同步多视图捕获与融合可提高宽场显微镜和光片显微镜的空间分辨率。

Simultaneous multiview capture and fusion improves spatial resolution in wide-field and light-sheet microscopy.

作者信息

Wu Yicong, Chandris Panagiotis, Winter Peter W, Kim Edward Y, Jaumouillé Valentin, Kumar Abhishek, Guo Min, Leung Jacqueline M, Smith Corey, Rey-Suarez Ivan, Liu Huafeng, Waterman Clare M, Ramamurthi Kumaran S, La Riviere Patrick J, Shroff Hari

机构信息

Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA.

Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

出版信息

Optica. 2016 Aug 20;3(8):897-910. doi: 10.1364/OPTICA.3.000897. Epub 2016 Aug 11.

Abstract

Most fluorescence microscopes are inefficient, collecting only a small fraction of the emitted light at any instant. Besides wasting valuable signal, this inefficiency also reduces spatial resolution and causes imaging volumes to exhibit significant resolution anisotropy. We describe microscopic and computational techniques that address these problems by simultaneously capturing and subsequently fusing and deconvolving multiple specimen views. Unlike previous methods that serially capture multiple views, our approach improves spatial resolution without introducing any additional illumination dose or compromising temporal resolution relative to conventional imaging. When applying our methods to single-view wide-field or dual-view light-sheet microscopy, we achieve a twofold improvement in volumetric resolution (~235 nm × 235 nm × 340 nm) as demonstrated on a variety of samples including microtubules in , SpoVM in sporulating , and multiple protein distributions and organelles in eukaryotic cells. In every case, spatial resolution is improved with no drawback by harnessing previously unused fluorescence.

摘要

大多数荧光显微镜效率低下,在任何时刻只能收集一小部分发射光。这种低效率不仅浪费了宝贵的信号,还降低了空间分辨率,并导致成像体积呈现出显著的分辨率各向异性。我们描述了通过同时捕获、随后融合和解卷积多个样本视图来解决这些问题的微观和计算技术。与之前串行捕获多个视图的方法不同,我们的方法在不引入任何额外照明剂量或不影响相对于传统成像时间分辨率的情况下提高了空间分辨率。当将我们的方法应用于单视图宽场或双视图光片显微镜时,我们在体积分辨率(约235纳米×235纳米×340纳米)上实现了两倍的提升,这在包括 中的微管、孢子形成中的SpoVM以及真核细胞中的多种蛋白质分布和细胞器等各种样本上得到了证明。在每种情况下,通过利用之前未使用的荧光,空间分辨率都得到了提高且没有缺点。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索