-nitrobenzyl chloridocarbonate as a recognition domain for nitroreductase (NTR). The spectroscopic characteristics and fluorescent response towards NTR were investigated using one-and two-photon modes, respectively. In the presence of NADH, NNTR could react with nitroreductase completely within 5 min. NNTR showed a 350-fold fluorescence enhancement at 510 nm in one-photo model and a 500-fold fluorescence enhancement at 810 nm in two-photo model. The maximal two-photon action cross-section value was calculated to be 66 GM. NNTR presented quick response, high sensitivity and good photostability towards NTR with a detection limit of 22 ng/mL. Cell survival rate of HeLa cells incubating with NNTR probes with different concentrations was investigated. Two-photon microscopic imaging nitroreductase in HeLa Cells treated with 5 μmol/L NNTR was successfully performed.
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