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Single-molecule detection using confocal fluorescence detection: Assessment of optical probe volumes

机译:使用共聚焦荧光检测的单分子检测:光学探针体积的评估

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摘要

We have constructed a simple component-based confocal detection system, that is capable of fluorescence detection at the single-molecule level. The component-based format maximises flexibility and reduces start-up costs. A new model for the optical probe volume is proposed, that is based on the 1/e~(2) Gaussian intensity contour of a laser beam focused to the diffraction limit. Observation of the onset of single-molecule detection in our experimental system confirms that this model is more appropriate than a simple cylindrical approximation for a probe volume defined by a wide, tightly focused laser beam.
机译:我们构建了一个简单的基于组件的共聚焦检测系统,该系统能够在单分子水平上进行荧光检测。基于组件的格式最大程度地提高了灵活性,并降低了启动成本。提出了一种基于聚焦到衍射极限的激光束的1 / e〜(2)高斯强度轮廓的光学探头体积的新模型。在我们的实验系统中对单分子检测开始的观察证实,对于由宽且紧密聚焦的激光束定义的探针体积,该模型比简单的圆柱近似更合适。

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