Testing fluorescence lifetime standards using two-photon excitation and time-domain instrumentation: rhodamine B, coumarin 6 and lucifer yellow.
Testing fluorescence lifetime standards using two-photon excitation and time-domain instrumentation: rhodamine B, coumarin 6 and lucifer yellow.
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DOI：
10.1007/s10895-014-1368-1
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发表时间：
2014-07
影响因子：
2.7
通讯作者：
Frette, Oyvind
中科院分区：
文献类型：
--
作者：
Kristoffersen, Arne S.;Erga, Svein R.;Hamre, Borge;Frette, Oyvind
掌握有关荧光寿命标准的良好信息对于任何进行寿命实验的人来说都是至关重要的。在荧光光谱中使用寿命标准通常被认为是一个简单的过程，然而，许多早期的报告在寿命浓度依赖性、溶剂和其他技术方面受到限制。我们研究了荧光染料罗丹明B、香豆素6和荧光黄作为寿命标准的适宜性，特别是在时间域中用于双光子激发测量。我们测量了荧光团的吸收光谱和发射光谱，以确定我们应该使用哪些波长作为激发和合适的探测器范围。我们还测量了不同浓度(10−2-10−6 M)在水、乙醇和甲醇溶液中的寿命。我们观察到罗丹明B的寿命强烈地依赖于浓度。香豆素6提供了最稳定的寿命，对浓度和溶剂的依赖性可以忽略不计。研究还发现，荧光黄的寿命几乎不受浓度的影响。最后，我们发现，两种荧光团(罗丹明B/香豆素6，罗丹明B/路西法黄，香豆素6/路西法黄)的混合都产生了非常相似的双指数衰减寿命，与由单指数衰减测量的单独寿命非常相似。所有的寿命测量都是使用双光子激发进行的，并使用时间相关的单光子计数获得了时间域中的寿命数据。
Having good information about fluorescence lifetime standards is essential for anyone performing lifetime experiments. Using lifetime standards in fluorescence spectroscopy is often regarded as a straightforward process, however, many earlier reports are limited in terms of lifetime concentration dependency, solvents and other technical aspects. We have investigated the suitability of the fluorescent dyes rhodamine B, coumarin 6, and lucifer yellow as lifetime standards, especially to be used with two-photon excitation measurements in the time-domain. We measured absorption and emission spectra for the fluorophores to determine which wavelengths we should use for the excitation and an appropriate detector range. We also measured lifetimes for different concentrations, ranging from 10−2– 10−6 M, in both water, ethanol and methanol solutions. We observed that rhodamine B lifetimes depend strongly on concentration. Coumarin 6 provided the most stable lifetimes, with a negligible dependency on concentration and solvent. Lucifer yellow lifetimes were also found to depend little with concentration. Finally, we found that a mix of two fluorophores (rhodamine B/coumarin 6, rhodamine B/lucifer yellow, and coumarin 6/lucifer yellow) all yielded very similar lifetimes from a double-exponential decay as the separate lifetimes measured from a single-exponential decay. All lifetime measurements were made using two-photon excitation and obtaining lifetime data in the time-domain using time-correlated single-photon counting.
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影响因子：
1.9
作者：
Cerussi, AE;Maier, JS;Gratton, E
通讯作者：
Gratton, E
影响因子：
3.3
作者：
Magde, D;Rojas, GE;Seybold, PG
通讯作者：
Seybold, PG
影响因子：
3.5
作者：
de Grauw, CJ;Gerritsen, HC
通讯作者：
Gerritsen, HC
影响因子：
--
作者：
KEMNITZ, K;TAMAI, N;YOSHIHARA, K
通讯作者：
YOSHIHARA, K
影响因子：
2.3
作者：
PENZKOFER, A;LU, Y
通讯作者：
LU, Y