Fluorescence lifetime biosensing with DNA microarrays and a CMOS-SPAD imager.
Fluorescence lifetime biosensing with DNA microarrays and a CMOS-SPAD imager.
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DOI：
10.1364/boe.1.001302
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发表时间：
2010-11-04
影响因子：
3.4
通讯作者：
Arlt J
中科院分区：
文献类型：
--
作者：
Giraud G;Schulze H;Li DU;Bachmann TT;Crain J;Tyndall D;Richardson J;Walker R;Stoppa D;Charbon E;Henderson R;Arlt J
染料分子的荧光寿命是局部微环境的敏感报告因子，通常与荧光团浓度无关，可用作区分具有光谱重叠发射的分子的手段。因此，它是生物传感中潜在的强大的多重检测方式，但需要生物分析物浓度典型的极低光水平操作、长数据采集周期和片上处理能力才能实现这些优势。我们在此报告使用 CMOS-SPAD 成像仪结合 DNA 微阵列和 TIRF 激发几何结构获得的荧光寿命数据。这使得能够在不到 26 秒的曝光时间内获取 320 像素 FLIM 图的单光子到达时间直方图。由此，我们在低至 10 nM 的浓度下解析了与染料标记的 HCV 和量子点标记的 HCMV 核酸靶标相对应的不同寿命特征。
Fluorescence lifetime of dye molecules is a sensitive reporter on local microenvironment which is generally independent of fluorophores concentration and can be used as a means of discrimination between molecules with spectrally overlapping emission. It is therefore a potentially powerful multiplexed detection modality in biosensing but requires extremely low light level operation typical of biological analyte concentrations, long data acquisition periods and on-chip processing capability to realize these advantages. We report here fluorescence lifetime data obtained using a CMOS-SPAD imager in conjunction with DNA microarrays and TIRF excitation geometry. This enables acquisition of single photon arrival time histograms for a 320 pixel FLIM map within less than 26 seconds exposure time. From this, we resolve distinct lifetime signatures corresponding to dye-labelled HCV and quantum-dot-labelled HCMV nucleic acid targets at concentrations as low as 10 nM.