Active wide-field illumination for high-throughput fluorescence lifetime imaging.
Active wide-field illumination for high-throughput fluorescence lifetime imaging.
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DOI：
10.1364/ol.38.003976
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发表时间：
2013-10-01
期刊：
影响因子：
3.6
通讯作者：
Intes X
中科院分区：
文献类型：
--
作者：
Zhao L;Abe K;Barroso M;Intes X
宽视场荧光寿命成像允许大样本区域的快速成像，代价是对弱荧光信号的低灵敏度。为了克服这一挑战，我们开发了一种主动宽场照明（AWFI）策略来优化撞击空间强度，以获得整个样品的最佳荧光信号。我们证明了AWFI能够从浓度范围超过两个数量级的多孔板样品中准确估计寿命。我们进一步报道了其成功应用于定量Förster共振能量转移寿命细胞为基础的分析。总的来说，这种方法可以提高基于寿命的成像的准确性，在高采集速度下对具有大荧光强度分布的样品进行成像。
Wide-field fluorescence lifetime imaging allows for fast imaging of large sample areas at the cost of low sensitivity to weak fluorescence signals. To overcome this challenge, we developed an active wide-field illumination (AWFI) strategy to optimize the impinging spatial intensity for acquiring optimal fluorescence signals over the whole sample. We demonstrated the ability of AWFI to accurately estimate lifetimes from a multiwell plate sample with concentrations ranging over two orders of magnitude. We further reported its successful application to a quantitative Förster resonance energy transfer lifetime cell-based assay. Overall, this method allows for enhanced accuracy in lifetime-based imaging at high acquisition speed over samples with large fluorescence intensity distributions.