Drop coating deposition Raman spectroscopy of fluorescein isothiocyanate labeled protein.
Drop coating deposition Raman spectroscopy of fluorescein isothiocyanate labeled protein.
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DOI：
10.1366/000370210792973497
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发表时间：
2010-10
影响因子：
3.5
通讯作者：
Pechan T
中科院分区：
文献类型：
--
作者：
Zhang D;Vangala K;Jiang D;Zou S;Pechan T
以牛血清白蛋白(BSA)为模型蛋白，采用溶液和滴涂沉积拉曼(DCDR)采样技术首次系统研究了异硫氰酸荧光素(FITC)结合蛋白的正常拉曼光谱。FITC-BSA拉曼光谱主要由FITC拉曼特征主导，这些特征强烈地依赖于pH。目前，用10：1的FITC-BSA偶联物获得的dCDR检测灵敏度在总蛋白消耗量方面为45fmol，在激光探测体积下为~15attomol。与FITC-BSA溶液的拉曼光谱不同，FITC-BSA的拉曼特征是光稳定性的，而在FITC-BSA的dCDR光谱中观察到同时的FITC荧光和拉曼光漂白。FITC拉曼光漂白遵循与FITC标记率无关的时间常数的单指数衰减函数，而荧光背景光漂白要复杂得多，它强烈依赖于FITC标记率和样品条件。从机理上讲，FITC拉曼光漂白被认为是由于FITC分子在电子激发态下的光化学反应。FITC荧光光漂白包括浓度猝灭和光化学猝灭，后者可能涉及一种荧光不活跃但拉曼活性的光化学中间体。
Using bovine serum albumin (BSA) as the model protein normal Raman spectra of Fluorescein isothiocyanate (FITC) -conjugated protein was systematically studied for the first time using both solution and the drop coating deposition Raman (DCDR) sampling techniques. The FITC-BSA Raman spectra are dominated by the FITC Raman features that are strongly pH dependent. Current DCDR detection sensitivity obtained with a 10:1 FITC-BSA conjugate is 45 fmol in terms of total protein consumption and ~15 attomol at laser probed volume. Unlike the FITC-BSA solution Raman spectra where the FITC Raman features are photostable, concurrent FITC fluorescence and Raman photobleaching is observed in the DCDR spectra of FITC-BSA. While the FITC Raman photobleaching follows a single exponential decay function with a time constant independent of the FITC labeling ratio, the fluorescence background photobleaching is much more complicated and it depends strongly on the FITC labeling ratio and sample conditions. Mechanistically, the FITC Raman photobleaching is believed to be due to photochemical reaction of the FITC molecules in the electronically excited state. The FITC fluorescence photobleaching involves both concentration quenching and photochemical quenching, and the latter may involve a photochemical intermediate that is fluorescence inactive but Raman active.
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影响因子：
7.4
作者：
Campbell, Robert E.
通讯作者：
Campbell, Robert E.
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影响因子：
7.4
作者：
Han, Xiao X.;Cai, Lin J.;Ozaki, Yukihiro
通讯作者：
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影响因子：
3.3
作者：
Andreeva, Atanaska;Abarova, Silvia;Velitchkova, Maya
通讯作者：
Velitchkova, Maya
影响因子：
1.9
作者：
Boustany, NN
通讯作者：
Boustany, NN
影响因子：
3.9
作者：
Bui, Huong Thi;Umakoshi, Hiroshi;Kuboi, Ryoichi
通讯作者：
Kuboi, Ryoichi