Chip-based label-free incoherent super-resolution optical microscopy.

IF 23.4 Q1 OPTICS
Nikhil Jayakumar,Luis E Villegas-Hernández,Weisong Zhao,Hong Mao,Firehun T Dullo,Jean-Claude Tinguely,Krizia Sagini,Alicia Llorente,Balpreet Singh Ahluwalia
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Abstract

The photo-kinetics of fluorescent molecules have enabled the circumvention of the far-field optical diffraction limit. Despite its enormous potential, the necessity to label the sample may adversely influence the delicate biology under investigation. Thus, continued development efforts are needed to surpass the far-field label-free diffraction barrier. The statistical similarity or finite coherence of the scattered light off the sample in label-free mode hinders the application of existing super-resolution methods based on incoherent fluorescence imaging. In this article, we present physics and propose a methodology to circumvent this challenge by exploiting the photoluminescence (PL) of silicon nitride waveguides for near-field illumination of unlabeled samples. The technique is abbreviated EPSLON, Evanescently decaying Photoluminescence Scattering enables Label-free Optical Nanoscopy. We demonstrate that such an illumination has properties that mimic the photo-kinetics of nano-sized fluorescent molecules, i.e., such an illumination permits incoherence between the scattered fields from various locations on the sample plane. Thus, the illumination scheme enables the development of a far-field label-free incoherent imaging system that is linear in intensity and stable over time, thereby permitting the application of techniques like structured illumination microscopy (SIM) and intensity-fluctuation-based optical nanoscopy (IFON) in label-free mode to circumvent the diffraction limit. In this proof-of-concept work, we observed a two-point resolution of ~ 180 nm on super-resolved nanobeads and resolution improvements between 1.9× to 2.8× over the diffraction limit, as quantified using Fourier Ring Correlation (FRC), on various biological samples. We believe EPSLON is a step forward within the field of incoherent far-field label-free super-resolution microscopy that holds a key to investigating biological systems in their natural state without the need for exogenous labels.
基于芯片的无标签非相干超分辨率光学显微镜。
荧光分子的光动力学使其能够绕过远场衍射极限。尽管其潜力巨大,但标记样品的必要性可能会对正在调查的微妙生物学产生不利影响。因此,需要持续的发展努力来超越远场无标签衍射势垒。样品在无标记模式下散射光的统计相似性或有限相干性阻碍了现有基于非相干荧光成像的超分辨率方法的应用。在本文中,我们介绍了物理学并提出了一种方法,通过利用氮化硅波导的光致发光(PL)对未标记样品进行近场照明来规避这一挑战。该技术缩写为EPSLON,即倏逝衰减光致发光散射技术,可实现无标签光学纳米显微镜。我们证明了这种照明具有模拟纳米级荧光分子的光动力学的特性,即,这种照明允许来自样品平面上不同位置的散射场之间的不相干。因此,该照明方案能够开发出一种远场无标签非相干成像系统,该系统强度呈线性且随时间稳定,从而允许在无标签模式下应用结构照明显微镜(SIM)和基于强度波动的光学纳米镜(IFON)等技术,以绕过衍射极限。在这项概念验证工作中,我们观察到超分辨率纳米球的两点分辨率为~ 180 nm,并且在衍射极限上分辨率提高了1.9 x到2.8 x,使用傅立叶环相关(FRC)对各种生物样品进行了量化。我们相信EPSLON是在非相干远场无标签超分辨率显微镜领域向前迈出的一步,它是研究生物系统在自然状态下不需要外源标签的关键。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Light-Science & Applications
Light-Science & Applications 数理科学, 物理学I, 光学, 凝聚态物性 II :电子结构、电学、磁学和光学性质, 无机非金属材料, 无机非金属类光电信息与功能材料, 工程与材料, 信息科学, 光学和光电子学, 光学和光电子材料, 非线性光学与量子光学
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803
审稿时长
2.1 months
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