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上转换非线性结构光照明显微镜。

Upconversion Nonlinear Structured Illumination Microscopy.

机构信息

Institute for Biomedical Materials & Devices (IBMD), Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, Australia.

School of Mathematical and Physical Sciences, Faculty of Science, University of Technology Sydney, Ultimo, Sydney, NSW 2007, Australia.

出版信息

Nano Lett. 2020 Jul 8;20(7):4775-4781. doi: 10.1021/acs.nanolett.0c00448. Epub 2020 Mar 27.

Abstract

Video-rate super-resolution imaging through biological tissue can visualize and track biomolecule interplays and transportations inside cellular organisms. Structured illumination microscopy allows for wide-field super resolution observation of biological samples but is limited by the strong extinction of light by biological tissues, which restricts the imaging depth and degrades its imaging resolution. Here we report a photon upconversion scheme using lanthanide-doped nanoparticles for wide-field super-resolution imaging through the biological transparent window, featured by near-infrared and low-irradiance nonlinear structured illumination. We demonstrate that the 976 nm excitation and 800 nm upconverted emission can mitigate the aberration. We found that the nonlinear response of upconversion emissions from single nanoparticles can effectively generate the required high spatial frequency components in the Fourier domain. These strategies lead to a new modality in microscopy with a resolution below 131 nm, 1/7th of the excitation wavelength, and an imaging rate of 1 Hz.

摘要

通过生物组织进行视频速率超分辨率成像是可视化和跟踪细胞内生物分子相互作用和运输的关键手段。结构光照明显微镜允许对生物样本进行宽场超分辨率观察,但受到生物组织强烈光吸收的限制,这限制了成像深度并降低了成像分辨率。在这里,我们报告了一种基于镧系掺杂纳米粒子的上转换方案,用于通过生物透明窗口进行宽场超分辨率成像,其特点是近红外和低辐照度非线性结构光照。我们证明了 976nm 的激发和 800nm 的上转换发射可以减轻像差。我们发现,单个纳米粒子的上转换发射的非线性响应可以有效地在傅立叶域中产生所需的高空间频率分量。这些策略为显微镜带来了一种新的模式,分辨率低于 131nm,为激发波长的 1/7,成像速度为 1Hz。

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