Enhanced DNA detection using a multiple pulse pumping scheme with time-gating (MPPTG).
Enhanced DNA detection using a multiple pulse pumping scheme with time-gating (MPPTG).
复制标题
使用具有时间选通 (MPPTG) 的多脉冲泵浦方案增强 DNA 检测。
DOI：
10.1039/c8an00136g
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Gryczynski,Zygmunt
中科院分区：
文献类型：
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作者：
Kimball,JosephD;Maliwal,Badri;Raut,SangramL;Doan,Hung;Nurekeyev,Zhangatay;Gryczynski,Ignacy;Gryczynski,Zygmunt
嵌入剂与DNA结合诱导的荧光信号增强已被广泛应用于各种DNA检测方法中。在大多数情况下，荧光强度的增加与荧光寿命的伴随增加相关。荧光寿命的增加提供了增加检测灵敏度的额外机会。在本文中，我们提出了一种新的方法，显着提高检测微量DNA浓度的灵敏度。该方法是基于同时使用的时间选通检测和多脉冲泵浦。通过使用短脉冲的校准突发，我们极大地增强了长寿命荧光物质的贡献，从而使容易的时间选通检测。使用经典的DNA嵌入剂-溴化亚铵（EtBr）-作为我们的新型多脉冲泵浦和时间门控检测技术的一个例子，我们能够仅用3个脉冲激发将检测灵敏度提高70倍以上。这种方法是通用的，并且可以与任何分析探针（表现出约10倍的寿命变化）一起使用，所述分析探针在与靶标结合后显示荧光信号和荧光寿命的增加。
Fluorescence signal enhancement induced by the binding of intercalators to DNA has been broadly utilized in various DNA detection methods. In most instances the increase in fluorescence intensity is associated with a concomitant increase of fluorescence lifetime. This increase of the fluorescence lifetime presents an additional opportunity to increase detection sensitivity. In this paper, we present a new approach to significantly enhance the sensitivity in detecting minute DNA concentrations. The approach is based on simultaneous use of time-gated detection and multi-pulse pumping. By using a calibrated burst of short pulses we greatly enhance the contribution of long-lived fluorescence species, thus enabling easy time-gated detection. Using a classic DNA intercalator – Ethidium Bromide (EtBr) – as an example with our novel multi-pulse pumping and time-gated detection technique, we were able to increase detection sensitivity over 70-fold with only 3 pulse excitation. This approach is generic and can be used with any analytical probe (exhibiting about 10 times change in lifetime) that shows an increase in fluorescence signal and fluorescence lifetime upon binding to a target.