Steps towards spatially resolved single molecule detection in solution

H P Mathis, G Kalusche, B Wagner, J S McCaskill
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Abstract

Spatially resolved single molecule detection in solution is a prerequisite for molecule tracking and provides new opportunities in single molecule manipulation, for example in a sample flow. A detector with fast timing properties and high spatial resolution is required for these purposes. We introduce a concept for spatially resolved optical single molecule detection in an epi-illuminated microscope using a novel kind of detector and a new algorithm in configurable hardware for intelligent data processing. The analysis is performed on a parallel hardware interface and includes burst detection. It can be extended to on-line spatial and temporal data analysis. This detector will be used in combination with a molecular sorter where molecules are sorted in a microstructured flow device made of silicon. In a first step towards such sorting in these microstructures, we show that a reliable detection of single molecules in silicon microstructures is possible in zero-dimensional detection volumes. The noise structure of the data is analysed in terms of Poissonian statistics and it is shown that in the smallest structure used (depth 20 μm) a signal-to-noise ratio of 40 is achieved.

在溶液中实现空间分辨单分子检测的步骤
溶液中的空间分辨单分子检测是分子跟踪的先决条件,并为单分子操作提供了新的机会,例如在样品流中。为此,需要一种具有快速定时特性和高空间分辨率的探测器。我们提出了一种利用新型检测器和可配置硬件进行智能数据处理的新算法在外延发光显微镜中进行空间分辨光学单分子检测的概念。分析是在并行硬件接口上进行的,包括突发检测。它可以扩展到在线时空数据分析。该探测器将与分子分选器结合使用,分子在硅制成的微结构流动装置中进行分选。在这些微结构中进行这种分选的第一步,我们表明可以在零维检测体积中可靠地检测硅微结构中的单分子。根据泊松统计分析了数据的噪声结构,结果表明,在最小的结构(深度为20 μm)中,信噪比达到40。
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