Sun Jiasong, Zuo Chao, Zhang Liang, Chen Qian
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.
Sci Rep. 2017 Apr 26;7(1):1187. doi: 10.1038/s41598-017-01346-7.
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.
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。