Method to identify and minimize artifacts induced by fluorescent impurities in single-molecule localization microscopy.
Method to identify and minimize artifacts induced by fluorescent impurities in single-molecule localization microscopy.
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DOI：
10.1117/1.jbo.23.10.106501
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发表时间：
2018-10
影响因子：
3.5
通讯作者：
Zhang HF
中科院分区：
文献类型：
--
作者：
Davis JL;Dong B;Sun C;Zhang HF
荧光杂质的存在一直是单分子成像中的一个障碍，它会导致样品的错误识别和更高的定位不确定性。光谱单分子定位显微镜可以记录每个随机单分子发射事件的完整荧光光谱。这种能力使我们能够量化样品制备步骤引入的荧光杂质的空间和光谱特性，基于此，我们开发了一种方法，可以有效地将荧光杂质与目标分子分离。
The existence of fluorescent impurities has been a long-standing obstacle in single-molecule imaging, which results in sample misidentification and higher localization uncertainty. Spectroscopic single-molecule localization microscopy can record the full fluorescent spectrum of every stochastic single-molecule emission event. This capability allows us to quantify the spatial and spectral characteristics of fluorescent impurities introduced by sample preparation steps, based on which we developed a method to effectively separate fluorescent impurities from target molecules.