Abstract：
Single-molecule multiplexed detection is a high-promise toolkit for the expanding field of biosensing and molecular diagnostics. Among many single-molecule techniques available today for biomarker sensing including fluorescence, force, electrochemical, spectroscopic, barcoding, and other techniques, fluorescence-based approaches are arguably the most widely used methods due to their high sensitivity, selectivity, and readily available fluorophore-labeling schemes for a wide variety of biomolecules. However, multiplexed imaging using fluorescence techniques has proven to be challenging due to the sophisticated labeling schemes often requiring multiple FRET (fluorescence resonance energy transfer) pairs and/or excitation sources, which lead to overlapping signals and complicate data analysis. Here, we describe a single-molecule FRET method that enables multiplexed analysis while still using only one FRET pair, and thus the described approach is a significant step forward from conventional FRET methods.
摘要：
单分子多重检测是用于扩展生物传感和分子诊断领域的高前景工具包。在当今可用于生物标志物传感的许多单分子技术中，包括荧光，力,电化学,光谱学,条形码,和其他技术，基于荧光的方法可以说是最广泛使用的方法，由于它们的高灵敏度，选择性,以及用于多种生物分子的容易获得的荧光团标记方案。然而,使用荧光技术的多路成像已被证明是具有挑战性的，由于复杂的标记方案通常需要多个FRET（荧光共振能量转移）对和/或激发源，这导致重叠的信号和复杂的数据分析。这里,我们描述了一种单分子FRET方法，该方法能够进行多重分析，同时仍然仅使用一个FRET对，因此，所描述的方法是从传统的FRET方法向前迈出的重要一步。
