Low-cost optical lifetime assisted ratiometric glutamine sensor based on glutamine binding protein.
Low-cost optical lifetime assisted ratiometric glutamine sensor based on glutamine binding protein.
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DOI：
10.1016/j.ab.2008.08.018
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发表时间：
2008-12-01
影响因子：
2.9
通讯作者：
Tolosa, Leah
中科院分区：
文献类型：
--
作者：
Lam, Hung;Kostov, Yordan;Rao, Govind;Tolosa, Leah
本文报道了一种基于谷氨酰胺结合蛋白（QBP）的亚微摩尔谷氨酰胺无试剂荧光传感技术。S179 C突变体分别在-SH和N-末端位置用短寿命的丙烯酰二氢吡喃（寿命<5 ns）和长寿命的三（二苯甲酰甲烷）单（5-氨基-1，10-菲咯啉）铕（III）（寿命>300µs）标记。在谷氨酰胺存在下，丙烯酰胺的荧光被猝灭，而铕络合物的荧光保持恒定。在这份报告中，我们描述了一种创新的技术，所谓的寿命辅助比率传感，以区分两个荧光信号，使用最小的光学和功率要求。该方法利用两种荧光团的荧光寿命之间的巨大差异，通过在300 Hz和10 kHz之间交替激发光的调制频率来将各个荧光彼此隔离。结果是比率光学方法，其不需要用于每种染料的昂贵且高衰减的带通滤波器，而仅需要用于两者的一个长通滤波器。因此，提高了信噪比，同时简化了光学设置。最终产品是一种简单的传感设备，适用于低成本应用，如即时诊断或现场分析。
Here we report a reagentless fluorescence sensing technique for glutamine in the submicromolar range based on the glutamine binding protein (QBP). The S179C mutant is labeled with the short-lived acrylodan (lifetime<5ns) and the long-lived tris(dibenzoylmethane) mono(5-amino-1,10-phenanthroline)europium(III) (lifetime>300µs) at the –SH and the N-terminal positions, respectively. In the presence of glutamine the fluorescence of acrylodan is quenched, while the fluorescence of europium complex remains constant. In this report we describe an innovative technique, the so called lifetime assisted ratiometric sensing to discriminate the two fluorescence signals using minimal optics and power requirements. This method exploits the large difference between the fluorescence lifetimes of the two fluorophores to isolate the individual fluorescence from each other by alternating the modulation frequency of the excitation light between 300 Hz and 10 kHz. The result is a ratiometric optical method that does not require expensive and highly attenuating band pass filters for each of the dyes, but only one long pass filter for both. Thus, the signal to noise ratio is enhanced, and at the same time, the optical setup is simplified. The end product is a simple sensing device suitable for low-cost applications such as point-of-care diagnostics or in-the-field analysis.
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影响因子：
3.8
作者：
Larson, TM;Gawlitzek, M;Cacia, J
通讯作者：
Cacia, J
影响因子：
2.9
作者：
Mayer, C;Frauer, A;Pittner, F
通讯作者：
Pittner, F
DOI：
10.1152/ajpcell.2001.280.4.c867
发表时间：
2001-04-01
影响因子：
5.5
作者：
Dumaswala, UJ;Zhuo, L;Jacobsen, DW
通讯作者：
Jacobsen, DW
影响因子：
2.9
作者：
Bartolome, A;Bardliving, C;Tolosa, L
通讯作者：
Tolosa, L
影响因子：
7.4
作者：
Madaras, MB;Spokane, RB;Woodward, JR
通讯作者：
Woodward, JR