Pal Robert, Barker Abigail C J, Hummel Daniel, Pålsson Lars-Olof
Department of Chemistry, Durham University, Lower Mountjoy, Stockton Road, Durham, DH1 3LE, UK.
J Fluoresc. 2019 Jan;29(1):255-263. doi: 10.1007/s10895-018-2335-z. Epub 2018 Dec 26.
In this work we demonstrate that time domain techniques can be used successfully to monitor realtively weak modulations of the fluorescence in sensing applications. The metal sensing complex Newport Green DCF™ can detect selected transition metals in vivo as well as in vitro. Incremental addition of Ni and/or Zn (in vitro) lead to a substantial reduction in the yield of the fast component in a bi-exponential fluorescence decay (τ = 150-250 ps) from 60% to 30-35%. This is rationalised as an inhibition of intra-molecular electron transfer in the NPG sensing complex due to metal complexation. In order to explore this effect in cellulo, NIH 3 T3 mouse skin fibroplast cells were pre-incubated with set levels of Ni and Zn, at a constant concentration of NPG. The fluorescence modulation in cellullo was subsequently studied employing both time-resolved fluorescence microscopy and confocal fluorescence microscopy. In correlation with the in vitro observations, similar effects were observed on the fluorescence decay in cellulo.
在本研究中，我们证明了时域技术可成功用于监测传感应用中荧光的相对微弱调制。金属传感复合物纽波特绿DCF™ 能够在体内和体外检测特定的过渡金属。体外逐步添加镍和/或锌会导致双指数荧光衰减（τ = 150 - 250皮秒）中快速成分的产率从60%大幅降至30 - 35%。这被解释为由于金属络合抑制了NPG传感复合物中的分子内电子转移。为了在细胞内探究这种效应，将NIH 3T3小鼠皮肤成纤维细胞在固定浓度的NPG下，用设定水平的镍和锌进行预孵育。随后采用时间分辨荧光显微镜和共聚焦荧光显微镜研究细胞内的荧光调制。与体外观察结果相关，在细胞内的荧光衰减中也观察到了类似的效应。
