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基于高数值孔径照明的分辨率增强傅里叶叠层显微镜术

Resolution-enhanced Fourier ptychographic microscopy based on high-numerical-aperture illuminations.

作者信息

Sun Jiasong, Zuo Chao, Zhang Liang, Chen Qian

机构信息

Smart Computational Imaging (SCI) Laboratory, Nanjing University of Science and Technology, Nanjing, Jiangsu Province, 210094, China.

Jiangsu Key Laboratory of Spectral Imaging & Intelligent Sense, Nanjing University of Science and Technology, Nanjing, Jiangsu Province, 210094, China.

出版信息

Sci Rep. 2017 Apr 26;7(1):1187. doi: 10.1038/s41598-017-01346-7.

Abstract

High-resolution and wide field-of-view (FOV) microscopic imaging plays a central role in diverse applications such as high-throughput screening and digital pathology. However, conventional microscopes face inherent trade-offs between the spatial resolution and FOV, which are fundamental limited by the space-bandwidth product (SBP) of the optical system. The resolution-FOV tradeoff can be effectively decoupled in Fourier ptychography microscopy (FPM), however, to date, the effective imaging NA achievable with a typical FPM system is still limited to the range of 0.4-0.7. Herein, we report, for the first time, a high-NA illumination based resolution-enhanced FPM (REFPM) platform, in which a LED-array-based digital oil-immersion condenser is used to create high-angle programmable plane-wave illuminations, endowing a 10×, 0.4 NA objective lens with final effective imaging performance of 1.6 NA. With REFPM, we present the highest-resolution results with a unprecedented half-pitch resolution of 154 nm at a wavelength of 435 nm across a wide FOV of 2.34 mm, corresponding to an SBP of 98.5 megapixels (~50 times higher than that of the conventional incoherent microscope with the same resolution). Our work provides an important step of FPM towards high-resolution large-NA imaging applications, generating comparable resolution performance but significantly broadening the FOV of conventional oil-immersion microscopes.

摘要

高分辨率和宽视场(FOV)显微成像在高通量筛选和数字病理学等多种应用中发挥着核心作用。然而,传统显微镜在空间分辨率和视场之间面临固有的权衡,这在根本上受到光学系统的空间带宽积(SBP)的限制。不过,在傅里叶叠层显微术(FPM)中,分辨率-视场的权衡可以有效解耦。然而,迄今为止,典型FPM系统可实现的有效成像数值孔径(NA)仍限于0.4 - 0.7的范围。在此,我们首次报道了一种基于高NA照明的分辨率增强型FPM(REFPM)平台,其中基于LED阵列的数字油浸聚光镜用于创建高角度可编程平面波照明,赋予一个10×、0.4 NA的物镜最终1.6 NA的有效成像性能。利用REFPM,我们在2.34 mm的宽视场范围内,于435 nm波长下呈现了前所未有的154 nm半间距分辨率的最高分辨率结果,对应9850万像素的SBP(~比具有相同分辨率的传统非相干显微镜高50倍)。我们的工作为FPM迈向高分辨率大NA成像应用迈出了重要一步,产生了可比的分辨率性能,但显著拓宽了传统油浸显微镜的视场。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c65/5430655/a17249d65787/41598_2017_1346_Fig1_HTML.jpg

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