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Enhanced light collection in fluorescence microscopy using self-assembled micro-reflectors

机译:使用自组装微反射镜增强荧光显微镜中的光收集

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摘要

In fluorescence microscopy, the signal-to-noise ratio (SNR) of the optical system is directly linked to the numerical aperture (NA) of the microscope objective, which creates detection challenges for low-NA, wide-field and high-throughput imaging systems. Here we demonstrate a method to increase the light collection efficiency from micron-scale fluorescent objects using self-assembled vapor-condensed polyethylene glycol droplets, which act as micro-reflectors for fluorescent light. Around each fluorescent particle, a liquid meniscus is formed that increases the excitation efficiency and redirects part of the laterally-emitted fluorescent light towards the detector due to internal reflections at the liquid-air interface of the meniscus. The three-dimensional shape of this micro-reflector can be tuned as a function of time, vapor temperature, and substrate contact angle, providing us optimized SNR performance for fluorescent detection. Based on these self-assembled micro-reflectors, we experimentally demonstrate ~2.5-3 fold enhancement of the fluorescent signal from 2-10 >μm sized particles. A theoretical explanation of the formation rate and shapes of these micro-reflectors is presented, along with a ray tracing model of their optical performance. This method can be used as a sample preparation technique for consumer electronics-based microscopy and sensing tools, thus increasing the sensitivity of low-NA systems that image fluorescent micro-objects.
机译:在荧光显微镜中,光学系统的信噪比(SNR)与显微镜物镜的数值孔径(NA)直接相关,这给低NA,宽视场和高通量成像带来了挑战系统。在这里,我们演示了一种方法,该方法使用自组装的蒸气冷凝聚乙二醇液滴(可充当荧光的微反射器)来提高微米级荧光对象的光收集效率。在每个荧光粒子周围形成液体弯液面,由于弯液面在液-气界面处的内部反射,液体弯液面提高了激发效率,并使部分侧向发射的荧光重定向到检测器。该微反射器的三维形状可以根据时间,蒸气温度和基板接触角进行调整,从而为荧光检测提供了优化的SNR性能。基于这些自组装的微反射器,我们实验证明2-10 >μ m大小的颗粒的荧光信号增强了〜2.5-3倍。给出了这些微反射镜的形成速率和形状的理论解释,以及它们的光学性能的射线追踪模型。此方法可用作基于消费类电子产品的显微镜和传感工具的样品制备技术,从而提高了对荧光微对象成像的低数值孔径系统的灵敏度。

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